不同恶化程度胃癌细胞系的细胞间隙连接通讯功能及Cx43表达的差异

DIFFERENT FUNCTIONS OF GAP JUNCTION INTERCELLULAR COMMUNICATION AND EXPRESSION OF GAP JUNCTION PROTEIN CONNEXIN 43 IN VARIOUS HUMAN GASTRIC CANCER CELL LINES WITH DIFFERENT MALIGNANT DEGREE

以人正常胃黏膜上皮细胞系GES-1和不同恶化程度的人胃癌细胞系(低恶化AGS细胞系、中恶化SGC-7901细胞系、高恶化BGC-823细胞系)为研究对象,采用激光共聚焦显微镜的荧光漂白恢复技术及划痕标记荧光染料示踪技术研究不同恶化程度的胃癌细胞系细胞间隙连接通讯(Gap junctional intercellular communication,GJIC)功能的差异;采用荧光定量PCR技术和间接免疫荧光技术检测间隙连接蛋白(Connexin43,Cx43)基因在mRNA和蛋白水平的表达差异,探求其与胃癌恶性程度的相关性.结果表明,在正常胃细胞系和低、中及高恶化胃腺癌细胞系中,细胞间的GJIC功能随着恶性程度的升高而减弱(AGS、SGC-7901)或消失(BGC-823),且Cx43蛋白及mRNA的表达量随着恶性程度的升高而下调(AGS、SGC-7901)或缺失(BGC-823).这提示胃癌恶性程度与胃癌细胞的GJIC功能显著相关(P<0.05),GJIC在胃癌发生中起重要作用.

To correlate function of gap junction intercellular communication（GJIC） and connexin43（Cx43） expression with carcinogenesis,progression and metastasis of gastric cells GES-1（human normal gastric mucosal cell line）,AGS（low malignant gastric cell line）,SGC-7901（middle malignant gastric cell line）,BGC-823（high malignant gastric cell line）,GJIC function and Cx43 expression at mRNA and protein levels were investigated using fluorescence recovery after photobleaching（FRAP） under laser scanning confocal microscope,scrape-loading dye transfer（SLDT）,real time quantitative PCR and indirect immunofluorescence assay.Data showed that with increasing malignancy,GJIC function become reduced or lost.GES-1 showed good GJIC function,AGS,SGC-7901 showed reduced GJIC function,but BGC-823 showed no GJIC function.Compared to GES-1,Cx43 expression（mRNA and protein） was downregulated in AGS and SGC-7901,but absent in BGC-823.This suggests that reduction in GJIC function and Cx43 expression was associated with occurrence,development and metastasis of gastric tumor.