王雍^,, 李思妍, 何思锐, 张迪, 连帅, 王建发, 武瑞^,黑龙江八一农垦大学动物科技学院/黑龙江省牛病防制重点实验室,黑龙江大庆 163319

Prediction and Bioinformatics Analysis of BLV-miRNA Transboundary Regulation of Human Target Genes

WANG Yong^,, LI SiYan, HE SiRui, ZHANG Di, LIAN Shuai, WANG JianFa, WU Rui^,College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University/Heilongjiang Provincial Key Laboratory of Prevention and Control of Bovine Diseases, Daqing 163319, Heilongjiang

通讯作者: 武瑞,Tel:13836961026;E-mail: fuhewu@126.com

责任编辑: 林鉴非
收稿日期:2020-02-23接受日期:2020-07-29网络出版日期:2021-02-01

基金资助:

国家自然科学基金.2041340046 黑龙江省自然科学基金.YQ2019C014 研究生创新科研项目.YJSCX2019-Y39

Received:2020-02-23Accepted:2020-07-29Online:2021-02-01
作者简介 About authors
王雍,Tel:13251599676;E-mail: bywy0209@126.com。









摘要
【目的】评估牛白血病病毒（BLV）来源的miRNAs跨界调控人源基因的风险。对BLV-miRNA可能带来的食品安全问题及对人体健康可能造成何种影响进行前瞻性研究,为未来实际生产中地方流行性白血病防控措施执行的必要性研究奠定基础,对BLV与人类疾病间关联性的研究提供理论指导。【方法】首先使用mirbase网站对BLV miRNA的成熟序列进行查询,通过miRanda软件对BLV编码的10种miRNA（BLV-miR-B1-3P,5P、BLV-miR-B2-3P,5P、BLV-miR-B3-3P,5P、BLV-miR-B4-3P,5P、BLV-miR-B5-3P,5P）进行靶基因预测,并选取每个BLV-miRNA评分前10的候选靶基因（去除重复基因后共88个）进行功能分析,对受到多个BLV miRNA共同调控的候选靶基因使用RNAhybrid软件进行二次预测验证,并对其功能进行分析。【结果】 BLV编码的10种miRNA经预测后分别获得1 630—16 383个靶基因不等。对评分前十的共计88个候选靶基因进行功能分析后发现,其中18个基因无相关功能报道;36个候选靶基因与肿瘤性疾病的发生发展存在相关性。2个候选靶基因可以对细胞周期起调控作用;16个候选靶基因参与细胞信号转导的调控;14个候选靶基因在细胞结构/骨架蛋白的形成中发挥作用;细胞的增殖与凋亡的功能表现成拮抗关系,往往促进增殖的基因同时也可以抑制细胞凋亡,共有13个基因对细胞的增殖和凋亡起调节作用,有趣的是,这13的候选靶基因对细胞增殖凋亡功能的调节是双向性的,但不能明确BLV miRNA对细胞的调节到底是更趋向于增殖还是凋亡,因此仍需要后续研究深入探讨;2个候选靶基因对细胞分化起调节作用;16个候选靶基因对细胞的迁移/侵袭功能起调节作用,再次提示BLV miRNA与肿瘤性疾病可能存在更重要的关联性。7个候选靶基因可能在乳腺细胞的分化、迁移、侵袭过程中发挥重要作用,提示BLV与人乳腺癌相关性的研究中,可以从BLV miRNA的角度深入探讨;BLV-B4-3P的2个候选靶基因Ⅰ型胶原α1链基因（COL1A1）、断裂点簇集区（BCR）对人急性淋巴细胞白血病（ALL）具有调节作用。此外,可以被多个BLV miRNA共同靶向的候选靶基因均属于黏蛋白家族（MUC5B、MUC12和 MUC16）,且均可以在结肠中表达,对结肠黏膜的形成产生影响。【结论】外源性BLV miRNA可能跨界调控细胞周期、信号转导、结构/骨架、增殖、凋亡、分化、迁移/侵袭相关等细胞功能相关基因,破坏细胞结构;BLV miRNA与人乳腺癌的相关性可能表现在人乳腺癌细胞的分化、迁移、和侵袭过程中;而BLV-miR-B4-3p本身与白血病相关miR 29a共享同一种子区域,可能对人急性淋巴细胞白血病的发生发展造成影响;外源性BLV miRNA具有靶向抑制黏蛋白基因（MUC5B、MUC12、MUC16）表达,通过破坏肠黏膜形成这一途径,跨界调控人源基因的风险。
关键词： 生物信息学;牛白血病病毒;跨界调控;人源基因;miRNA

Abstract
【Objective】To assess risk of regulation of human-derived genes by miRNAs derived from bovine leukemia virus (BLV), the prospective research on the possible food safety problems and the possible impact on human health caused by BLV-miRNA were carried out, which would lay the foundation for the necessary research on the implementation of Enzootic Bovine Leukosis (EBL) prevention and control measures in actual production in the future, and provide theoretical guidance for the study of the relationship between BLV and human diseases.【Method】 In this study, the mature sequence of BLV miRNAs was first queried using mirbase website, and the miRanda software was used to predict target genes. The predictive 10 miRNAs (BLV-miR-B1-3P,5P, BLV-miR-B2-3P, 5P, BLV-miR-B3-3P,5P, BLV-miR-B4-3P,5P, and BLV-miR-B5-3P,5P) were encoded by BLV. The top 10 candidate target genes of each BLV-miRNA score were selected for functional analysis, including a total of 88 duplicated genes. The candidate target genes co-regulated by multiple BLV miRNAs were verified by secondary prediction using RNAhybrid software, and their functions were analyzed. 【Result】The ten miRNAs encoded by BLV were predicted to obtain 1 630-16 383 target genes, respectively. After functional analysis of eighty-eight candidate target genes in the top ten, it was found that eighteen of them had no relevant functional reports. Thirty-six candidate target genes were related to the occurrence and development of neoplastic diseases. Two candidate target genes could regulate cell cycle. Sixteen candidate target genes were involved in the regulation of cell signal transduction. Fourteen candidate target genes played a role in the formation of structure/cytoskeleton proteins. The function of cell proliferation and apoptosis showed an antagonistic relationship and the genes that often promoting proliferation could also suppress apoptosis. A total of thirteen genes played a regulatory role in cell proliferation and apoptosis. Interestingly, the regulation of the thirteen candidate target genes on cell proliferation and apoptosis was bidirectional. However, it was not clear whether the regulation of BLV miRNA towards cells was more prone to proliferation or apoptosis, so further studies were still needed to discuss in depth. Two candidate target genes could regulate cell differentiation. The sixteen candidate target genes played a role in regulating cell migration/invasion function, again suggesting that BLV miRNA might have a more important correlation with neoplastic diseases. The seven candidate target genes might play an important role in the differentiation, migration and invasion of breast cells, suggesting that the study on the correlation between BLV and human breast cancer could be further discussed from the perspective of BLV miRNA. Two candidate target genotypes of BLV-B4-3P, Collagen 1 chain gene (COL1A1), had a regulatory effect on human acute lymphoblastic leukemia (ALL). In addition, candidate target genes that could be co-targeted by multiple BLV miRNAs belong to the mucin family (MUC5B, MUC12 and MUC16), and be expressed in the colon, influencing the formation of colon mucosa.【Conclusion】Exogenous BLV miRNA might transboundary regulate cell cycle signal transduction structure/cytoskeleton proliferation apoptosis differentiation migration/invasion related cell function related genes and destroy cell structure. The correlation between BLV miRNA and human breast cancer might be shown in the process of differentiation, migration and invasion of human breast cancer cells. BLV-miR-B4-3p shared a seed sequence with miR 29a, which might affect the occurrence and development of human acute lymphoblastic leukemia. Exogenous BLV miRNA had the target of inhibiting the expression of mucin genes, such as MUC5B, MUC12, and MUC16, through the destruction of intestinal mucosa formation to achieve transboundary regulation of human gene risk.
Keywords：bioinformatics;bovine leukemia virus;transboundary regulation;human-derived genes;miRNA


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本文引用格式
王雍, 李思妍, 何思锐, 张迪, 连帅, 王建发, 武瑞. BLV-miRNA跨界调控人类靶基因预测及生物信息学分析[J]. 中国农业科学, 2021, 54(3): 662-674 doi:10.3864/j.issn.0578-1752.2021.03.019
WANG Yong, LI SiYan, HE SiRui, ZHANG Di, LIAN Shuai, WANG JianFa, WU Rui. Prediction and Bioinformatics Analysis of BLV-miRNA Transboundary Regulation of Human Target Genes[J]. Scientia Acricultura Sinica, 2021, 54(3): 662-674 doi:10.3864/j.issn.0578-1752.2021.03.019


