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首页> 外文期刊>World Journal of Gastroenterology >Signal transduction of gap junctional genes, connexin32, connexin43 in human hepatocarcinogenesis
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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 [Ca~(2+)]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 [Ca~(2+)]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 [Ca~(2+)]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。方法：采用疤痕上色和染料转移（SLDT）和路西法黄（LY）检测肝癌细胞HHCC，SMMC-7721和正常对照肝细胞QZG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。通过免疫印迹检查连接蛋白蛋白酪氨酸上的磷酸化。结果：SLDT显示QZG细胞的GJIC功能高于HHCC和SMMC-7721细胞。通过激光扫描共聚焦显微镜，QZG细胞系（108.37 nmol / L）中细胞内游离钙[Ca〜（2 +）] i的浓度比HHCC（35.13 nmol / L）和SMMC-7721（47.08 nmol）中的高得多。 / L）单元格。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。 SMMC-7721细胞显示磷酸化的酪氨酸Cx43蛋白。结论：人肝细胞癌的发生，发展与cx基因表达异常及其信号转导途径异常有关，如[Ca〜（2 +）] i减少，酪氨酸翻译后磷酸化等。 Cx蛋白导致GJIC的戏剧性破坏。

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