Mst1对结直肠癌SW480细胞增殖与凋亡的影响  

Effect of targeted regulation of Mst1 expression on the proliferation and apoptosis of SW480 colorectal cancer cells

作　　者：刘卓琦[1] 卢洪飞 杨晓红[1] 万慧芳[2] 余乐涵[2] 许宝华[3] 张霞丽[3] 万福生[1] 

机构地区：[1]南昌大学基础医学院生物化学与分子生物学教研室,江西南昌330006 [2]南昌大学医学实验教学中心,江西南昌330006 [3]南昌大学实验动物科学中心,江西南昌330006

出　　处：《南昌大学学报（理科版）》2015年第6期606-612,共7页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金资助项目(81360032)

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。To investigate the effects of targeted regulation of Mstl expression on the proliferation and apop- tosis of SW480 colorectal cancer cells and to elucidate the mechanisms underlying these effects. PolyJet^TM in vitro DNA transfection reagent was used to transfect pEGFP-N1-Mstl into 8W480 eolorectal cancer cells, and Lipofectamine^TM2000 was used to transfect Mstl-specific siRNA for the targeted silencing of Mstl. MTS assay was then performed to detect the survival rate of the cells, flow cytometry was used to deter- mine 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-Mstl and pEGFP-Mstl ＋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 （PM0. 05）. In the Mstl-siRNA group, the apoptosis rate was significantly decreased （PM0.01）, 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,Mstl has potential for use as a new target for the prevention and treatment of colorectal cancer.

关 键 词：Mst1 结直肠癌 细胞增殖 凋亡 基因表达 

分 类 号：R34[医药卫生—基础医学]