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0 引言

【研究意义】牛白血病病毒（BLV）是牛群中常见的传染性病毒之一,在世界各地均有分布,阳性牛群会呈持续感染状态,通常为亚临床型,只有约5%的BLV阳性牛会表现出淋巴瘤的症状^[1]。最新的调查结果显示,BLV在我国牛群中的发病率约为49%^[2]。牛群中出现BLV感染后,会出现产奶量下降^[3]、平均寿命降低^[4]、免疫失败^[5]等诸多问题。基于BLV临床症状不明显的特点,在目前的实际生产中并不能引起饲养者的重视,而BLV编码的miRNA具有靶向调控人源基因的风险,可能会对乳制品食用者的健康产生威胁,因此对BLV miRNA的研究对乳制品安全风险的预防具有重要意义。【前人研究进展】乳汁中发现的BLV可能引发全球性的公共卫生问题,1974年,MCCLURE等用自然感染BLV奶牛乳汁（未巴氏消毒）饲喂6只初生黑猩猩,连续饲喂至第30周后全部黑猩猩出现了嗜睡、厌食、白细胞增多、贫血和进行性肺炎症状,第35周和第46周时有两只黑猩猩分别死亡,病死黑猩猩被确诊患有红细胞白血病（erythroleukemia）^[6]。1976年,GRAVES等发现BLV可以体外感染牛、羊、猫、蝙蝠、类人猿和人类细胞（人胚肺二倍体细胞）。2015年以来,美国、澳大利亚和阿根廷乳腺癌患者乳腺组织样品中相继被检出BLV核酸片段^[7]。2019年4月,BUEHRING等在美国加州当地招募了95名妇女志愿者,对其血液中BLV核酸和BLV IgA、IgM、IgG进行了检测,发现BLV核酸阳性数为33,IgG和IgA抗体阳性数均为30,IgM抗体阳性数为55^[8]。因此,虽然尚无充分证据表明BLV是导致人乳腺癌的元凶,但BLV引发的公共卫生问题愈发受到关注。BLV作为逆转录病毒,但已经有研究者通过高通量测序技术证实BLV可编码miRNA,且在BLV阳性牛的肉和乳汁中发现了BLV miRNAs的存在^[9,10]。机体内BLV主要依靠miRNA调控牛淋巴细胞功能,当BLV感染后,其核酸序列整合到B淋巴细胞基因组中,导致B细胞在宿主体内异常增殖^[11]。但是由于B细胞淋巴瘤中缺乏5′LTR驱动的RNA聚合酶II（PolII）,BLV在原代B细胞淋巴瘤中出现转录沉默,无法检测到病毒mRNA和蛋白质^[12]。基于此,研究者在对BLV致病机制的研究过程中发现,白血病牛B淋巴细胞中存在大量高度保守的由RNA聚合酶III（PolIII）转录的BLV编码的miRNA^[9],也打破了RNA病毒不编码miRNA以避免其基因组或mRNA的非生产性切割的传统认知。miRNA相比mRNA具有极高的稳定性,乳品中BLV-miRNA具有危害人类健康的潜在风险,BLV可编码5类共计10种miRNA,其中miR-B4-3p、miR-B2-5p、miR-B5-5p、miR-B1-3p、miR-B5-3p与牛白血病的发生相关^[13],这些miRNA主要靶向颗粒酶A（GZMA）、棕榈酰蛋白硫酯酶1（PPT1）、膜联蛋白A1（ANXA1）、促分裂原活化蛋白激酶激酶1（MAP2K1）、磷酸肌醇3激酶（PIK3CG）、FBJ鼠科骨肉瘤病毒原癌基因（FOS）等癌相关基因,清除上述miRNA后BLV不能诱导白血病发生^[14]。BLV与人类T淋巴细胞白血病病毒1型（HTLV-1）存在共同的致瘤机制和结构高度相似的致瘤miRNA^[15]。miRNA可以跨物种、跨代际稳定存在,并在不同物种间、代际间跨界调控基因表达,且miRNA跨界调控的普适性规律已得到广泛认可^[16]。BLV修饰后的牛树突状细胞外泌体携带有致瘤miRNA,并且牛奶外泌体所携miRNA可跨界调控人类免疫细胞功能^[17]。【本研究切入点】前人对BLV可能对人类健康产生风险的研究主要集中于BLV与人乳腺癌之间的联系,本文以miRNA跨界调控的普适性作为切入点, BLV作为一种可以编码miRNA的逆转录病毒,其所编码的miRNA可以在牛奶的外泌体中发现^[10],具有跨界调控人源基因的食品安全风险。【拟解决的关键问题】通过miRanda软件,预测了BLV miRNAs可能调控的人源靶基因,并对BLV miRNAs的候选靶基因功能进行查询并分析,对进一步认识BLV miRNAs跨界调控人源基因风险的研究具有重要的科学意义和实际价值。

1 材料与方法

1.1 信息采集

通过mirbase（www.mirbase.org）查询BLV-miR- B1-3P,5P、BLV-miR-B2-3P,5P、BLV-miR-B3-3P,5P、BLV-miR-B4-3P,5P、BLV-miR-B5-3P,5P成熟序列（表1）。

Table 1
表1
表1BLV miRNAs成熟序列
Table 1BLV miRNAs mature sequence

miRNA名称 miRNA name	成熟序列 Mature sequence
BLV-miR-B1-3P	UCAGUGUACCAUCACAAGCCUCU
BLV-miR-B1-5P	AGGCUGUGGUGGUGCACUGGCUU
BLV-miR-B2-3P	UGCGUGUCGCUCAGUCAUUUU
BLV-miR-B2-5P	AUGACUGAGUGUAGCGCAGAGA
BLV-miR-B3-3P	UAACGCUGACGGGGGCGAUUUCU
BLV-miR-B3-5P	AUCCCCCUGCCAGCGUUGGUC
BLV-miR-B4-3P	UAGCACCACAGUCUCUGCGCCUUU
BLV-miR-B4-5P	GCGGGAGGCUCUGGUGCUGG
BLV-miR-B5-3P	CUCGAGCCGCAACCUCCCUUUCU
BLV-miR-B5-5P	AGGAAGGUUGUGGCUCAGAGGU

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1.2 靶基因预测

miRanDa软件主要通过miRNA和mRNA间的序列互补匹配程度和形成复合结构的自由能两个因素来判断miRNA与候选靶基因的结合位点,根据序列匹配的打分值和对应的自由能预测结合位点的可能性,当分值大于150时,可以被认定为该miRNA 的候选靶基因,由于不依赖结合位点的保守性,可以更加广泛的对miRNA的结合位点进行评估,因此我们在哈尔滨医科大学李春权老师帮助下,于2019年7月在哈尔滨医科大学大庆分校区应用miRanda（V1.9）软件对BLV-miR-B1-3P,5P、BLV-miR-B2-3P,5P、BLV- miR-B3-3P,5P、BLV-miR-B4-3P,5P、BLV-miR-B5-3P,5P序列进行靶基因预测分析。

1.3 靶基因功能查询

通过NCBI、Bing检索工具检索每个BLV-miRNA所预测的靶基因评分前十位的靶基因功能。通过RNAhybrid^[18]对多个BLV-miRNA共同调控的靶基因进行二次预测验证,当最小自由能（△G）≤-15 kcal·mol^-1时结合的可能性较高。

2 结果

2.1 BLV miRNAs靶向人源基因预测

通过miRanda软件对BLV miRNAs进行靶基因预测,BLV miRNAs分别获得了1 630—16 383个靶基因不等（图1）。

图1

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图1BLV miRNAs靶基因预测

Fig. 1BLV miRNAs target gene prediction results

2.2 BLV miRNAs靶向评分前十位基因功能分析

对BLV-miRNAs的靶基因进行了分别预测,并对每个BLV-miRNA评分排名前十的靶基因（去除重复基因后共88个）功能进行了整理分析,由于18个候选靶基因没有相关功能报道,最终本研究对70个靶基因的功能进行了分析,其中有36个基因的功能与肿瘤的发生发展存在相关性。这些候选靶基因所表达的蛋白在细胞的周期、信号转导、结构/骨架、增殖、凋亡、分化、迁移/侵袭功能中发挥了重要作用（表2—7）。

Table 2
表2
表2参与调控细胞周期的候选靶基因
Table 2Candidate target genes involved in cell cycle regulation

序号 Number	基因名称 Gene name	miRNA	评分 Score	与细胞周期关系 Relationship with cell cycle
1	MGRN1	BLV-miR-B2-3p	308	MGRN1介导的α-微管蛋白在细胞间期起泛素化作用[19] MGRN1-mediated α-tubulin ubiquitination in the intercellular phase
2	CDC42	BLV-miR-B4-5p	925	调控细胞周期[20] Regulating of cell cycle

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Table 3
表3
表3参与细胞信号转导的候选靶基因
Table 3Candidate target genes involved in cell signal transduction

序号 Number	基因名称 Gene name	miRNA miRNA	评分 Score	与信号转导关系 Relationship with signal transduction
1	NOTCH3	BLV-miR-B1-3p	600	编码果蝇I型膜蛋白缺口的人类同源物,建立细胞间信号传导途径,其在神经发育中起关键作用 The human homologues encoding gaps in Drosophila type I membrane proteins, establishing intercellular signal transduction pathways, which play a key role in neural development
2	MUC12	BLV-miR-B1-5p	4260	编码膜糖蛋白,在细胞内信号传导中发挥作用/黏液素在上皮表面形成的保护性黏液屏障中起重要作用 It encodes membrane glycoproteins and plays a role in intracellular signaling, and mucin plays an important role in the protective mucus barrier formed by the epithelial surface
3	MUC21	BLV-miR-B1-5p	3738	膜结合糖蛋白,在上皮表面形成保护性黏膜屏障中起重要作用,也在细胞内信号传导中起作用 Membrane-bound glycoproteins play an important role in the formation of a protective mucosal barrier on the epithelial surface and in intracellular signaling
4	ATF3	BLV-miR-B2-5p	180	编码哺乳动物激活转录因子;参与细胞应激反应 Encode mammalian activation transcription factors and participate in cellular stress response
5	NR2C2	BLV-miR-B2-5p	179	该基因编码属于核激素受体家族的蛋白质,该家族的成员充当配体激活的转录因子 The gene encodes proteins belonging to the nuclear hormone receptor family, members of which act as ligand-activated transcription factors
6	OR14I1	BLV-miR-B2-5p	187	负责识别和G蛋白介导的气味信号转导 Responsible for recognition and G protein-mediated odor signal transduction
7	POU3F3	BLV-miR-B2-3p	444	该基因编码含有POU结构域的蛋白质,该蛋白质起着转录因子的作用 The gene encodes a protein containing a POU domain, which acts as a transcription factor
8	TIAL1-201	BLV-miR-B2-5p	180	该基因编码的蛋白质是RNA结合蛋白家族的成员,调节各种活动,包括翻译控制,剪接和凋亡 The gene encodes proteins that are members of the RNA-binding protein family and regulate various activities including translation control, splicing, and apoptosis
9	TXNL1-204	BLV-miR-B2-5p	175	TXNL1- XRCC1是一种新型的调控途径,氧化还原传感器TXNL1在液相内吞中起调节作用[21] TXNL1- XRCC1 is a new regulatory approach, and the redox sensor TXNL1 plays a regulatory role in liquid phase endocytosis
10	MUC12	BLV-miR-B3-3p	1893	编码膜糖蛋白,在细胞内信号传导中发挥作用/黏液素在上皮表面形成的保护性黏液屏障中起重要作用 It encodes membrane glycoproteins and plays a role in intracellular signaling, and mucin plays an important role in the protective mucus barrier formed by the epithelial surface
11	AHNAK2	BLV-miR-B3-5p	6265	编码的蛋白质可通过与钙通道蛋白结合而在钙信号传导中起作用 The encoded protein can play a role in calcium signaling by binding to calcium channel proteins
12	EPN1	BLV-miR-B4-5p	1506	促进囊泡的内吞作用[22] Promoting endocytosis of vesicles
13	TSPAN14	BLV-miR-B4-5p	1052	作为跨膜蛋白,可调节内质网出口[23] As a transmembrane protein, it can regulate the export of endoplasmic reticulum
14	MUC12	BLV-miR-B4-5p	1045	编码膜糖蛋白,在细胞内信号传导中发挥作用/黏液素在上皮表面形成的保护性黏液屏障中起重要作用 It encodes membrane glycoproteins and plays a role in intracellular signaling, and mucin plays an important role in the protective mucus barrier formed by the epithelial surface
15	MUC12	BLV-miR-B5-3p	506	编码膜糖蛋白,在细胞内信号传导中发挥作用/黏液素在上皮表面形成的保护性黏液屏障中起重要作用 It encodes membrane glycoproteins and plays a role in intracellular signaling, and mucin plays an important role in the protective mucus barrier formed by the epithelial surface
16	MUC12	BLV-miR-B5-5p	3373	编码膜糖蛋白,在细胞内信号传导中发挥作用/黏液素在上皮表面形成的保护性黏液屏障中起重要作用 It encodes membrane glycoproteins and plays a role in intracellular signaling, and mucin plays an important role in the protective mucus barrier formed by the epithelial surface

