特异性siRNA沉默YAP1基因表达对胰腺癌细胞恶性生物学行为的影响

The effects of downregulation of YAP1 gene expression by siRNA on malignant biological behavior of pancreatic cancer cells

目的 利用特异性siRNA沉默YAP1基因表达,探讨YAP1的表达对体外胰腺癌细胞增殖、凋亡和侵袭作用的影响.方法 利用脂质体转染法将含有YAP1特异性siRNA的真核表达质粒稳定转染至人胰腺癌PANC-1细胞株;实时定量PCR和Western blot法检测PANC-1细胞中YAP1基因的表达情况;CCK-8法、流式细胞及Transwells侵袭实验分别检测调控YAP1的表达对体外胰腺癌细胞增殖、凋亡和侵袭作用的影响.结果 我们成功建立YAP1 siRNA稳定转染的胰腺癌PANC-1细胞株.经实时定量PCR和Western blot证实siRNA组较空载体组胰腺癌细胞YAP1 mRNA转录和蛋白表达水平明显降低.CCK-8检测发现siRNA组较空载体组胰腺癌细胞增殖能力显著下降（P＜0.05）;流式细胞检测提示siRNA组较空载体组胰腺癌细胞凋亡率明显增加（P＜0.05）;Transwells侵袭实验显示siRNA组的穿膜胰腺癌细胞数显著多于空载体组（P＜0.05）. 结论 特异性siRNA沉默YAP1基因表达能显著抑制胰腺癌细胞的增殖和促进凋亡,同时明显抑制胰腺癌细胞的体外侵袭能力.

Objective To investigate the effects of YAP1 knockdown by siRNA on pancreatic cancer cell proliferation,apoptosis and invasion.Methods We constructed YAP1 siRNA plasmids,and then stably transfected into PANC-1 cells.The mRNA and protein levels of YAP1 expression in the stabletransfected cells were detected using qRT-PCR and Western blot assay.CCK-8,Flow cytometry and Transwells assays were applied to observe pancreatic cancer cell proliferation,apoptosis and invasion.Results mRNA and protein levels of YAP1 expression in the siRNA group were significantly decreased compared with the control group.The proliferation of YAP1 siRNA cells decreased notably compared with the control group （P ＜ 0.05）,while the apoptotic rates increased （P ＜ 0.05）.The number of invasive PANC-1 cells in the siRNA group increased compared with the control group （P ＜ 0.05）.Conclusions Downregulation of YAP1 by siRNA inhibits pancreatic cancer cell proliferation,induces apoptosis and hinders its invasive ability in vitro.