【摘要】 MST1是死亡受体信号通路Hippo通路的核心成员,其基因表达异常多见于结直肠癌、肝癌、胃癌等肿瘤。为了探讨MST1表达对人结直肠癌SW480细胞增殖与凋亡的影响及其发生机制,采用PolyJet TM介导pEGFP-N1-Mst1及LipofectamineTM2000介导Mst1特异性-siRNA转染至SW480细胞,分别建立高低表达Mst1的细胞模型。对不同分组的细胞增殖、凋亡及凋亡相关蛋白的表达进行检测。结果显示,Mst1高表达组细胞增殖抑制、凋亡率及凋亡相关蛋白的表达显著增高,反之,Mst1低表达组细胞增殖加快,细胞凋亡率及凋亡相关蛋白的表达显著降低。结果提示,Mst1的靶向调控能较好地控制结直肠癌细胞的增殖与凋亡,可作为人结直肠癌防治的新研究靶点。更多还原

【Abstract】 To investigate the effects of targeted regulation of Mst1 expression on the proliferation and apoptosis of SW480 colorectal cancer cells and to elucidate the mechanisms underlying these effects.PolyJet TMin vitro DNA transfection reagent was used to transfect pEGFP-N1-Mst1 into SW480colorectal cancer cells,and LipofectamineTM2000 was used to transfect Mst1-specific siRNA for the targeted silencing of Mst1.MTS assay was then performed to detect the survival rate of the cells,flow cytometry was used to determine apoptosis,and western blot were used to measure the protein levels of MST1,PUMA,P73,P53,YAP,and caspase-3.Compared with the control group,the pEGFP-Mst1 and pEGFP-Mst1+5-FU groups showed significantly higher rate of cell proliferation inhibition and apoptosis(P<0.05).The protein levels of MST1,Phospho-YAP1(Ser127),p73,p53,PUMA,and caspase-3 were significantly increased(P<0.05).In the Mst1-siRNA group,the apoptosis rate was significantly decreased(P<0.01),cell proliferation was accelerated,p73,p53,and PUMA were downregulated,and YAP were upregulated.Conclusions The targeted regulation of Mst1 expression significantly affected the proliferation and apoptosis of the SW480 cells.Thus,Mst1 has potential for use as a new target for the prevention and treatment of colorectal cancer.更多还原