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Table 4
表4
表4参与合成细胞结构/骨架蛋白的候选靶基因
Table 4Candidate target genes involved in the synthesis of cell structure / skeleton protein

序号 Number	基因名称 Gene name	miRNA miRNA	评分 Score	与结构/骨架关系 Relationship with structure / skeleton
1	MYO19	BLV-miR-B1-3p	596	肌球蛋白,为肌肉收缩,胞质分裂和细胞器运输等过程提供动力[24] It is myosin that powers muscle contraction, cytokinesis and organelle transport
2	BSN	BLV-miR-B1-5p	2197	编码的tau蛋白是神经细胞的骨架成分 The encoded tau protein is a skeleton component of nerve cells
3	KLC1	BLV-miR-B1-5p	2308	与肌动蛋白重链结合、参与了囊泡、线粒体和高尔基体等物质的结合 It binds to actin heavy chains and is involved in the binding of vesicles, mitochondria and golgi bodies
4	OBSCN	BLV-miR-B1-5p	2921	该基因编码的Obscurins蛋白可作为巨噬细胞的骨架蛋白[25] Obscurins encoded by this gene can be regarded as the cytoskeleton protein of macrophages
5	EPPK1	BLV-miR-B3-5p	920	在细胞骨架结构中起作用[26] It plays a role in the cytoskeletal structure
6	CDC42	BLV-miR-B4-5p	925	细胞骨架结构中发挥作用[20] It plays a role in the cytoskeletal structure
7	LTBP3	BLV-miR-B4-5p	1074	编码的蛋白质与转化生长因子β（TGF-β）形成复合物,在细胞外基质中发挥结构作用 The encoded protein forms a complex with transforming growth factor β (TGF-β) and plays a structural role in the extracellular matrix
8	DNAJC14	BLV-miR-B5-3p	316	是一种热休克蛋白,有助于Hsp70介导的蛋白质折叠[27] Is a heat shock protein that helps Hsp70-mediated protein folding
9	OBSCN	BLV-miR-B5-5p	1061	该基因编码的Obscurins蛋白可作为巨噬细胞的骨架蛋白[25] Obscurins encoded by this gene can be regarded as the cytoskeleton protein of macrophages
10	FNBP1L	BLV-miR-B2-5p	1469	该基因编码的蛋白质与CDC42和N-WASP结合,通过激活N-WASP-WIP复合物来促进CDC42诱导的肌动蛋白聚合 The protein encoded by this gene binds to CDC42 and N-WASP, and promotes CDC42-induced actin polymerization by activating the N-WASP-WIP complex
11	GAS2L1-208	BLV-miR-B2-5p	178	该蛋白结合细胞骨架的成分,并可能参与介导微管和微丝之间的相互作用;GAS2L1,一种微管和肌动蛋白结合蛋白,参与中心粒动力学和中心体分离[28];Gas2是一种生长停滞特异性蛋白,是微丝网络系统的组成部分[29] The protein binds to cytoskeletal components and is involved in mediating the interaction between microtubules and microfilaments. GAS2L1, a microtubule and actin binding protein, involved in centrosome dynamics and centrosome separation. Gas2 is a growth stagnation specific protein and a component of the microfilament network system
12	CLEC16A	BLV-miR-B2-3p	439	该基因编码包含C型凝集素结构域的家族的成员 The gene encodes a member of a family that contains the type C lectin domain
13	HERC5-201	BLV-miR-B2-5p	175	该基因是泛素连接酶HERC家族的成员,编码具有HECT结构域和5个RCC1重复序列的蛋白质 The gene is a member of the ubiquitin ligase HERC family that encodes proteins with HECT domains and five RCC1 repeats
14	MAST1	BLV-miR-B2-3p	304	该基因是微管相关丝氨酸/苏氨酸激酶（MAST）家族的成员。由该基因编码的蛋白质具有N端丝氨酸/苏氨酸激酶结构域 This gene is a member of the microtubule-associated serine/threonine kinase (MAST) family, and the protein encoded by this gene has an N-terminal serine/threonine kinase domain

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Table 5
表5
表5参与细胞增殖/凋亡的候选靶基因
Table 5Candidate target genes involved in cell proliferation

序号 Number	基因名称 Gene name	miRNA miRNA	评分 Score	与增殖关系 Relationship with proliferation
1	RPS6KA5	BLV-miR-B1-3p	618	抑制细胞增殖[30] Inhibiting of cell proliferation
2	SHPRH	BLV-miR-B1-3p	590	抑制细胞增殖[31] Inhibiting of cell proliferation
3	EVC	BLV-miR-B1-5p	2266	该基因发生突变可能通过下调Hh途径活性导致心肌细胞的增殖能力降低[32]。心肌细胞的抗细胞凋亡能力降低[32] Mutation of this gene may reduce the proliferation of cardiomyocytes by down-regulating Hh pathway activity. The anti-apoptotic ability of cardiomyocytes was decreased
4	BIRC6	BLV-miR-B2-3p	908	诱导细胞增殖[33]。抑制凋亡[34] Inducing cell proliferation and inhibiting apoptosis
5	EGFR	BLV-miR-B2-3p	1049	诱导细胞增殖 Inducing cell proliferation
6	COL16A1	BLV-miR-B3-5p	1031	通过上调内皮受体VEGFR1,VEGFR2和uPAR触发血管生成[35] Angiogenesis is triggered by upregulation of endothelial receptors VEGFR1, VEGFR2, and uPAR
7	KMT2D	BLV-miR-B3-5p	1203	下调抑制胃癌的增殖[36]。诱导其凋亡[36] Down-regulating gastric cancer proliferation and inducing apoptosis
8	TET3	BLV-miR-B4-3p	1691	细胞增殖[37] Cell proliferation
9	CDC42	BLV-miR-B4-5p	925	促进肿瘤生长[20] Promoting tumor growth
4	TRO	BLV-miR-B4-3p	1374	通过PKC-δ诱导人子宫内膜上皮细胞的凋亡 Apoptosis of human endometrial epithelial cells was induced by PKC-δ
5	COL1A2	BLV-miR-B4-3p	2543	通过PI3K-Akt信号通路抑制胃癌细胞凋亡[38] The apoptosis of gastric cancer cells was inhibited by pi3K-Akt signaling pathway
6	COL3A1	BLV-miR-B4-3p	2098	拮抗细胞凋亡功能[19] Antagonizing apoptosis
7	TIAL1-201	BLV-miR-B2-5p	180	该基因编码的蛋白质是RNA结合蛋白家族的成员,调节各种活动,包括翻译控制,剪接和凋亡 The gene encodes proteins that are members of the RNA-binding protein family and regulate various activities, including translation control, splicing, and apoptosis

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Table 6
表6
表6参与细胞分化的候选靶基因
Table 6Candidate target genes involved in cell differentiation

序号 Number	基因名称 Gene name	miRNA miRNA	评分 Score	与分化关系 Relationship with differentiation
1	MYO19	BLV-miR-B1-3p	596	确保细胞在分裂期间的彻底隔离[39] Ensure complete isolation of cells during division
2	RPS6KA5	BLV-miR-B1-3p	618	控制分化[40] Control of differentiation

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Table 7
表7
表7参与细胞迁移/侵袭的候选靶基因
Table 7Candidate target genes involved in cell migration / invasion

序号 Score	基因名称 Gene name	miRNA miRNA	评分 Score	与迁移关系 Relationship with migration
1	NOTCH3	BLV-miR-B1-3p	600	促进转移[41] Promoting metastasis
2	RICTOR	BLV-miR-B1-3p	592	促癌细胞转移[42] Promoting metastasis of cancer cells
3	TNXB	BLV-miR-B1-3p	747	具有抗黏附作用
4	RPS6KA5	BLV-miR-B1-3p	618	肿瘤侵袭有关[30] Relating to tumor invasion
5	FOCAD	BLV-miR-B3-3p	446	FOCAD编码一种在胶质瘤中具有肿瘤抑制功能的粘着斑蛋白[43] FOCAD encodes a focal adhesion proteins of an tumor inhibitory function in glioma
6	PLEKHG3	BLV-miR-B3-3p	444	激活细胞前端的肌动蛋白丝来增强极化细胞迁移[44] Activating the actin filaments at the cell front for enhancing the polarized migration of cells
7	EPPK1	BLV-miR-B3-5p	920	在伤口愈合期间加速角质形成细胞迁移 Promoting keratinocyte migration during wound healing
8	COL16A1	BLV-miR-B3-5p	1031	COL16A1在炎症晚期时在肠上皮下肌成纤维细胞（ISEMF）表面表达增加,导致细胞扩散[45] COL16A1 expression increasing in cell surface which led to cell diffusion
9	COL1A2	BLV-miR-B4-3p	2543	促进胃癌细胞迁移和侵袭[38] Promoting cell migration and invasion in gastric cancer cells
10	TRO	BLV-miR-B4-3p	1374	介导滋养细胞和子宫内膜上皮细胞之间的细胞黏附 It mediates cell adhesion between endometrial epithelial cells and trophoblast
11	CDC42	BLV-miR-B4-5p	925	影响细胞间黏附、肿瘤细胞形成过程的细胞迁移和侵袭[20] Affecting cell adhesion and cell migration and invasion during the tumor cell formation
12	EPN1	BLV-miR-B4-5p	1506	可能降低癌细胞稳定性 It may have decreased cancer cells stability
13	LTBP3	BLV-miR-B4-5p	1074	促进癌细胞传播过程中的早期转移[46] Promoting metastasis in cancer cells of early phase
14	DNAH17	BLV-miR-B5-3p	320	DNAH17编码动力蛋白轴索重链,参与细胞运动[47] DNAH17 encodes a dynein heavy chain motor protein and participates in cell motility
15	ATP11A	BLV-miR-B2-3p	563	编码的蛋白质是完整的膜,是结直肠癌异时转移的独立预测因子[48] The protein encoded was an independent predictor of colorectal cancer metastasis
16	ENPP7	BLV-miR-B2-3p	580	编码的蛋白质锚定在细胞膜中,它可能起到保护肠黏膜免受炎症和肿瘤发生的作用 It protects the intestinal mucosa against injuries inflammation and tumorigenesis to encoding protein which is anchored to the inner face of cellular membrane

