沉默Talin-1基因对前列腺癌PC-3细胞株上皮间质转化及侵袭能力的影响

Effects of the Talin-1 gene silencing on epithelial-mesenchymal transition and invasion of prostate cancer PC-3 cells

目的探讨沉默Talin-1基因对前列腺癌PC-3细胞上皮间质转化及侵袭能力的影响。方法RNAi法靶向沉默前列腺癌PC-3细胞Talin-1基因，选取PC-3空白对照组，PC-3加入空质粒（plsmifl）组、基因沉默组、基因沉默后加入转化生长因子p1（TGF-β1）组，PC-3加入TGF—p1组体外培养。MTT法检测各组细胞增殖能力，Transwell小室侵袭和迁移实验检测侵袭及迁移能力，Western-blot技术检测E—cadherin以及Vimentin蛋白表达情况。结果PC一3与PC-3＋plsmid两组增殖、侵袭和迁移能力以及E-cadherin和Vimentin蛋白表达水平未见明显差异（P〉0．05）；沉默Talin-1基因后，PC-3＋TGF-β1、PC-3、PC-3＋TGF—B1＋RNAi、PC-3＋RNAi中增殖、侵袭和迁移能力以及Vimentin蛋白表达水平依次递减，而E-cadherin蛋白表达水平依次递增（F-58．683、28．972、20．314、37．541、145．925，均P〈0．05）。结论沉默Talin_1基因后前列腺癌PC-3细胞生长受抑制，并且侵袭和迁移能力下降。Talin-1在前列腺癌侵袭和转移中发挥重要作用，其机制可能是通过升高Vimentin蛋白表达及降低E-cadherin蛋白表达从而诱发上皮间质表型转化而实现。

Objective To evaluate the effects of the Talin-1 gene silencing on invasion of prostate cancer PC-3 cells. Methods RNA interference（RNAi）was used to silence Talin-1 gene in prostate cancer PC-3 cells. The PC-3 cells were divided into blank control group,empty plasmid group, Talin-1 silencing group, Talin-1 silencing with adding of transforming growth factor 131（TGF-β1）group and TGF-β1 group,then cultured to logarithmic phase in vitro. MTT cell viability assay was used to detect cell viability rate,cell scratch assay and Transwell assays were used to assess the migration and invasion of the cells. Western blot analysis was used to measure the protein expression levels of E- cadherin and Vimentin. Results There were no significant differences in proliferation,invasion and migration ability as well as the expression levels of E-cadherin and Vimentin protein between blank control group and empty plasmid group （each P~ 0. 05）. The proliferation, invasion and migration ability were gradually decreased from TGF-β1 group to blank control group to Talin-l-silencing with TGF-131-added group, and to Talin-l-silencing group （ F = 58. 683,28. 972, and 20. 314, respectively, each P〈 0.05）. The protein expression levels of Vimentin were gradually decreased, whereas the protein expression levels of E-cadherin were gradually elevated from TGF-β1 group to blank control group to Talin-1 silencing with TGF-131 group and to Talin-1 silencing group（F： 37. 541 and 145. 925, respectively,each P〈 0.05）. Conclusions After silencing Talin-1 gene, the growth of prostate cancer PC-3 cells is inhibited, and accordingly, the invasion and migration ability are decreased. Talin-1 plays a significant role in invasion and migration of prostate cancer cells, and the possible mechanism is the inducing of epithelial-mesenchymal transition（ EMT） through elevating the expression of Vimentin and reducing the expression of E-cadherin.