胞黏蛋白对结直肠癌细胞增殖的影响及作用机制的初步探讨  

The effect of cytohesins on the proliferation of colorectal cancer cells and its related mechanism

作　　者：潘桃[1] 周俊[1] 孙俊峰[1] 王展怀[1] 付鲜花[1] 付志旋[1] 郑树[1] 张苏展[1] 

机构地区：[1]浙江大学附属第二医院肿瘤研究所,浙江省医学分子生物学重点实验室,教育部恶性肿瘤预警与干预重点实验室,杭州310009

出　　处：《肿瘤》2012年第5期320-326,共7页Tumor

摘　　要：目的:探讨胞黏蛋白在人结直肠癌细胞增殖中的作用及可能的作用机制。方法:分别采用RT-PCR法和免疫荧光法检测结直肠癌HT-29、HCT-116、SW620和SW480细胞中胞黏蛋白的表达情况。MTT法检测阻断剂SecinH3阻断胞黏蛋白,或小分子干扰RNA(small interfering RNA,siRNA)干扰胞黏蛋白-2(ARNO)表达后对HT-29细胞增殖的影响。蛋白质印迹法检测SecinH3阻断或ARNO-siRNA干扰对HT-29细胞中表皮生长因子受体(epidermal growth factor receptor,EGFR)通路分子激活情况的影响,对HCT-116细胞中胰岛素样生长因子I型受体(insulin-like growth factor type I receptor,IGF-IR)通路分子激活情况的影响。结果:胞黏蛋白的4个亚型在HT-29、HCT-116、SW620和SW480细胞中均有表达;其中以ARNO的表达量较高;SecinH3处理HT-29细胞后使细胞生长受到抑制,作用72h后细胞增殖抑制率达(57.22±1.01)%,ARNO-siRNA干扰ARNO蛋白的表达亦能使该肿瘤细胞的增殖受到抑制,ARNO-siRNA转染HT-29细胞48h后,其细胞增殖抑制率为(58.95±3.42)%。蛋白质印迹法检测结果提示,阻断剂SecinH3及ARNO-siRNA均能下调结直肠癌细胞中ARNO蛋白的表达,使HT-29细胞中EGFR信号通路、HCT-116细胞中的IGF-IR信号通路受到抑制,其相应的下游信号通路因子亦出现明显的下调。结论:胞黏蛋白与EGFR或IGF-IR信号通路的活化具有相关性,其有可能成为结直肠癌治疗的一个新的靶点。Objective：To investigate the effect of cytohesins on proliferation of colorectal cancer cells,and to explore its possible mechanism.Methods：The expressions of cytohesins in colorectal cancer HT-29,SW620,SW480 and HCT-116 cells were detected by RT-PCR and immunofluorescence.The proliferative activity of colorectal cancer HT-29 cells was detected by MTT method after SecinH3 or cytohesin-siRNA intervention.The activation of epidermal growth factor receptor（EGFR） signal pathway in HT-29 cells and the activation of insulin-like growth factor type Ⅰ receptor（IGF-IR） signal pathway in HCT-116 cells were detected by Western blotting.Results：The colorectal cancer HT-29,SW620,SW480 and HCT-116 cells expressed four homologous members of cytohesins,in which the cytohesin-2（ARNO） exerted a highest level.The proliferative-inhibiton rates of HT-29 cells blocked by SecinH3 for 72 h and interfered with ARNO-siRNA for 48 h were（57.22±1.01）% and（58.95±3.42）%,respectively;furthermore,the expression of ARNO protein was significantly decreased,and the EGFR and IGF-IR signaling pathways were both inhibited with down-regulation of factors in the corresponding downstream pathways.Conclusion：Cytohesins are associated with the activation of EGFR and IGF-IR signaling pathways,and the cytohesins maybe serve as a new molecular target in the treatment of colorectal cancer.

关 键 词：结直肠肿瘤 胞黏蛋白 表皮生长因子受体 胰岛素样生长因子I型受体 

分 类 号：R735.34[医药卫生—肿瘤]