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2.3 BLV miRNAs靶向调控人源白血病的候选靶基因

BLV属于反转录病毒科,丁型反转录病毒属,在分类学上与人类T淋巴细胞白血病病毒I型最为接近。而BLV miRNAs进入人体后仍然存在调控人源白血病细胞的可能性,BLV-miR-B4-3p作为患病牛体内表达量最高的BLV miRNA,在所有评分排名前十的候选靶基因中有两个靶基因可以在人急性淋巴细胞白血病（ALL）的发生发展中发挥作用,且均为BLV-miR-B4-3p的候选靶基因（表8）。

Table 8
表8
表8参与调控人急性淋巴细胞白血病的候选靶基因
Table 8Candidate target genes involved in the regulation of human acute lymphoblastic leukemia

序号 Score	基因名称 Gene name	miRNA miRNA	评分 Score	与白血病关系 Relationship with leukemia
1	COL1A1	BLV-miR-B4-3p	2603	ALL患者的骨骼发育异常与Col1A1 Sp1结合位点基因多态性存在相关性[49] There is a correlation between ALL patients have dysfunctional bone development and Col1A1 Sp1 binding site gene polymorphism
2	BCR	BLV-miR-B4-3p	320	ALL的融合转录本[50] The fusion transcripts of ALL

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2.4 BLV miRNAs靶向调控人源乳腺癌的候选靶基因

发现BLV miRNAs靶向调控的评分前十的候选靶基因中,有5个miRNA调控的7个候选靶基因在乳腺细胞的分化、迁移、侵袭生命活动中发挥了重要作用（表9）。

Table 9
表9
表9参与调控人乳腺癌的候选靶基因
Table 9Candidate target genes involved in the regulation of human breast cancer

序号 Score	基因名称 Gene name	miRNA miRNA	评分 Score	与乳腺癌关系 Relationship with breast cancer
1	NOTCH3	BLV-miR-B1-3p	600	在癌症转移过程中起作用[41] Playing a vital role in cancer metastasis
2	RICTOR	BLV-miR-B1-3p	592	在癌症转移过程中起作用[42, 51] Playing a vital role in cancer metastasis
3	RPS6KA5	BLV-miR-B1-3p	618	调节乳腺癌中腔细胞分化和转移性休眠,增加骨归巢和生长能力,与肿瘤侵袭有关[40] It regulates the differentiation of luminal cells and metastatic dormancy in breast cancer, increases bone homing and growth capacity and is related to tumor invasion
4	OBSCN	BLV-miR-B1-5p	2921	OBSCN缺失会导致细胞间接触被破坏,在体内外均会导致肿瘤的发生,迁移和侵袭[25] These cells contacts are disrupted by OBSCN defecting and leaded to cancer cell migration and invasion in vivo and in vitro
5	COL1A1	BLV-miR-B4-3p	2603	癌症来源的miR-218通过调控COL1A1在血液水平上乳腺癌向骨的转移过程起作用[49] MiR-218 of cancer work in transfer process with breast cancer metastasis to bone at the blood level by regulatory COL1A1
6	OBSCN	BLV-miR-B5-5p	1061	OBSCN缺失会导致细胞间接触被破坏,在体内外均会导致肿瘤的发生,迁移和侵袭[25] These cells contacts are disrupted by OBSCN defecting and leaded to cancer cell migration and invasion in vivo and in vitro
7	ATF3	BLV-miR-B2-5p	180	乳腺癌期间的病理状况下,已经观察到ATF3的持续和延长表达[52] ATF3 will prolonge and sustaine expression under pathological situations in breast cancer

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2.5 多个BLV miRNA共同靶向基因的最小自由能和二级结构

在评分前10的候选靶基因中,可以同时被多个BLV miRNA共同靶向的候选靶基因聚类于MUC5B、MUC12、MUC16基因（同属于黏蛋白家族）。我们通过RNAhybrid对这3个基因进行了二次预测验证的结果显示,这些BLV miRNA与靶基因结合的自由能均小于-15 kcal/mol（图2—4）。

图2

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图2靶向MUC5B的最小自由能和二级结构

红色核酸序列:候选靶基因序列;绿色核酸序列:miRNA序列;mfe:最小自由能
Fig. 2Minimum free energy and secondary structure of target MUC5B

Red nucleic acid sequence: Nucleic acid sequence of candidate target gene; Green nucleic acid sequence: Nucleic acid sequence of miRNA; mfe: minimum free energy

图3

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图3靶向MUC16的最小自由能和二级结构

红色核酸序列:候选靶基因序列;绿色核酸序列:miRNA序列;mfe:最小自由能
Fig. 3Minimum free energy and two stage structure of MUC16

Red nucleic acid sequence: Nucleic acid sequence of candidate target gene; Green nucleic acid sequence: Nucleic acid sequence of miRNA; mfe: Minimum free energy

图4

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图4靶向MUC12的最小自由能和二级结构

红色核酸序列:候选靶基因序列;绿色核酸序列:miRNA序列;mfe:最小自由能
Fig. 4Minimum free energy and secondary structure of target MUC12

Red nucleic acid sequence: Nucleic acid sequence of candidate target gene; Green nucleic acid sequence: Nucleic acid sequence of miRNA; mfe: Minimum free energy

3 讨论

20世纪90年代,URSIN等提出了一项前瞻性研究,研究了牛奶消费与癌症之间存在可能联系,评估了包括白血病、淋巴癌等癌症类型,虽然此项调查没有确定牛奶消费与癌症总发病率之间的关系,但是牛奶消费量（每天两杯）和淋巴器官癌症,特别是淋巴癌之间存在密切联系^[53],虽然它们之间是通过何种方式进行互作还需进一步研究,但这与我们所探究的BLV miRNAs通过食源途径跨界调控人源基因的风险可以相互印证。

笔者查询分析的70个候选靶基因中,36个候选靶基因的功能与肿瘤的发生发展存在相关性。这些候选靶基因所表达的蛋白在细胞的周期、信号转导、结构/骨架、增殖、凋亡、分化、迁移/侵袭功能中发挥重要作用。值得注意的是,调控细胞增殖与凋亡的候选靶基因均具有双向性,即促进与抑制细胞增殖和凋亡功能的基因共同存在,但对凋亡和增殖功能起抑制作用的基因相对较多,被miRNA靶向后可能增强细胞的代谢速度。有16个候选靶基因对细胞的迁移/侵袭功能产生影响,迁移/侵袭作为癌细胞最主要的功能表达形式,同样证实了BLV miRNAs与癌症存在相关性。

BLV-miR-B4-3p作为BLV编码的10种miRNA中,是患病牛体内表达量最高的一个^[10],与白血病相关miR 29a共享同一种子区域^[9],人急性淋巴细胞白血病的融合转录本同样作为BLV-miR-B4-3p的候选靶基因,提示BLV miRNA跨界进入人体后可能对人白血病的发生发展产生影响,虽然BLV与人类T淋巴细胞白血病病毒I型和Ⅱ（HTLV-1,2）同源性较高,但目前未见两种疾病间相关性的研究,这也可能是未来BLV的研究方向之一。但一直以来,BLV最受大家关注的公共卫生问题就是其与人乳腺癌之间的关系,虽然目前尚无证据表明BLV就是人源乳腺癌的重要致病因素之一,但近年来美国、澳大利亚、阿根廷等国家已经相继在乳腺癌患者的组织样品中检测到BLV基因片段^[7],BLV miRNAs作为BLV在机体内致病功能的主要发动者^{[9, 54]},我们的研究显示BLV miRNA在乳腺癌细胞的分化、迁移、侵袭过程中,均可以发挥重要作用,这一结论对BLV与人源乳腺癌之间相关性的研究有重要意义与实际价值。

BLV miRNAs的候选靶基因中,可以同时被多个BLV miRNA共同靶向的候选靶基因聚类于黏蛋白家族的MUC5B、MUC12、MUC16。其中MUC12可以编码一种完整的膜糖蛋白,可以在结肠表面的凝胶状分泌物中作为关键组成部分;虽然MUC5B是呼吸道黏蛋白的主要贡献者,MUC16是一种众所周知的卵巢癌标志物,但MUC5B在结肠中同样存在大量表达,MUC16也可以作为结肠癌的独立预后标志物^[55]。鉴于miRNA所存在的跨界调控机制^[16],而结肠作为机体吸收水分的重要肠段,我们推测BLV miRNAs极有可能通过靶向调控结肠组织细胞,破坏黏蛋白功能,进而跨界进入人体,并通过这一途径靶向调控人源基因。基于此,我们认为牛白血病病毒来源的miRNAs具有跨界调控人源基因的风险。

4 结论

外源性牛白血病病毒 miRNA可能跨界调控细胞周期、信号转导、结构/骨架、增殖、凋亡、分化、迁移/侵袭相关等细胞功能相关基因,破坏细胞结构。

牛白血病病毒 miRNA与人乳腺癌的相关性可能表现在人乳腺癌细胞的分化、迁移和侵袭过程中;而BLV-miR-B4-3p本身与白血病相关miR 29a共享同一种子区域,可能对人急性淋巴细胞白血病的发生发展造成影响。

