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AIM:To investigate the relationship betweenhepatocarcinogenesis and the expression of connexin32(cx32),connexin43(cx43)mRNAs and proteins in vitro.METHODS Gap junction genes cx32 and cx43 mRNA inhepatocellular carcinoma cell lines HHCC,SMMC-7721 andnormal liver cell line QZG were detected by in situhybridization(ISH)with digoxin-labeled cx32,and cx43cDNA probes.Expression of Cx32 and Cx43 proteins in thecell lines was revealed by indirect immuno-fluorescence andflow cytometry(FCM).RESULTS Blue positive hybridization signals of cx32 andcx43 mRNAs detected by ISH with cx32 and cx43 cDNAprobes respectively were located in cytoplasm of cells ofHHCC,SMMC-7721 and QZG.No significant difference ofeither cx32 mRNA or cx43 mRNA was tested among HHCC,SMMG-7721 and QZG(P=2.673,HHCC vs QZG;P=1.375,SMMC-7721 vs QZG).FCM assay showed that thepositive rates of Cx32 protein in HHCC,SMMC-7721 and QZGwere 0.7%,1.7% and 99.0%,and the positive rates of Cx43protein in HHCC,SMMC-7721 and QZG were 7.3%,26.5%and 99.1% respectively.Significant differences of both Cx32and Cx43 protein expression existed between hepatocellularcarcinoma cell lines and normal liver cell line(P=0.0069,HHCC vs QZG;P=0.0087,SMMC-7721 vs QZG).Moreover,the fluorescent intensities of Cx32 and Cx43proteins in HHCC,SMMC-7721 were lower than that in QZG.CONCLUSION Hepatocellular carcinoma cell lines HHCCand SMMC-7721 exhibited lower positive rates andfluorescent intensities of Cx32,Cx43 proteins compared withthat of normal liver cell line QZG,It is suggested that lowerexpression of both Cx32 and Cx43 proteins in hepatocellularcarcinoma cells could play pivotal roles in thehepatocarcinogenesis.Besides,genetic defects of cx32 andcx43 in post-translational processing should be considered.
AIM: To investigate the relationship between hepatocarcinogenesis and the expression of connexin 32 (cx32), connexin43 (cx43) mRNAs and proteins in vitro. METHODS Gap junction genes cx32 and cx43 mRNA in hepatocellular carcinoma cell lines HZCC, SMMC-7721 and normal liver cell lines QZG were detected by in situ hybridization (ISH) with digoxin-labeled cx32, and cx43 cDNA probes. Expression of Cx32 and Cx43 proteins in the cell lines was revealed by indirect immuno-fluorescence and flow cytometry (FCM) .RESULTS Blue positive hybridization signals of cx32 and cx43 mRNAs detected by ISH with cx32 and cx43 cDNAprobes were respectively located in cytoplasm of cells ofHHCC, SMMC-7721 and QZG.No significant difference of inherent cx32 mRNA or cx43 mRNA was detected among HHCC, SMMG-7721 and QZG (P = 2.673, HHCC vs QZG; P = 1.375, SMMC-7721 vs QZG) .FCM assay showed that thepositive rates of Cx32 protein in HHCC, SMMC-7721 and QZGwere 0.7%, 1.7% and 99.0%, and the positive rates of Cx43protein in HHCC, SMMC-7721 and QZG were 7.3%, 26.5% and 99.1% respectively. Bothificant of both Cx32 and Cx43 protein expression existed between hepatocellularcarcinoma cell lines and normal liver cell line (P = 0.0069, HHCC vs QZG; P = 0.0087, SMMC-7721 vs QZG) and Cx43proteins in HHCC, SMMC-7721 were lower than that in QZG.CONCLUSION Hepatocellular carcinoma cell lines HHCC and SMMC-7721 were checked for lower positive rates and fluorescence intensities of Cx32, Cx43 proteins compared with the shift of the normal liver cell line QZG, It is suggested that lowerexpression of both Cx32 and Cx43 proteins in hepatocellular carcinoma cells could play pivotal roles in the hepatocarcinogenesis. Besides, genetic defects of cx32 and cx43 in post-translational processing should be considered.