MACC1基因影响宣威肺腺癌细胞XWLC-05促血管生成的机制研究  

Effect of MACC1 inhibition by small interference RNA on Xuanwei lung cancer cell XWLC-05 angiogenesis and its mechanism

作　　者：袁兵[1,2] 汤琦[1] 张云辉[2] 廖凤[2] 谭燕[2] 黄云超[1] 

机构地区：[1]昆明医科大学第三附属医院,昆明650118 [2]云南省第一人民医院,昆明650031

出　　处：《军事医学》2014年第10期803-806,共4页Military Medical Sciences

基　　金：云南省科技厅应用基础研究联合基金资助项目(2011FB208)

摘　　要：目的探讨结肠癌转移相关基因1(metastasis-associated in colon cancer 1,MACC1)影响宣威肺腺癌细胞XWLC-05促进血管生成的可能分子机制。方法将MACC1小干扰RNA(siRNA)-3转染XWLC-05宣威肺腺癌细胞株,采用Western印迹法检测细胞中MET、MEK/p-MEK、ERK/p-ERK、AKT/p-AKT蛋白的表达;ELISA法检测细胞中血管内皮细胞生长因子(VEGF)、IL-8和碱性成纤维细胞生长因子(bFGF)的表达。结果 Western印迹结果显示,MACC1 siRNA-3转染细胞中的MET、p-MEK和p-ERK表达水平显著降低,但总MEK和ERK蛋白表达无明显差异。同时,AKT及其磷酸化蛋白水平均未受影响。ELISA结果显示,MACC1 siRNA-3转染细胞48和72 h后的VEGF、bFGF和IL-8细胞因子的表达水平明显受到抑制。结论推测MACC1基因通过抑制XWLC-05宣威肺腺癌细胞中的HGF/c-MET和MEK/ERK信号通路,减少细胞中VEGF、bFGF和IL-8细胞因子的分泌,最终导致肺癌血管生成。Objective To study the effect of metastasis-associated in colon cancer 1 （ MACC1 ） inhibition by small interference RNA（ siRNA） on the formation of tumor cells that promote blood vessels, and to explore its possible molecular mechanism.Methods Synthesized MACC1 specific siRNA was transfected into human lung cancer cell lines XWLC-05. Then, the inhibitory effect of siRNA on the protein level of MET, MEK/p-MEK, ERK/p-ERK, and AKT/p-AKT was detected using Western blotting while the change in VEGF, bFGF and IL-8 protein level was detected using ELISA method. Results Western blotting results indicated that the expression level of MET, p-MEK and p-ERK was significantly decreased in the XWLC-05 cells transfected by MACC1 specific siRNA compared with control and nonsense siRNA groups, but the expression of total protein MEK and ERK did not change.Furthermore, the level of AKT and phosphorylated protein was not affected.ELISA results suggested that the secretion level of VEGF, bFGF and IL-8 was significantly suppressed at 48 h or 72 h compared with control and nonsense siRNA groups.Conclusion MACC1 may reduce the secretion levels of VEGF, bFGF and IL-8 via HGF/c-Met and MEK/ERK signal transduction pathways, and regulate angiogenesis of lung cancer.

关 键 词：结肠癌转移相关基因1 小干扰RNA MEK/ERK AKT 细胞因子 

分 类 号：R734.2[医药卫生—肿瘤]