外源性牛白血病病毒 miRNA具有靶向抑制黏蛋白基因（MUC5B、MUC12、MUC16）表达,通过破坏肠黏膜形成这一途径,跨界调控人源基因的风险。

参考文献 View Option 原文顺序
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[3] OTT S, JOHNSON R, WELLS S. Association between Bovine- Leukosis virus seroprevalence and herd-level productivity on US dairy farms
Preventive Veterinary Medicine, 2003,61(4):249-262.
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[4] ERSKINE R, BARLETT P C, BYREM T M, RENDER C L, FEBVAY C, HOUSEMAN J T. Association between bovine leukemia virus, production, and population age in Michigan dairy herds
Journal of Dairy Science, 2012,95:727-734.
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[5] FRIE M, SPORER K, WALLACE J, MAES R, SORDILLO L, BARTLETT P, COUSSENS P. Reduced humoral immunity and atypical cell-mediated immunity in response to vaccination in cows naturally infected with bovine leukemia virus
Veterinary Immunology and Immunopathology, 2016,182:125-135.
[本文引用: 1]

[6] MCCLURE H M, KEELING M E, CUSTER R P, MARSHAK R R, ABT D A, FERRER J F. Erythroleukemia in two infant chimpanzees fed milk from cows naturally infected with the bovine C-type virus
Cancer Research, 1974,34(10):2745-2757.
[本文引用: 1]

[7] MARTINEZ C L, PAMELA L, NIETO F M, DOLCINI G L, CERIANI C. Can bovine leukemia virus be related to human breast cancer? A review of the evidence
Journal of Mammary Gland Biology and Neoplasia, 2018,23(3):101-107.
[本文引用: 2]

[8] BUEHRING G C, DELANEY A, SHEN H, et al. Bovine leukemia virus discovered in human blood
BMC Infectious Diseases, 2019,19(1):297.
[本文引用: 1]

[9] KINCAID R P, BURKE J M, SULLIVAN C S, CHU D L C, RAZAVIAN N, SCHWARTZ D A, DEMKOVICH Z R, BATES M N. RNA virus microRNA that mimics a B-cell oncomiR
Proceedings of the National Academy of Sciences of the United States of America, 2012,109(8):3077-3082.
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[10] NICOLAS R, MéLANIE M, KEITH D, HARUKO T, FLORIAN C, YVETTE C, CéLINE V, FRANCK M, ERIC W, ARSèNE B, MICHEL G, ANNE V. Deep sequencing reveals abundant noncanonical retroviral microRNAs in B-cell leukemia/lymphoma
Proceedings of the National Academy of Sciences of the United States of America, 2013,110(6):2306-2311.
[本文引用: 3]

[11] MOULES V, POMIER C, SIBON D, GABET A S, REICHERT M, KERKHOFS P, WILLEMS L, MORTREUX F, WATTEL E. Fate of premalignant clones during the asymptomatic phase preceding lymphoid malignancy
Cancer Research, 2005,65(4):1234-1243.
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[12] MERIMI M, KLENER P, SZYNAL M, CLEUTER Y, KERKHOFS P, BURNY A, MARTIAT P, VAN DEN BROEKE A. Suppression of viral gene expression in bovine leukemia virus-associated B-cell malignancy: interplay of epigenetic modifications leading to chromatin with a repressive histone code
Journal of Virology, 2007,81(11):5929-5939.
[本文引用: 1]

[13] SAFARI R, HAMAIDIA M, DE BROGNIEZ A, GILLET N, WILLEMS L. Cis-drivers and trans-drivers of bovine leukemia virus oncogenesis
Current Opinion in Virology, 2017,26:15-19.
[本文引用: 1]

[14] GILLET N A, HAMAIDIA M, DE BROGNIEZ A, GUTIéRREZ G, RENOTTE N, REICHERT M, TRONO K, WILLEMS L. Bovine leukemia virus small noncoding rnas are functional elements that regulate replication and contribute to oncogenesis in vivo
PLoS Pathogens, 2016,12(4):e1005588.
[本文引用: 1]

[15] ROSEWICK N, DURKIN K, ARTESI M, MAR?AIS A, HAHAUT V, GRIEBEL P, ARSIC N, AVETTAND-FENOEL V, BURNY A, CHARLIER C, HERMINE O, GEORGES M, VAN DEN BROEKE A. Cis-perturbation of cancer drivers by the HTLV-1/BLV proviruses is an early determinant of leukemogenesis
Nature Communications, 2017,8(1):15264.
[本文引用: 1]

[16] ZHANG L, HOU D, LI D, ZHU L Y, ZHANG Y J, LI J, BIAN Z, LIANG X Y, CAI X, YIN Y, WANG C, ZHANG T F, ZHU D H, ZHANG D M, XU J, CHEN Q, BA Y, LIU J, ZHANG C Y. Exogenous plant MIR168a specifically targets mammalian LDLRAP1: evidence of cross-kingdom regulation by microRNA
Cell Research, 2011,22:107-126.
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[17] IZUMI H, TSUDA M, SATO Y, KOSAKA N, OCHIYA T, IWAMOTO H, NAMBA K, TAKEDA Y. Bovine milk exosomes contain microRNA and mRNA and are taken up by human macrophages
Journal of Dairy Science, 2015,98(5):2920-2933.
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[18] REHMSMEIER M, STEFFEN P, HOCHSMANN M, GIEGERICH R. Fast and effective prediction of microRNA/target duplexes
RNA -A Publication of The RNA Society, 2004,10(10):1507-1517.
[本文引用: 1]

[19] MUKHERJEE R, MAJUMDER P, CHAKRABARTI O. MGRN1- mediated ubiquitination of alpha-tubulin regulates microtubule dynamics and intracellular transport
Traffic, 2017,18(12):791-807.
DOI:10.1111/tra.12527URLPMID:28902452 [本文引用: 2]
MGRN1-mediated ubiquitination of alpha-tubulin regulates microtubule stability and mitotic spindle positioning in mitotic cells. This study elucidates the effect of MGRN1-mediated ubiquitination of alpha-tubulin in interphase cells. Here, we show that MGRN1-mediated ubiquitination regulates dynamics of EB1-labeled plus ends of microtubules. Intracellular transport of mitochondria and endosomes are affected in cultured cells where functional MGRN1 is depleted. Defects in microtubule-dependent organellar transport are evident in cells where noncanonical K6-mediated ubiquitination of alpha-tubulin by MGRN1 is compromised. Loss of MGRN1 has been previously correlated with late-onset spongiform neurodegeneration. Mislocalised cytosolically exposed PrP ((Ctm) PrP) interacts with MGRN1 leading to its loss of function. Expression of (Ctm) PrP generating mutants of PrP[PrP(A117V) and PrP(KHII)] lead to decrease in MGRN1-mediated ubiquitination of alpha-tubulin and intracellular transport defects. Brain lysates from PrP(A117V) transgenic mice also indicate loss of tubulin polymerization as compared to non-transgenic controls. Depletion of MGRN1 activity may hamper physiologically important processes like mitochondrial movement in neuronal processes and intracellular transport of ligands through the endosomal pathway thereby contributing to the pathogenesis of neurodegeneration in certain types of prion diseases.

[20] XIAO X H, LV L C, DUAN J, WU Y M, HE S J, HU Z Z, XIONG L X. Regulating Cdc42 and its signaling pathways in cancer: Small molecules and microrna as new treatment candidates
Molecules, 2018,23(4):787.
[本文引用: 4]

[21] FeLBERBAUM-CORTI M, MOREL E, CAVALLI V, VILBOIS F, GRUENBERG J. The redox sensor TXNL1 plays a regulatory role in fluid phase endocytosis
PLoS ONE, 2007,2(11):e1144.
DOI:10.1371/journal.pone.0001144URLPMID:17987124 [本文引用: 1]
BACKGROUND: Small GTPases of the Rab family can cycle between a GTP- and a GDP-bound state and also between membrane and cytosol. The latter cycle is mediated by the Guanine Nucleotide Dissociation Inhibitor GDI, which can selectively extract GDP-bound Rab proteins from donor membranes, and then reload them on target membranes. In previous studies, we found that capture of the small GTPase Rab5, a key regulator of endocytic membrane traffic, by GDI is stimulated by oxidative stress via p38MAPK, resulting in increased fluid phase endocytosis. METHODOLOGY/PRINCIPAL FINDINGS: When purifying the GDI stimulating activity we found that that it copurified with a high MW protein complex, which included p38MAPK. Here we report the identification and characterization of another component of this complex as the thioredoxin-like protein TXNL1. Our observations indicate that TXNL1 play a selective role in the regulation of fluid phase endocytosis, by controlling GDI capacity to capture Rab5. CONCLUSIONS/SIGNIFICANCE: Oxidants, which are known to cause cellular damage, can also trigger signaling pathways, in particular via members of the thioredoxin family. We propose that TXNL1 acts as an effector of oxidants or a redox sensor by converting redox changes into changes of GDI capacity to capture Rab5, which in turn modulates fluid phase endocytosis.

