摘要： 目的 探究长链非编码RNA （lncRNA） ATP6V1B1-AS1对非小细胞肺癌增殖与侵袭的作用及其机制。方法 实时定量PCR （qRT-PCR）检测非小细胞肺癌细胞系（A549、SK-MES-1、1299、H460）及正常人支气管上皮细胞系（16-HBE）中ATP6V1B1-AS1的表达；从UCSC xena数据库下载癌症基因组图谱（TCGA）泛癌数据集，分析泛癌中ATP6V1B1-AS1表达情况。采用CCK8实验、Transwell实验检测对照组、过表达ATP6V1B1-AS1组、敲减ATP6V1B1-AS1组细胞增殖及侵袭能力；Western blotting检测侵袭相关蛋白E-cadherin、MMP9、Snail表达。通过生物信息学方法构建ATP6V1B1-AS1参加调控的内源竞争RNA （ceRNA）网络，分析ATP6V1B1-AS1对非小细胞肺癌增殖与侵袭可能的作用机制。利用基因本体（GO）数据库及京都基因和基因组数据库（KEGG）富集分析ATP6V1B1-AS1参与调控的生物学功能。结果 与正常人支气管上皮系比较，非小细胞肺癌细胞系A549、H460、H1299、SK-MES-1中ATP6V1B1-AS1的表达均显著增高（均P<0.05）。TCGA泛癌分析结果显示，ATP6V1B1-AS1在多种癌症中高表达，并且肺鳞癌及肺腺癌中高表达，差异有统计学意义（P<0.05）。A549细胞过表达ATP6V1B1-AS1后增殖和侵袭能力增强，SK-MES-1细胞敲减ATP6V1B1-AS1后增殖和侵袭能力减弱。与对照组比较，过表达ATP6V1B1-AS1后A549细胞E-cadherin蛋白表达减少，Snail和MMP9蛋白表达增加（P<0.05）；敲减ATP6V1B1-AS1后SK-MES-1细胞的E-cadherin蛋白表达增加，Snail和MMP9蛋白表达减少（P<0.05）。ceRNA网络调控图结果显示，miR-520a-5p、miR-526b-3p、miR-4524a-3p、miR-6730-5p可能是ATP6V1B1-AS1的下游基因，调控WNT3、PAICS等基因发挥ATP6V1B1-AS1促进非小细胞肺癌增殖、侵袭的作用。GO数据库功能富集分析结果显示，ATP6V1B1-AS1下游mRNA与腺体发展等功能相关，KEGG功能富集分析显示ATP6V1B1-AS1与嘌呤代谢等功能相关。结论 ATP6V1B1-AS1在非小细胞肺癌细胞系中高表达，并能够促进非小细胞肺癌的增殖、侵袭。ATP6V1B1-AS1可能通过ceRNA网络调控相应生物学行为。

lncRNA ATP6V1B1-AS1对非小细胞肺癌增殖与侵袭的作用及其机制

李依霖, 叶军, 何连栋, 王嘉俊, 许顺

中国医科大学附属第一医院胸外科, 沈阳 110001

收稿日期:2022-09-28出版日期:2023-01-30发布日期:2023-02-01
通讯作者:许顺E-mail:xushun610539@sina.com
作者简介:李依霖（1996-），男，硕士研究生.
基金资助:辽宁省科学技术基金博士启动基金（2021-BS-094）；沈阳市科学技术计划（20-205-4-043）


关键词: 非小细胞肺癌, 长链非编码RNA, ATP6V1B1-AS1, 增殖, 侵袭
Abstract: Objective To investigate the effects and mechanism of long non-coding RNA ATP6V1B1-AS1 on the proliferation and invasion of non-small cell lung cancer. Methods The expression of ATP6V1B1-AS1 in non-small cell lung cancer cell lines (A549, SK-MES-1, 1299, and H460) and normal bronchial epithelial cell lines (16-HBE) was detected by qRT-PCR. The Cancer Genome Atlas (TCGA) pan-carcinoma dataset was downloaded from the UCSC xena database to analyze the expression of ATP6V1B1-AS1 in pan-carcinoma. The CCK8 assay and Transwell assay were used to detect cell proliferation and invasion ability in the control group, the ATP6V1B1-AS1 overexpression group, and the ATP6V1B1-AS1 knockdown group. Western blotting detected the expression of invasion-related proteins E-cadherin, MMP9, and Snail. Bioinformatics methods were used to construct the competing endogenous RNA (ceRNA) network regulated by ATP6V1B1-AS1, and the possible mechanism of action of ATP6V1B1-AS1 on proliferation and invasion of non-small cell lung cancer was analyzed by ceRNA. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) database were used to analyze the biological function of ATP6V1B1-AS1 that may be involved in regulation. Results Compared with normal bronchial epithelial lines, the expression of ATP6V1B1-AS1 in non-small cell lung cancer cell lines (A549, H460, H1299, and SK-MES-1) was significantly increased (all P<0.05). The invasion ability of A549 cells was enhanced after overexpression of ATP6V1B1-AS1 and weakened after ATP6V1B1-AS1 knockdown of SK-MES-1 cells. Compared with the control group, the expression of E-cadherin decreased and Snail and MMP9 increased in A549 cells after ATP6V1B1-AS1 overexpression (P<0.05). After ATP6V1B1-AS1 deletion, the expression of E-cadherin in SK-MS-1 cells increased, and Snail and MMP9 proteins decreased (P<0.05). The ceRNA network regulation map results showed that miR-520a-5p, miR-526b-3p, miR-4524a-3p, and miR-6730-5p may be the downstream genes of ATP6V1B1-AS1. Those genes regulated WNT3, PAICS, and other genes to help ATP6V1B1-AS1 promote non-small cell lung cancer proliferation and invasion. GO functional enrichment analysis showed that the downstream mRNA of ATP6V1B1-AS1 was related to glandular development and other functions, while KEGG functional enrichment analysis showed that ATP6V1B1-AS1 was related to purine metabolism and other functions. Conclusion ATP6V1B1-AS1 is signifiacntly expressed in non-small cell lung cancer cell lines and can increase non-small cell lung cancer proliferation and invasion. ATP6V1B1-AS1 may regulate the corresponding biological behavior through the ceRNA network.
Key words: non-small cell lung cancer, long non-coding RNA, ATP6V1B1-AS1, proliferation, invasion
PDF全文下载地址:

https://journal.cmu.edu.cn/CN/article/downloadArticleFile.do?attachType=PDF&id=3141