微小RNA-202对A549细胞增殖和周期的作用及机制

doi:10.3969/j.issn.1004-616x.2017.06.010

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (6): 454-459.doi: 10.3969/j.issn.1004-616x.2017.06.010

微小RNA-202对A549细胞增殖和周期的作用及机制

彭慧, 马书梅, 洪伟伟, 隋静, 张艳秋, 梁戈玉

东南大学公共卫生学院, 环境医学工程教育部重点实验室, 江苏南京 210009

收稿日期:2017-07-07 修回日期:2017-11-12 出版日期:2017-11-30 发布日期:2017-11-30
通讯作者: 梁戈玉,E-mail:lianggeyu@163.com E-mail:lianggeyu@163.com
作者简介:彭慧,E-mail:penghui2920@163.com。
基金资助:
国家自然科学基金项目（81472939，81673132）；江苏省科技厅2015年社会发展项目（BE2015719）；南京市科技计划项目（201503006）

Regulatory mechanism of miR-202 on proliferation of A549 lung cancer cells

PENG Hui, MA Shumei, HONG Weiwei, SUI Jing, ZHANG Yanqiu, LIANG Geyu

Key Laboratory of Environmental Medicine Engineering of Ministry of Education, School of Public Health, Southeast University, Nanjing 210009, Jiangsu, China

Received:2017-07-07 Revised:2017-11-12 Online:2017-11-30 Published:2017-11-30

摘要/Abstract

摘要： 目的：探讨miRNA-202对肺癌细胞A549的调控作用和可能机制。方法：应用DIANA TOOLs及KEGG数据库对miR-202进行生物信息学分析，探讨其可能参与的信号通路和潜在靶基因。通过慢病毒转染，分别上调和下调A549细胞中miR-202的表达，应用MTT法和流式细胞技术检测miR-202对细胞增殖、周期和凋亡的影响。应用qPCR和Western blot技术分别检测miR-202对其预测靶基因PIK3R3的mRNA和蛋白表达水平的调控作用。结果：生物信息学分析显示，miR-202参与的信号通路可能通过对PIK3CA的靶向调控发挥作用。功能研究表明，与阴性对照组相比，上调miR-202表达可抑制A549细胞增殖（P < 0.05），引起细胞周期G1/S期阻滞（P < 0.05），对细胞凋亡无明显影响（P > 0.05）。靶基因研究结果显示，与阴性对照组相比，上调miR-202表达可引起PIK3CA蛋白表达量下降（P < 0.05），但其mRNA的表达无显著改变（P > 0.05）。结论：miR-202可能通过对PIK3CA的负向调控影响A549细胞的增殖和周期，本研究为深入了解miRNA在肺癌中的作用与机制提供了新的线索。

关键词: 肺癌, miR-202, 细胞增殖, 细胞周期, 调控机制

Abstract: OBJECTIVE:To investigate the regulatory mechanism on proliferation of miR-202 in A549 lung cancer cells. METHODS:Through the lentivirus transfection into A549 cells,up-and down-expression of miR-202 were investigated. DIANA-TOOLs and KEGG database were used to analyze the bioinformatic of miR-202 expression and to explore the signaling pathways that might be involved with potential target genes. Then,MTT assay and flow cytometry technology were applied to detect the influence of miR-202 on cell proliferation,apoptosis and cycle. RT-qPCR and Western blot methods were applied to detect the miR-202 regulation for its predicted target gene on both mRNA and protein levels. RESULTS:Bioinformatics analysis show that,miR-202 was involved in non-small cell lung cancer and other tumor-related pathways,and possibly in regulation of PIK3CA. Functional studies show that,compared with the negative control group,up-regulation of miR-202 inhibited cell proliferation (P < 0.05),caused cell cycle G1/S phase arrest (P < 0.05),but did not affect cell apoptosis (P > 0.05). Target gene study show that,compared with the control group,up-regulation of miR-202 reduced the expression of PIK3CA protein (P < 0.05),but had no effect on the expression of PIK3CA mRNA(P > 0.05). CONCLUSION:Expression of miR-202 influenced the proliferation and cycle of A549 cells through the negative regulation of PIK3CA and the study provides a new clue to the role and mechanism of miRNA in lung cancer.

Key words: Lung cancer, miR-202, cell proliferation, cell cycle, regulatory mechanism

中图分类号:

R730.2

彭慧, 马书梅, 洪伟伟, 隋静, 张艳秋, 梁戈玉. 微小RNA-202对A549细胞增殖和周期的作用及机制[J]. 癌变·畸变·突变, 2017, 29(6): 454-459.

PENG Hui, MA Shumei, HONG Weiwei, SUI Jing, ZHANG Yanqiu, LIANG Geyu. Regulatory mechanism of miR-202 on proliferation of A549 lung cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(6): 454-459.

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微小RNA-202对A549细胞增殖和周期的作用及机制

Regulatory mechanism of miR-202 on proliferation of A549 lung cancer cells

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