基础医学与临床 ›› 2017, Vol. 37 ›› Issue (11): 1590-1595.

• 研究论文 • 上一篇    下一篇

miR-211通过靶向调控TFAM抑制人乳腺癌细胞系增殖

肖顺崇,罗汉传,覃俊仕   

  1. 广西医科大学第八附属医院 贵港市人民医院
  • 收稿日期:2016-10-08 修回日期:2016-12-30 出版日期:2017-11-05 发布日期:2017-11-01
  • 通讯作者: 罗汉传 E-mail:luohanchuan@126.com

miR-211 inhibits the proliferation of human breast cancer cell lines through targeting TFAM

  • Received:2016-10-08 Revised:2016-12-30 Online:2017-11-05 Published:2017-11-01

摘要: 目的 探讨miR-211靶向线粒体转录因子A(TFAM)对人乳腺癌细胞增殖的影响。方法 用miR-211及TFAM作为研究对象。首先,在乳腺癌细胞中转染miR-211 mimics或miR-211 抑制剂以实现miR-211过表达或miR-211沉默,并检测miR-211过表达或沉默时TFAM蛋白质的表达水平;其次,构建了在TFAM的5’端有或无6对碱基突变的荧光酶报告基因质粒(mut-TFAM/wt-TFAM),与miR-211 mimics或miR-211 抑制剂共转染后检测荧光酶活性变化;然后,构建pcDNA3.1/TFAM质粒,与miR-211 mimics或miR-211 抑制剂共转染后检测TFAM蛋白质表达水平变化;最后检测pcDNA3.1/TFAM和mimics NC/miR-211 mimics共转染后乳腺癌细胞增殖的增殖。结果 miR-211过表达抑制TFAM蛋白质表达(P<0.01),miR-211沉默促进TFAM蛋白质表达(P<0.01);miR-211可靶向结合TFAM调控其表达;pcDNA3.1/TFAM可实现TFAM过表达(mRNA P<0.01,蛋白质 P<0.01),并可恢复miR-211对TFAM的抑制作用;miR-211可抑制乳腺癌细胞的增殖和增殖(P<0.05),TFAM可促进乳腺癌细胞增殖增殖(P<0.01),TFAM可回复miR-211对乳腺癌细胞增殖增殖的抑制作用(P<0.05)。结论 miR-211靶向TFAM基因抑制人乳腺癌细胞的增殖。

关键词: miR-211, TFAM, 生长, 增殖, 人乳腺癌

Abstract: Objective The present study was aimed to investigate the role of miR-211/TFAM in the regulation of proliferation of breast cancer cells. Methods In the present study, we choose miR-211 and TFAM as the research object. First we transfected breast cancer cells with miR-211 mimics or miR-211 inhibitor to achieve miR-211 overexpression or miR-211 silencing, and detected the expression of miR-211 and the expression level of TFAM proteins in response to miR-211 overexpression or miR-211 silencing; secondly luciferase reporter gene plasmid with or without a six base pairs mutation in the 3’UTR of TFAM (mut-TFAM/wt-TFAM) were conducted and co-transfected with miR-211 mimics or miR-211 inhibitor, then the changes of the luciferase activity were detected; then pcDNA3.1/TFAM plasmid was constructed and co-transfected with miR-211 mimics or miR-211 inhibitor, TFAM protein expression level changes were determined in response to miR-211 overexpression or miR-211 silencing; lastly the cell proliferation was determined in response to mimics NC/miR-211 mimics and pcDNA3.1/TFAM co-transfection. Results Overexpression of miR-211 inhibits the expression of TFAM protein, miR-211 silencing promote TFAM protein expression; miR-211 can regulate the expression of TFAM by direct targeting; TFAM overexpression was achieved by pcDNA3.1/TFAM transfection, and TFAM overexpression can restore the inhibitory effect of miR-211 on TFAM; miR-211 can inhibit the growth and proliferation of breast cancer cells, TFAM can promote the growth and proliferation of breast cancer cells; TFAM can restore the inhibitory effect of miR-211 on growth and proliferation of breast cancer cells. Conclusion miR-211 regulates the growth of breast cancer cell through targeting HMGB.

Key words: miR-211, TFAM, growth, proliferation, human breast cancer