摘要
目的:探讨下调microRNA-205(miR-205)表达对人子宫内膜样腺癌细胞系Ishika-wa增殖和侵袭的作用及可能的机制。方法:将miR-205抑制剂(miR-205 inhibitor)瞬时转染Ishikawa细胞,下调miR-205的表达;应用实时荧光定量PCR方法(qRT-PCR)检测miR-205的表达;噻唑蓝(MTT)法检测细胞增殖能力;Transwell小室法检测细胞侵袭能力。qRT-PCR和Western blot分别检测miR-205的预测靶基因ESRRG mRNA和蛋白表达水平。应用双荧光素酶分析方法鉴定miR-205和ESRRG 3'UTR的表达调节关系。结果:miR-205在Ishikawa细胞中呈高表达;转染miR-205 inhibitor的Ishikawa细胞中,miR-205表达降低了86.9%,细胞增殖和侵袭能力下降,同时细胞ESRRG mRNA和蛋白表达升高;miR-205通过直接与ESRRG mR-NA 3'UTR结合负调节其表达。结论:下调miR-205表达可抑制Ishikawa细胞增殖、侵袭能力;miR-205的生物效应可能与调节潜在抑癌基因ESRRG有关。
Objective：To investigate the effects of down-regulated expression of hsa-miR-205 on the proliferation and migration of Ishikawa cells and its mechanism.Methods：miR-205 inhibitor was transiently transfected into Ishikawa cells to down-regulate expression of miR-205.The expression of miR-205 was detected using quantitative reverse transcription polymerase chain reaction（qRT-PCR）.Cellular proliferation and migration abilities were analyzed by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay and transwell migration assay,respectively.qRT-PCR and Western blot were used to detect estrogen-related receptor gamma（ESRRG） mRNA and protein expression,respectively,which was a putative target gene of miR-205.Luciferase assays were used to confirm the interaction between miR-205 and ESRRG.Results：miR-205 was expressed at higher levels in Ishikawa cells compared to 293T cells with a low level of miR-205.The miR-205 expression in miR-205 inhibitors transfected cells decreased 86.9%.The celluar proliferation was inhibited significantly and the cell migration activity decreased,compared to control transfected cells.Moreover,ESRRG was confirmed as a novel downstream target of miR-205,and inhibition of miR-205 increased endogenous ESRRG mRNA and protein expression was in vitro.Conclusions：The miR-205 down-regulation can suppress the cellular proliferation and migration of Ishikawa cells by targeting and depressing the potential tumor suppressor ESRRG.
出处
《现代妇产科进展》
CSCD
2012年第7期520-524,共5页
Progress in Obstetrics and Gynecology
基金
国家自然科学基金资助项目(No:81072139)
