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目的 探讨Gastrin、CCK对胆管癌细胞bcl 2和bax基因表达的调控作用。方法 用免疫组织化学、原位杂交、流式细胞术、逆转录 聚合酶链反应 (RT PCR)等技术 ,研究了Gastrin 17肽 (G 17)和CCK 8S肽 (CCK 8Sulfated)及CCK A受体拮抗剂L364 ,718(L18)和Gastrin/CCK B受体拮抗剂L365 ,2 60 (L60 )对QBC939胆管癌细胞bcl 2和baxmRNA、蛋白含量的影响。结果 在正常情况下胆管癌细胞bcl 2和baxmRNA表达均为阳性 ,baxmRNA强于bcl 2mRNA ;G 17( 1× 10 -10 mol/L、48h)、CCK 8S( 1× 10 -10 mol/L)明显增强bcl 2mRNA表达 ,而对baxmRNA表达无影响 ;同时加入L18和 /或L60 ( 1× 10 -10 mol/L)可明显抑制G 17和CCK 8S对bcl 2mRNA的促表达作用。经G 17或CCK 8S( 1× 10 -8mol/L)作用 48h ,胆管癌细胞bcl 2蛋白表达明显增加 ,而bax蛋白表达无显著变化 ;L60或L18( 1× 10 -8mol/L)能明显抑制G 17和CCK 8S对bcl 2蛋白表达的促进作用。结论 Gastrin和CCK对bcl 2基因表达有上调作用 ,提示Gastrin和CCK对胆管癌细胞凋亡过程有抑制作用。
Objective To investigate the effects of Gastrin and CCK on the gene expression of bcl-2 and bax in cholangiocarcinoma cells. Methods The effects of Gastrin 17 peptide (G 17), CCK 8Sulfated (CCK 8Sulfated) and CCK A receptor were studied by immunohistochemistry, in situ hybridization, flow cytometry and reverse transcription polymerase chain reaction (RT PCR) Effects of antagonists L364, 718 (L18) and Gastrin / CCK B receptor antagonist L365, 2 60 (L60) on bcl 2, bax mRNA and protein contents in QBC939 cholangiocarcinoma cells. Results The expression of bcl 2 and bax mRNA in cholangiocarcinoma cells was normal and the bax mRNA was stronger than that of bcl 2 mRNA in normal condition. The expression of bcl 2 mRNA in G 17 (1 × 10 -10 mol / L, 48 h), CCK 8S (1 × 10 -10 mol / L) Significantly increased the expression of bcl 2 mRNA, but had no effect on the expression of bax mRNA. The addition of L18 and / or L60 (1 × 10 -10 mol / L) significantly inhibited the expression of bcl 2 mRNA by G 17 and CCK 8S. The expression of bcl 2 protein in cholangiocarcinoma cells was significantly increased after treated with G 17 or CCK 8S (1 × 10 -8 mol / L) for 48 hours, while the expression of bax protein did not change significantly. L60 or L18 (1 × 10 -8 mol / L) Inhibit the promotion of bcl 2 protein expression by G 17 and CCK 8S. Conclusion Gastrin and CCK upregulate bcl 2 gene expression, suggesting that Gastrin and CCK can inhibit the apoptosis of cholangiocarcinoma cells.