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目的 研究肝细胞肝癌和正常肝细胞间隙连接蛋白Cx32、Cx43酪氨酸磷酸化作用的信号转导机制 ,及其对肝癌的重要意义。方法 应用细胞培养 ,Westernblot分析技术 ,研究肝癌细胞系HHCC ,SMMC 772 1和正常肝细胞系QZG中 ,间隙连接蛋白Cx32、Cx43的表达及酪氨酸磷酸化作用。结果 Westernblot分析表明 ,Cx32蛋白在正常肝细胞系QZG细胞中高表达 ,但未出现酪氨酸磷酸化作用 ,Cx32蛋白在HHCC ,SMMC 772 1无明显表达及酪氨酸磷酸化作用。Cx43蛋白在QZG ,SMMC 772 1细胞一定水平的表达仅在SMMC 772 1细胞表现出酪氨酸磷酸化现象 ,在QZG细胞中无酪氨酸磷酸化作用。Cx43蛋白在HHCC细胞中无明显表达及酪氨酸磷酸化现象。结论 肝癌细胞Cx32、Cx43蛋白翻译后水平酪氨酸位点磷酸化修饰作用 ,是调控间隙连接通讯功能的关键 ,间隙连接基因connexin32 ,connexin43翻译后水平调节和信号转导机制异常可能与肝癌的发生密切相关。
Objective To study the signal transduction mechanism of Cx32 and Cx43 tyrosine phosphorylation in hepatocellular carcinoma and normal hepatocytes and their significance in hepatocellular carcinoma. Methods Cell culture and Western blot analysis were used to study the expression of connexinx, connexinx and tyrosine phosphorylation in hepatocellular carcinoma cell lines HHCC, SMMC 772 1 and normal liver cell line QZG. Results Western blot analysis showed that Cx32 protein was overexpressed in normal liver cell line QZG cells but no tyrosine phosphorylation was observed. Cx32 protein was not expressed in HHCC and SMMC772 1 cells and tyrosine phosphorylation was observed. Expression of Cx43 protein at QZG, SMMC 772 1 cells at a certain level showed tyrosine phosphorylation only in SMMC 772 1 cells and no tyrosine phosphorylation in QZG cells. Cx43 protein was not expressed in HHCC cells and tyrosine phosphorylation. Conclusions Phosphorylation of Cx32 and Cx43 at the level of tyrosine phosphorylation after translation in hepatocellular carcinoma cells is the key to the regulation of gap junctional communication. The abnormalities of connexin32 and connexin43 posttranslational regulation and signal transduction may be related to the occurrence of hepatocellular carcinoma closely related.