沉默FBI-1基因对三阴性乳腺癌细胞MDA-MB-231增殖和凋亡的影响
陈茂剑¹, 王丽², 杨伟萍¹, 覃庆洪¹, 谭启杏¹, 练斌¹, 韦长元^1,*
1广西医科大学附属肿瘤医院乳腺外科，南宁 530021；²柳州市人民医院乳腺外科，柳州 545006
摘要
本研究旨在观察短发夹状RNA (short hairpin RNA, shRNA)介导的人类免疫缺陷病毒短转录诱导物连接因子1 (FBI-1)基因沉默对三阴性乳腺癌细胞MDA-MB-231增殖和凋亡的影响。用qRT-PCR和Western blot分别检测FBI-1、Bcl-2、Bax、cleaved-Caspase 3和Survivin的mRNA和/或蛋白的表达水平；采用shRNA干扰技术沉默MDA-MB-231细胞中FBI-1基因的表达；用CCK-8法以及克隆形成实验检测细胞增殖；用流式细胞术检测细胞凋亡；用裸鼠皮下成瘤实验检测细胞的成瘤能力。结果显示，MDA-MB-231细胞的FBI-1 mRNA和蛋白的表达水平明显高于正常乳腺上皮细胞MCF-10A；经shRNA靶向沉默FBI-1基因表达后，细胞增殖能力明显降低，细胞凋亡率显著增高，伴随Bcl-2和Survivin蛋白表达明显下调、Bax蛋白表达显著上调和Caspase 3活化，MDA-MB-231细胞的裸鼠皮下成瘤能力受抑制。以上结果提示，靶向沉默FBI-1基因表达可以抑制MDA-MB-231细胞增殖，诱导细胞凋亡并抑制细胞的裸鼠皮下成瘤能力。


关键词： FBI-1; 短发夹状RNA; 三阴性乳腺癌; 细胞增殖; 细胞凋亡
分类号：R737.9

Effects of FBI-1 silencing on proliferation and apoptosis of triple-negative breast cancer cell line MDA-MB-231
CHEN Mao-Jian¹, Wang Li², YANG Wei-Ping¹, QIN Qing-Hong¹, TAN Qi-Xing¹, LIAN Bin¹, WEI Chang-Yuan^1,*
1Department of Breast Surgery, The Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China；²Department of Breast Surgery, Liuzhou People’s Hospital, Liuzhou 545006, China
Abstract
This work aimed to observe the effects of short hairpin RNA (shRNA)-silenced FBI-1 (factor that binds to the inducer of short transcripts of human immunodeficiency virus-1) on proliferation and apoptosis of triple-negative breast cancer cell line MDA-MB-231. qRT-PCR and Western blot analysis were applied to detect the mRNA and/or protein expression of FBI-1, Bcl-2, Bax, cleaved-Caspase 3 and Survivin. RNA interference method was used to silence FBI-1 expression in MDA-MB-231 cells. CCK-8 and colony formation assay were employed to detect the cell proliferation. Flow cytometry was employed for examining cell apoptosis. In vivo tumorigenicity of MDA-MB-231 cells was detected by tumor transplantation in nude mice. The results showed that the mRNA and protein expressions of FBI-1 were higher in MDA-MB-231 cells compared with those in normal human mammary epithelial cells MCF-10A. FBI-1 gene silencing inhibited proliferation and induced apoptosis of MDA-MB-231 cells in vitro, together with decreased Bcl-2 and Survivin protein expression, increased Bax protein expression and activated Caspase 3. Moreover, FBI-1 gene silencing
inhibited the tumorigenesis of MDA-MB-231 cells in vivo. These results suggest that silencing of FBI-1 gene inhibits proliferation, induces apoptosis and suppresses the tumorigenesis of MDA-MB-231 cells.


Key words: FBI-1; short hairpin RNA; triple-negative breast cancer; cell proliferation; cell apoptosis

收稿日期：2018-02-01　　录用日期：2018-08-14
通讯作者：韦长元　　E-mail: weicy63@aliyun.com
DOI: 10.13294/j.aps.2018.0056
引用本文：
陈茂剑, 王丽, 杨伟萍, 覃庆洪, 谭启杏, 练斌, 韦长元. 沉默FBI-1基因对三阴性乳腺癌细胞MDA-MB-231增殖和凋亡的影响[J]. 生理学报 2018; 70 (5): 497-503.
CHEN Mao-Jian, Wang Li, YANG Wei-Ping, QIN Qing-Hong, TAN Qi-Xing, LIAN Bin, WEI Chang-Yuan. Effects of FBI-1 silencing on proliferation and apoptosis of triple-negative breast cancer cell line MDA-MB-231. Acta Physiol Sin 2018; 70 (5): 497-503 (in Chinese with English abstract).



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