miR-211通过靶向调控TFAM抑制人乳腺癌细胞系增殖

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pc DNA3.1/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pc DNA3.1/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(P0.01),miR-211沉默促进TFAM蛋白质表达(P0.01);miR-211可靶向结合TFAM调控其表达;pc DNA3.1/TFAM可实现TFAM过表达(mRNA P0.01,蛋白质P0.01),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(P0.05),TFAM可促进乳腺癌细胞增殖增殖(P0.01),TFAM可回复miR-211对乳腺癌细胞增殖增殖的抑制作用(P0.05)。结论 miR-211靶向TFAM基因抑制人乳腺癌细胞的增殖。

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

Objective To investigate the role of miR-211/TFAM in the regulation of proliferation of breast cancer cells .Methods In the present study , we transfected breast cancer cells with miR-211 mimics or miR-211 inhibitor to achieve miR-211 and detected the expression of miR-211 and the expression level of TFAM proteins in response to miR-211 overexpression or miR-211 silencing;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 change of the luciferase activity was 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 Overex-pression 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 over expression was achieved by pcDNA3.1/TFAM transfection , and TFAM overe xpression can restore the inhibitory effect of miR-211 on TFAM;miR-211 inhibited the growth and proliferation of breast cancer cells , TFAM can promote the growth and proliferation of breast cancer cells;TFAM restored the inhibitory effect of miR-211 on growth and proliferation of breast cancer cells .Conclusions miR-211 regulates the growth of breast cancer cell with targeting of HMGB .