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目的通过检测胃癌细胞系SGC7901/VCR、SGC7901和BGC-823,以及永生化胃上皮细胞系GES中B7-H1的表达,探讨B7-H1与胃癌的发生及多药耐药(MDR)的关系。方法上述细胞系培养于含10%灭活胎牛血清的RPMI-1640培养基中,利用RT-PCR技术、细胞免疫化学染色以及细胞免疫荧光染色技术与流式细胞术检测4种细胞系中B7-H1 mRNA与B7-H1蛋白的表达情况,并比较其在4种细胞系中的表达强度。结果B7-H1 mRNA在4种细胞系中均有表达,表达强度按照GES-1、BGC-823、SGC-7901、SGC-7901/VCR的顺序递增;细胞免疫化学染色与免疫荧光染色表明,B7-H1蛋白主要表达在上述细胞的细胞膜和少量细胞浆中,结合流式细胞术检测进一步证实,B7-H1蛋白在4种细胞系中的表达结果与mRNA的表达一致。结论B7-H1表达在胃上皮细胞上,可能通过抑制机体免疫反应,促进胃癌细胞生长及MDR基因表达。
Objective To investigate the relationship between B7-H1 and gastric carcinogenesis and multidrug resistance (MDR) by detecting the expression of B7-H1 in gastric cancer cell lines SGC7901 / VCR, SGC7901 and BGC-823 and immortalized gastric epithelial cell line GES. Methods The above cell lines were cultured in RPMI-1640 medium containing 10% inactivated fetal bovine serum. The expression of B7 in 4 cell lines was detected by RT-PCR, immunocytochemistry, immunofluorescence staining and flow cytometry -H1 mRNA and B7-H1 protein expression, and compare their expression intensity in four cell lines. Results B7-H1 mRNA was expressed in all four cell lines. The expression of B7-H1 mRNA increased in the order of GES-1, BGC-823, SGC-7901 and SGC-7901 / VCR. Immunocytochemistry and immunofluorescence staining showed that B7- -H1 protein was mainly expressed in the cell membrane and a small amount of cytoplasm. Combined with flow cytometry, the results of B7-H1 protein expression in four cell lines were consistent with the mRNA expression. Conclusion The expression of B7-H1 on gastric epithelial cells may promote the growth of gastric cancer cells and the expression of MDR gene by inhibiting the immune response.