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Signal transduction of gap junctional genes connexin32 connexin43 in human hepatocarcinogenesis

机译:间隙连接基因连接蛋白32连接蛋白43在人类肝癌发生中的信号转导

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摘要

AIM: To investigate gap junctional intercellular communication (GJIC) in hepatocellular carcinoma cell lines, and signal transduction mechanism of gap junction genes connexin32(CX32), connexin43(CX43) in human hepatocarcinogenesis.METHODS: Scarped loading and dye transfer (SLDT) was employed with Lucifer Yellow (LY) to detect GJIC function in hepatocellular carcinoma cell lines HHCC, SMMC-7721 and normal control liver cell line QZG. After Fluo-3AM loading, laser scanning confocal microscope (LSCM) was used to measure concentrations of intracellular calcium [Ca2+]i in the cells. The phosphorylation on tyrosine of connexin proteins was examined by immunoblot.RESULTS: SLDT showed that ability of GJIC function was higher in QZG cell than that in HHCC and SMMC-7721 cell lines. By laser scanning confocal microscopy, concentrations of intracellular free calcium [Ca2+]i was much higher in QZG cell line (108.37 nmol/L) than those in HHCC (35.13 nmol/L) and SMMC-7721 (47.08 nmol/L) cells. Western blot suggested that only QZG cells had unphosphorylated tyrosine in CX32 protein of 32 ku and CX43 protein of 43 ku; SMMC-7721 cells showed phosphorylated tyrosine CX43 protein.CONCLUSION: The results indicated that carcinogenesis and development of human hepatocellular carcinoma related with the abnormal expression of CX genes and disorder of its signal transduction pathway, such as decrease of [Ca2+]i, post-translation phosphorylation on tyrosine of CX proteins which led to a dramatic disruption of GJIC.
机译:目的:探讨肝癌细胞系中的间隙连接细胞间通讯(GJIC),以及间隙连接基因connexin32(CX32),connexin43(CX43)在人类肝癌发生中的信号转导机制。和Lucifer Yellow(LY)一起检测肝癌细胞HHCC,SMMC-7721和正常对照肝细胞QZG中的GJIC功能。加载Fluo-3AM后,使用激光扫描共聚焦显微镜(LSCM)来测量细胞中细胞内钙[Ca 2 + ] i的浓度。结果:SLDT显示,QZG细胞的GJIC功能能力比HHCC和SMMC-7721细胞的GJIC功能高。通过激光共聚焦显微镜观察,QZG细胞系(108.37 nmol / L)中细胞内游离钙[Ca 2 + ] i的浓度远高于HHCC(35.13 nmol / L)和SMMC- 7721(47.08 nmol / L)细胞。蛋白质印迹表明,仅QZG细胞的CX32蛋白32 ku和CX43蛋白43 ku中具有未磷酸化的酪氨酸。结论:SMMC-7721细胞具有酪氨酸磷酸化的CX43蛋白。结论:人肝癌的发生,发展与CX基因的异常表达及其信号转导途径的异常有关,例如[Ca 2+ ] i,CX蛋白的酪氨酸翻译后磷酸化导致GJIC急剧破坏。

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