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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 role of cytoskeleton in the proliferation of human colorectal cancer cells and its possible mechanism. Methods: The expression of C-MUC in colorectal cancer HT-29, HCT-116, SW620 and SW480 cells was detected by RT-PCR and immunofluorescence respectively. MTT assay was used to detect the effect of blocker SecinH3 on the proliferation of HT-29 cells after interfering with the expression of mucin-2 or small interfering RNA (siRNA). The effect of SecinH3 blockade or ARNO-siRNA interference on the activation of epidermal growth factor receptor (EGFR) pathway molecules in HT-29 cells was detected by Western blotting. The expressions of insulin-like growth factor type I (Insulin-like growth factor type I receptor, IGF-IR) pathway molecule activation. Results: The four subtypes of CAGE were expressed in HT-29, HCT-116, SW620 and SW480 cells, and the expression of ARNO was higher. After treated with SecinH3, the growth of HT-29 cells was inhibited, The inhibitory rate of ARNO-siRNA interfered with ARNO protein was 72h, and the proliferation of HT-29 cells was inhibited after ARNO-siRNA was transfected into HT-29 cells for 48h The rate was (58.95 ± 3.42)%. The results of Western blotting showed that both SecinH3 and ARNO-siRNA could down-regulate the expression of ARNO protein in colorectal cancer cells, which affected EGFR signaling pathway in HT-29 cells and IGF-IR signaling pathway in HCT-116 cells Inhibition, the corresponding downstream signaling pathway factor also appeared significant down. CONCLUSION: E-cadherin is correlated with the activation of EGFR or IGF-IR signaling pathway, which may be a new target for the treatment of colorectal cancer.