[22] LIU Z, ZHENG Y. A requirement for epsin in mitotic membrane and spindle organization
Journal of Cell Biology, 2009,186(4):473-480.
[本文引用: 1]

[23] DORNIER E, COUMAILLEAU F, OTTAVI J F, et al. TspanC8 tetraspanins regulate ADAM10 / Kuzbanian trafficking and promote Notch activation in flies and mammals
Journal of Cell Biology, 2012,199(3):481-496.
[本文引用: 1]

[24] QUINTERO O A, DIVITO M M, ADIKES R C, KORTAN M B, CASE L B, LIER A J, PANARETOS N S, SLATER S Q, RENGARAJAN M, FELIU M, CHENEY R E. Human Myo19 is a novel myosin that associates with mitochondria
Current Biology, 2009,19(23):2008-2013.
[本文引用: 1]

[25] SHRIVER M, STROKA K M, VITOLO M I, MARTIN S, HUSO D L, KONSTANTOPOULOS K, KONTROGIANNI-KONSTANTOPOULOS A. Loss of giant obscurins from breast epithelium promotes epithelial- to-mesenchymal transition, tumorigenicity and metastasis
Oncogene, 2015,34(32):4248-4259.
URLPMID:25381817 [本文引用: 4]

[26] JANG S I, KALININ A, TAKAHASHI K, MAREKOV L N, STEINERT P M. Characterization of human epiplakin: RNAi-mediated epiplakin depletion leads to the disruption of keratin and vimentin IF networks
Journal of Cell Science, 2005,118(Pt 4):781-793.
[本文引用: 1]

[27] JUNG J, KIM J, ROH S H, JUN I, SAMPSON R D, GEE H Y, CHOI J Y, LEE M G. The HSP70 co-chaperone DNAJC14 targets misfolded pendrin for unconventional protein secretion
Nature Communications, 2016,7:11386.
URLPMID:27109633 [本文引用: 1]

[28] AU F K, JIA Y, JIANG K, GRIGORIEV I, HAU B K, SHEN Y, DU S, AKHMANOVA A, QI R Z. GAS2L1 Is a centriole-associated protein required for centrosome dynamics and disjunction
Developmental Cell, 2017,40(1):81-94.
URLPMID:28017616 [本文引用: 1]

[29] BRANCOLINI C, BOTTEGA S, SCHNEIDER C. Gas2, a growth arrest-specific protein, is a component of the microfilament network system
Journal of Cell Biology, 1992,117(6):1251-1261.
[本文引用: 1]

[30] FU X, FAN X, HU J, ZOU H, CHEN Z, LIU Q, NI B, TAN X, SU Q, WANG J, WANG L, WANG J. Overexpression of MSK1 is associated with tumor aggressiveness and poor prognosis in colorectal cancer
Digestive and Liver Disease, 2017,49(6):683-691.
DOI:10.1016/j.dld.2017.02.009URLPMID:28314603 [本文引用: 2]
BACKGROUND/AIMS: Mitogen- and stress-activated protein kinase 1 (MSK1) has recently been implicated in cell proliferation and neoplastic transformation. However, the involvement of MSK1 in colorectal cancer (CRC) has not been addressed. This study aimed to evaluate the expression and potential functions of MSK1 in CRC. METHODS: The MSK1 expression was investigated by immunohistochemistry, western blot and reverse transcription-polymerase chain reaction. The associations between clinicopathological characteristics and MSK1 expression were assessed. Kaplan-Meier analysis and Cox regression models were carried out. CRC cells with MSK1 knockdown or overexpression were generated. A range of experiments were performed to demonstrate MSK1's role in CRC. RESULTS: MSK1 was overexpressed in 148 out of 329 CRC patients. CRC patients with high MSK1 expression had shorter overall survival than those with low MSK1 (P=0.033), especially among patients with stage III tumors (P=0.005). Knockdown of MSK1 in CRC cells suppressed cell proliferation, anchorage-independent growth, migration and invasion, and promoted 5-fluorouracil chemosensitivity and intracellular NADP(+)/NADPH ratio. However, overexpression of MSK1 had the opposite effects. CONCLUSIONS: Overexpression of MSK1 is associated with poor prognosis in CRC and is connected to tumor aggressiveness. MSK1 is a potential target for new therapies and a candidate of biomarker for prognosis.

[31] ZHANG M, HUANG N, YANG X, LUO J, YAN S, XIAO F, CHEN W, GAO X, ZHAO K, ZHOU H, LI Z, MING L, XIE B, ZHANG N. A novel protein encoded by the circular form of the SHPRH gene suppresses glioma tumorigenesis
Oncogene, 2018,37(13):1805-1814.
URLPMID:29343848 [本文引用: 1]

[32] LIU F, LIU X, XU Z, YUAN P, ZHOU Q, JIN J, YAN X, XU Z, CAO Q, YU J, CHENG Y, WAN R, HONG K. Molecular mechanisms of Ellisvan Creveld gene variations in ventricular septal defect
Molecular Medicine Reports, 2018,17(1):1527-1536.
DOI:10.3892/mmr.2017.8088URLPMID:29257216 [本文引用: 2]
The Ellis-van Creveld (EVC) gene is associated with various congenital heart diseases. However, studies on EVC gene variations in ventricular septal defect (VSD) and the underlying molecular mechanisms are sparse. The present study detected 11 singlenucleotide polymorphisms (SNPs) in 65 patients with VSD and 210 control patients from the Chinese Han population. Of the identified SNPs only the c.1727G>A SNP site was positively associated with the development of VSD (P<0.007). A known mutation, c.343C>G, was also identified, which causes a leucine to valine substitution at amino acid 115 of the EVC protein (p.L115V). The results of functional prediction indicated that c.343C>G may be a pathogenic mutation. In addition, in NIH3T3 mouse embryonic fibroblast cells, the EVC c.343C>G mutation significantly decreased cell proliferation and increased apoptosis. Further investigation demonstrated that in NIH3T3 cells, overexpression of EVC c.343C>G mutation reduced the binding between EVC and smoothened, which further downregulated the activity of the hedgehog (Hh) signaling pathway and the expression of downstream cyclin D1 and Bcell lymphoma 2 proteins with SAG. The c.1727G>A SNP of the EVC gene increased VSD susceptibility in patients from the Chinese Han population. The molecular mechanism underlying the development of VSD induced by the EVC c.343C>G mutation may be due to a reduction in the antiapoptotic and proliferative abilities of cardiomyocytes via downregulation of Hh pathway activity. The results of the present study may provide novel targets for the diagnosis and treatment of patients with VSD.

[33] WANG Z, LUO H, FANG Z, FAN Y, LIU X, ZHANG Y, RUI S, CHEN Y, HONG L, GAO J, ZHANG M. MiR-204 acts as a potential therapeutic target in acute myeloid leukemia by increasing BIRC6-mediated apoptosis
BMB Reports, 2018,51(9):444-449.
URLPMID:29764561 [本文引用: 1]

[34] MCCLURE H M, KEELING M E, CUSTER R P, MARSHAK R R, ABT D A, FERRER J F. Erythroleukemia in two infant chimpanzees fed milk from cows naturally infected with the bovine C-type virus
Cancer Research, 1974,34(10):2745-2757.
[本文引用: 1]

[35] BEDAL K B, GRASSEL S, SPANIER G, REICHERT T E, BAUER R J. The NC11 domain of human collagen XVI induces vasculogenic mimicry in oral squamous cell carcinoma cells
Carcinogenesis, 2015,36(11):1429-1439.
DOI:10.1093/carcin/bgv141URLPMID:26424749 [本文引用: 1]
Collagen XVI, a fibril-associated collagen with interrupted triple helix (FACIT) collagen, is involved in oral squamous cell carcinoma (OSCC) and glioblastoma progression. The NC11 domain of collagen XVI has been described previously with a strong implication in physiological processes. We detected the non-collagenous (NC) 11-domain in supernatants of OSCC cells after recombinant expression of full-length collagen XVI and in sera from OSCC patients and healthy individuals. Stable expression of NC11-green fluorescent protein (GFP) fusion protein in OSCC cells initiated proliferation control and block of anchorage-independent growth. Moreover, the NC11 domain triggered the generation of tubular-like net structures on laminin-rich matrix in contrast to mock-GFP control cells and cells expressing full-length collagen XVI. Taqman(R) quantitative PCR and diaminobenzidine staining in 2D- and 3D cell culture revealed a significantly increased gene and protein expression of VEGFR1, VEGFR2 and uPAR in recombinant NC11-GFP-expressing cells. Specific VEGF receptor inhibition with Axitinib or fetal calf serum heat inactivation prevented formation of tubular-like net structures. Accordantly, NC11-GFP coated culture slides led to an increase of focal adhesion contact formation and the upregulation of VEGFR1 and uPAR in three different non-transfected OSCC cell lines. In summary, we suggest that the NC11 domain of collagen XVI is a potential biomarker for OSCC and triggers vasculogenic mimicry via upregulation of endothelial receptors VEGFR1, VEGFR2 and uPAR in 2D- and 3D OSCC cell culture conditions.

[36] XIONG W, DENG Z, TANG Y, DENG Z, LI M. Downregulation of KMT2D suppresses proliferation and induces apoptosis of gastric cancer
Biochemical and Biophysical Research Communications, 2018,504(1):129-136.
[本文引用: 2]

[37] FB U B, CAU L, TAFAZZOLI A, MECHIN M C, WOLF S, ROMANO M T, VALENTIN F, WIEGMANN H, HUCHENQ A, KANDIL R, et al. Mutations in three genes encoding proteins involved in hair shaft formation cause uncombable hair syndrome
American Journal of Human Genetics, 2016,99(6):1292-1304.
URLPMID:27866708 [本文引用: 1]

[38] AO R, GUAN L, WANG Y, WANG J N. Silencing of COL1A2, COL6A3, and THBS2 inhibits gastric cancer cell proliferation, migration, and invasion while promoting apoptosis through the PI3k-Akt signaling pathway
Journal of Cellular Biochemistry, 2018,119(6):4420-4434.
URLPMID:29143985 [本文引用: 2]

[39] ROHN J L, PATEL J V, NEUMANN B, BULKESCHER J, MCHEDLISHVILI N, MCMULLAN R C, QUINTERO O A, ELLENBERG J, BAUM B. Myo19 ensures symmetric partitioning of mitochondria and coupling of mitochondrial segregation to cell division
Current Biology, 2014,24(21):2598-2605.
DOI:10.1016/j.cub.2014.09.045URLPMID:25447992 [本文引用: 1]
During animal cell division, an actin-based ring cleaves the cell into two. Problems with this process can cause chromosome missegregation and defects in cytoplasmic inheritance and the partitioning of organelles, which in turn are associated with human diseases. Although much is known about how chromosome segregation is coupled to cell division, the way organelles coordinate their inheritance during partitioning to daughter cells is less well understood. Here, using a high-content live-imaging small interfering RNA screen, we identify Myosin-XIX (Myo19) as a novel regulator of cell division. Previously, this actin-based motor was shown to control the interphase movement of mitochondria. Our analysis shows that Myo19 is indeed localized to mitochondria and that its silencing leads to defects in the distribution of mitochondria within cells and in mitochondrial partitioning at division. Furthermore, many Myo19 RNAi cells undergo stochastic division failure--a phenotype that can be mimicked using a treatment that blocks mitochondrial fission and rescued by decreasing mitochondrial fusion, implying that mitochondria can physically interfere with cytokinesis. Strikingly, using live imaging we also observe the inappropriate movement of mitochondria to the poles of spindles in cells depleted for Myo19 as they enter anaphase. Since this phenocopies the results of an acute loss of actin filaments in anaphase, these data support a model whereby the Myo19 actin-based motor helps to control mitochondrial movement to ensure their faithful segregation during division. The presence of DNA within mitochondria makes their inheritance an especially important aspect of symmetrical cell division.

[40] GAWRZAK S, RINALDI L, GREGORIO S, ARENAS E J, SALVADOR F, UROSEVIC J, FIGUERAS-PUIG C, ROJO F, DEL BARCO BARRANTES I, CEJALVO J M, et al. MSK1 regulates luminal cell differentiation and metastatic dormancy in ER(+) breast cancer
Nature Cell Biology, 2018,20(2):211-221.
[本文引用: 2]

[41] LEONTOVICH A A, JALALIRAD M, SALISBURY J L, MILLS L, HADDOX C, SCHROEDER M, TUMA A, GUICCIARDI M E, ZAMMATARO L, GAMBINO M W, et al. NOTCH3 expression is linked to breast cancer seeding and distant metastasis
Breast Cancer Reserach, 2018,20(1):105.
[本文引用: 2]

[42] EL SHAMIEH S, SALEH F, MOUSSA S, KATTAN J, FARHAT F. RICTOR gene amplification is correlated with metastasis and therapeutic resistance in triple-negative breast cancer
Pharmacogenomics, 2018,19(9):757-760.
DOI:10.2217/pgs-2018-0019URLPMID:29790419 [本文引用: 2]
Triple-negative breast cancer (TNBC) is characterized by its aggressive behavior, metastasis and lack of targeted therapies. Herein, we discuss the clinical, histopathological and genetic profile of a woman diagnosed with TNBC. Since the patient had no durable response to chemotherapy, a genetic profiling was carried out. Next-generation sequencing analysis of 592 genes showed a missense mutation, p.E545A in PIK3CA, thus the patient was started on the mTOR inhibitor everolimus, in combination with exemestane, which controlled her pain; however, the disease progressed aggressively. More importantly, next-generation sequencing analysis showed a RICTOR gene amplification (eight copies) suggesting that RICTOR promotes the genesis of TNBC. We conclude that determining regulators of RICTOR and furthermore, their inhibitors might decrease cancer cells proliferation rate in patients with TNBC.

[43] BROCKSCHMIDT A, TROST D, PETERZIEL H, ZIMMERMANN K, EHRLER M, GRASSMANN H, PFENNING P N, WAHA A, WOHLLEBER D, BROCKSCHMIDT F F, et al. KIAA1797/FOCAD encodes a novel focal adhesion protein with tumour suppressor function in gliomas
Brain, 2012,135(Pt 4):1027-1041.
DOI:10.1093/brain/aws045URLPMID:22427331 [本文引用: 1]
In a strategy to identify novel genes involved in glioma pathogenesis by molecular characterization of chromosomal translocation breakpoints, we identified the KIAA1797 gene, encoding a protein with an as yet undefined function, to be disrupted by a 7;9 translocation in a primary glioblastoma culture. Array-based comparative genomic hybridization detected deletions involving KIAA1797 in around half of glioblastoma cell lines and glioblastomas investigated. Quantification of messenger RNA levels in human tissues demonstrated highest KIAA1797 expression in brain, reduced levels in all glioblastoma cell lines and most glioblastomas and similar levels in glial and neuronal cells by analysis of different hippocampal regions from murine brain. Antibodies against KIAA1797 were generated and showed similar protein levels in cortex and subcortical white matter of human brain, while levels were significantly reduced in glioblastomas with KIAA1797 deletion. By immunofluorescence of astrocytoma cells, KIAA1797 co-localized with vinculin in focal adhesions. Physical interaction between KIAA1797 and vinculin was demonstrated via co-immunoprecipitation. Functional in vitro assays demonstrated a significant decrease in colony formation, migration and invasion capacity of LN18 and U87MG glioma cells carrying a homozygous KIAA1797 deletion ectopically expressing KIAA1797 compared with mock-transduced cells. In an in vivo orthotopic xenograft mouse model, U87MG tumour lesions expressing KIAA1797 had a significantly reduced volume compared to tumours not expressing KIAA1797. In summary, the frequently deleted KIAA1797 gene encodes a novel focal adhesion complex protein with tumour suppressor function in gliomas, which we name 'focadhesin'. Since KIAA1797 genetic variation has been implicated in Alzheimer's disease, our data are also relevant for neurodegeneration.

[44] NGUYEN T T, PARK W S, PARK B O, KIM C Y, OH Y, KIM J M, CHOI H, KYUNG T, KIM C H, LEE G, et al. PLEKHG3 enhances polarized cell migration by activating actin filaments at the cell front
Proceedings of the National Academy of Sciences of the United States of America, 2016,113(36):10091-10096.
[本文引用: 1]

[45] RATZINGER S, EBLE J A, PASOLDT A, OPOLKA A, ROGLER G, GRIFKA J, GRASSEL S. Collagen XVI induces formation of focal contacts on intestinal myofibroblasts isolated from the normal and inflamed intestinal tract
Matrix Biology, 2010,29(3):177-193.
DOI:10.1016/j.matbio.2009.11.004URLPMID:19931388 [本文引用: 1]
In Crohn's disease (CD) the stress-shield of intestinal subepithelial myofibroblasts (ISEMF) provided by intact tissue is disturbed due to inflammation and thus, cells start with remodelling activities. This is characterized by increased numbers of collagen-producing ISEMF causing an uncontrolled, irreversible wound-healing response to the chronic inflammation of the gastrointestinal tract. Reconstitution of the original ECM leads ISEMF to exit this cycle. In contrast, during fibrosis, ISEMF persist. It is known that ISEMF produce and deposit collagen types I, III, IV and V; however synthesis and the role of fibrillar peripheral molecules like collagen type XVI have not been addressed yet. Here, we have analyzed the distribution of collagen XVI in the normal and inflamed bowel wall, its gene and protein expression by ISEMF of different inflammation stages, the cell-matrix interactions in different phases of the inflammatory process and their effect on cell spreading, proliferation and migration. Collagen XVI is deposited in the submucosa of the intestinal wall where it co-localizes with fibrillin-1 and integrin alpha1. ISEMF reveal increasing gene and protein expression of collagen XVI concurrent to increasing inflammation. ISEMF reveal more mature focal adhesion contacts when seeded on collagen XVI resulting in an extensive cell spreading. This involves recruitment of alpha1beta1 integrin, which shows increased cell surface expression on ISEMF in late stages of inflammation. We assume that collagen XVI promotes persistence of ISEMF in the normal and, even stronger in the inflamed bowel wall by stabilizing focal adhesion contacts via cell-matrix interaction preferentially through recruitment of alpha1ss1 integrin into the tips of the focal adhesion contacts.

[46] DERYUGINA E I, ZAJAC E, ZILBERBERG L, MURAMATSU T, JOSHI G, DABOVIC B, RIFKIN D, QUIGLEY J P. LTBP3 promotes early metastatic events during cancer cell dissemination
Oncogene, 2018,37(14):1815-1829.
DOI:10.1038/s41388-017-0075-1URLPMID:29348457 [本文引用: 1]
Latent transforming growth factor beta (TGFbeta)-binding proteins (LTBPs) are important for the secretion, activation, and function of mature TGFbeta, especially so in cancer cell physiology. However, specific roles of the LTBPs remain understudied in the context of the primary tumor microenvironment. Herein, we investigated the role of LTBP3 in the distinct processes involved in cancer metastasis. By using three human tumor cell lines of different tissue origin (epidermoid HEp-3 and prostate PC-3 carcinomas and HT-1080 fibrosarcoma) and several metastasis models conducted in both mammalian and avian settings, we show that LTBP3 is involved in the early dissemination of primary cancer cells, namely in the intravasation step of the metastatic cascade. Knockdown of LTBP3 in all tested cell lines led to significant inhibition of tumor cell intravasation, but did not affect primary tumor growth. LTBP3 was dispensable in the late steps of carcinoma cell metastasis that follow tumor cell intravasation, including vascular arrest, extravasation, and tissue colonization. However, LTBP3 depletion diminished the angiogenesis-inducing potential of HEp-3 cells in vivo, which was restorable by exogenous delivery of LTBP3 protein. A similar compensatory approach rescued the dampened intravasation of LTBP3-deficient HEp-3 cells, suggesting that LTBP3 regulates the induction of the intravasation-supporting angiogenic vasculature within developing primary tumors. Using our recently developed microtumor model, we confirmed that LTBP3 loss resulted in the development of intratumoral vessels with an abnormal microarchitecture incompatible with efficient intravasation of HEp-3 carcinoma cells. Collectively, these findings demonstrate that LTBP3 represents a novel oncotarget that has distinctive functions in the regulation of angiogenesis-dependent tumor cell intravasation, a critical process during early cancer dissemination. Our experimental data are also consistent with the survival prognostic value of LTBP3 expression in early-stage head and neck squamous cell carcinomas, further indicating a specific role for LTBP3 in cancer progression toward metastatic disease.

[47] ZHU C, YANG Q, XU J, ZHAO W, ZHANG Z, XU D, ZHANG Y, ZHAO E, ZHAO G. Somatic mutation of DNAH genes implicated higher chemotherapy response rate in gastric adenocarcinoma patients
Journal of Translational Medicine, 2019,17(1):109.
DOI:10.1186/s12967-019-1867-6URLPMID:30944005 [本文引用: 1]
BACKGROUND: The dynein axonemal heavy chain (DNAH) family of genes encode the dynein axonemal heavy chain, which is involved in cell motility. Genomic variations of DNAH family members have been frequently reported in diverse kinds of malignant tumors. In this study, we analyzed the genomic database to evaluate the mutation status of DNAH genes in gastric adenocarcinoma and further identified the significance of mutant DNAH genes as effective molecular biomarkers for predicting chemotherapy response in gastric cancer patients. METHODS: We analyzed the clinical and genomic data of gastric cancer patients published in The Cancer Genome Atlas (TCGA) project. Data on chemotherapy response, overall survival (OS) and chemotherapy-free survival were retrieved. Then, we verified the results via targeted sequencing of gastric cancer patients with similar clinical characteristics but different chemotherapeutic outcomes. RESULTS: In total, 132 gastric adenocarcinoma patients undergoing chemotherapy treatment from TCGA were included in our study. Somatic mutations in all 13 members of the DNAH family of genes were associated with different chemotherapy responses. Compared with patients with wild-type DNAH genes (n = 59), a significantly higher proportion of those with mutations in DNAH genes (n = 73) (55.9% vs 80.8%) responded to chemotherapy (P = 0.002). Moreover, DNAH mutations were correlated with significantly better OS (P = 0.027), chemotherapy-free survival (P = 0.027), fluoropyrimidine-free survival (P = 0.048) and platinum-free survival (P = 0.014). DNAH mutation status was an independent risk factor for OS (P = 0.015), chemotherapy-free survival (P = 0.015) and platinum-free survival (P = 0.011). We identified somatic mutations in 27 (42.2%) of the 64 stage III gastric adenocarcinoma patients receiving fluoropyrimidine-based chemotherapy by targeted exon sequencing with strict screening conditions. In our own cohort, a significantly higher proportion of patients (n = 32) with DNAH mutations than patients with wild-type DNAH genes (n = 32) had a good prognosis (OS > 48 months) (70.4% vs 35.1%) (P = 0.005). CONCLUSIONS: Dynein axonemal heavy chain gene mutations contribute positively to chemotherapy sensitivity in gastric cancer patients.

[48] MIYOSHI N, ISHII H, MIMORI K, TANAKA F, NAGAI K, UEMURA M, SEKIMOTO M, DOKI Y, MORI M. ATP11A is a novel predictive marker for metachronous metastasis of colorectal cancer
Oncology Reports, 2010,23(2):505-510.
URLPMID:20043114 [本文引用: 1]

[49] ZHANG Z, FANG C, WANG Y, ZHANG J, YU J, ZHANG Y, WANG X, ZHONG J. COL1A1: A potential therapeutic target for colorectal cancer expressing wild-type or mutant KRAS
International Journal of Oncology, 2018,53(5):1869-1880.
DOI:10.3892/ijo.2018.4536URLPMID:30132520 [本文引用: 2]
Colorectal cancer (CRC) treatment primarily relies on chemotherapy along with surgery, radiotherapy and, more recently, targeted therapy at the late stages. However, chemotherapeutic drugs have high cytotoxicity, and the similarity between the effects of these drugs on cancerous and healthy cells limits their wider use in clinical settings. Targeted monoclonal antibody treatment may compensate for this deficiency. Epidermal growth factor receptor (EGFR)targeted drugs have a positive effect on CRC with intact KRAS proto-oncogene GTPase (KRAS or KRASWT), but may be ineffective or harmful in patients with KRAS mutations (KRASMUT). Therefore, it is important to identify drug target genes that are uniformly effective with regards to KRASWT and KRASMUT CRC. The present study performed gene expression analysis, and identified 294 genes upregulated in KRASWT and KRASMUT CRC samples. Collagen type I alpha 1 (COL1A1) was identified as the hub gene through STRING and Cytoscape analyses. Consistent with results obtained from Oncomine, a cancer microarray database and web-based data-mining platform, it was demonstrated that the expression of COL1A1 was significantly upregulated in CRC tissues and cell lines regardless of KRAS status. Inhibition of COL1A1 in KRASWT and KRASMUT CRC cell lines significantly decreased cell proliferation and invasion. In addition, increased COL1A1 expression in CRC was significantly associated with serosal invasion, lymph metastases and hematogenous metastases. Taken together, the findings of the present study indicated that COL1A1 may serve as a candidate diagnostic biomarker and a promising therapeutic target for CRC.

[50] CHOPRA A, SONI S, VERMA D, KUMAR D, DWIVEDI R, VISHWANATHAN A, VISHWAKAMA G, BAKHSHI S, SETH R, GOGIA A, KUMAR L, KUMAR R. Prevalence of common fusion transcripts in acute lymphoblastic leukemia: A report of 304 cases
Asia-Pacific Journal of Clinical Oncology, 2015,11(4):293-298.
DOI:10.1111/ajco.12400URLPMID:26264145 [本文引用: 1]
AIM: Information about fusion transcripts in acute lymphoblastic leukemia (ALL) is used to risk-stratify patients, decide on the treatment and to detect minimal residual disease. This study was conducted to determine the frequency of common fusion transcripts BCR-ABL, TEL-AML1, MLL-AF4 and E2A-PBX1 for B-ALL and SIL-TAL1 for T-ALL as seen at a tertiary care center in India. METHODS: Up to 304 new cases of ALL (271 B-ALL and 33 T-ALL) diagnosed on morphology, cytochemistry and immunophenotyping were studied. All were screened for the common fusion transcripts by RT-PCR. RESULTS: Both our B- (218/271; 80.4%) and T-ALL (26/33; 78.8%) patients were largely children. In the B-ALL children, BCR-ABL was detected in 26/218 (11.9%), E2A-PBX1 in 13/218 (5.9%), TEL-AML1 in 16/218 (7.3%) and MLL-AF4 in 3/218 (1.4%) patients. Adult B-ALL cases had BCR-ABL in 15/53 (28.3%) and E2A-PBX in 2/53 (3.8%); however, no other fusion transcript was detected. SIL-TAL1 was found in four of 26 pediatric (15%) and zero of 7 adult T-ALL cases. CONCLUSION: The higher incidence of BCR-ABL and lower incidence of TEL-AML1 in our ALL patients, both in children and adults as compared with the West, suggests that patients in India may be biologically different. This difference may explain at least in part the higher relapse rate and poorer outcome in our B-ALL cases.

[51] SCHMIDT K M, DIETRICH P, HACKL C, GUENZLE J, BRONSERT P, WAGNER C, FICHTNER-FEIGL S, SCHLITT H J, GEISSLER E K, HELLERBRAND C, LANG S A. Inhibition of mTORC2/RICTOR impairs melanoma hepatic metastasis
Neoplasia, 2018,20(12):1198-1208.
DOI:10.1016/j.neo.2018.10.001URLPMID:30404068 [本文引用: 1]
Mammalian target of rapamycin complex 2 (mTORC2) with its pivotal component rapamycin-insensitive companion of mTOR (RICTOR) is the major regulator of AKT phosphorylation and is increasingly implicated in tumor growth and progression. In cutaneous melanoma, an extremely aggressive and highly metastatic disease, RICTOR overexpression is involved in tumor development and invasiveness. Therefore, we investigated the impact of RICTOR inhibition in melanoma cells in vitro and in vivo with special emphasis on hepatic metastasis. Moreover, our study focused on the interaction of tumor cells and hepatic stellate cells (HSC) which play a crucial role in the hepatic microenvironment. In silico analysis revealed increased RICTOR expression in melanoma cells and tissues and indicated higher expression in advanced melanoma stages and metastases. In vitro, transient RICTOR knock-down via siRNA caused a significant reduction of tumor cell motility. Using a syngeneic murine splenic injection model, a significant decrease in liver metastasis burden was detected in vivo. Moreover, stimulation of melanoma cells with conditioned medium (CM) from activated HSC or hepatocyte growth factor (HGF) led to a significant induction of AKT phosphorylation and tumor cell motility. Blocking of RICTOR expression in cancer cells diminished constitutive and HGF-induced AKT phosphorylation as well as cell motility. Interestingly, RICTOR blockade also led to an abrogation of CM-induced effects on AKT phosphorylation and motility in melanoma cells. In conclusion, these results provide first evidence for a critical role of mTORC2/RICTOR in melanoma liver metastasis via cancer cell/HSC interactions.

[52] ROHINI M, HARITHA MENON A, SELVAMURUGAN N. Role of activating transcription factor 3 and its interacting proteins under physiological and pathological conditions
International Journal of Biological Macromolecules, 2018,120(Pt A):310-317.
DOI:10.1016/j.ijbiomac.2018.08.107URLPMID:30144543 [本文引用: 1]
Activating transcription factor 3 (ATF3) is a stress-responsive factor that belongs to the activator protein 1 (AP-1) family of transcription factors. ATF3 expression is stimulated by various factors such as hypoxia, cytokines, and chemotherapeutic and DNA damaging agents. Upon stimulation, ATF3 can form homodimers or heterodimers with other members of the AP-1 family to repress or activate transcription. Under physiological conditions, ATF3 expression is transient and plays a pivotal role in controlling the expression of cell-cycle regulators and tumor suppressor, DNA repair, and apoptosis genes. However, under pathological conditions such as those during breast cancer, a sustained and prolonged expression of ATF3 has been observed. In this review, the structure and function of ATF3, its posttranslational modifications (PTM), and its interacting proteins are discussed with a special emphasis on breast cancer metastasis.

[53] URSIN G, BJELKE E, HEUCH I, VOLLSET S E. Milk consumption and cancer incidence: a Norwegian prospective study
British Journal of Cancer, 1990,61(3):454-459.
DOI:10.1038/bjc.1990.100URLPMID:2328215 [本文引用: 1]
Relationships between milk intake and cancer incidence were investigated after 11 1/2 years of follow-up of 15,914 individuals. A diagnosis of cancer was made in a total of 1,422 individuals. No association was established with total cancer incidence, in analyses adjusted for sex, age and residential characteristics. However, a strong positive association with milk consumption was observed for cancers of the lymphatic organs (odds ratio 3.4 for greater than or equal to 2 glasses per day vs less than 1; 95% confidence interval 1.4-8.2). An inverse association was found for cancer of the bladder. Kidney cancer and cancers of the female reproductive organs (except the uterine cervix) showed weak positive associations with milk intake.

[54] SANTANAM U, ZANESI N, EFANOV A, COSTINEAN S, PALAMARCHUK A, HAGAN J P, VOLINIA S, ALDER H, RASSENTI L, KIPPS T, CROCE C M, PEKARSKY Y. Chronic lymphocytic leukemia modeled in mouse by targeted miR-29 expression
Proceedings of the National Academy of Sciences of the United States of America, 2010,107(27):12210-12215.
[本文引用: 1]

[55] BJORKMAN K, MUSTONEN H, KAPRIO T, HAGLUND C, BOCKELMAN C. Mucin 16 and kallikrein 13 as potential prognostic factors in colon cancer: Results of an oncological 92-multiplex immunoassay
Tumour Biology, 2019,41(7):1010428319860728.
[本文引用: 1]