Gastrin、CCK对胆管癌细胞bcl-2、baX基因表达的调控

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Gastrin17肽(G-17)和CCK-8S肽(CCK-8 Sulfated)及CCK-A受体拮抗剂L364,718(L18)和Gastrin/CCK-B受体拮抗剂L365,260(L60)对QBC939胆管癌细胞bcl-2和bax mRNA、蛋白含量的影响.结果在正常情况下胆管癌细胞bcl-2和bax mRNA表达均为阳性,bax mRNA强于bcl-2 mRNA;G-17(1×10-10 mol/L、48 h)、CCK-8S(1×10-10mol/L)明显增强bcl-2 mRNA表达,而对bax mRNA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(1×10-8 mol/L)作用48 h,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(1×10-8 mol/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.

The modulation of Gastrin and CCK on gene expression of bcl-2 and bax of bile duct cancer cells

Objective To investigate the modulatory e ffect of Gastrin and cholecystokinin (CCK) on the expression of bile uct cancer cells bc1-2 and bax genes.Methods Technologies of immunohistochemistry,hybridizati on in situ,flow cytometry (FCM),RT-PCR were used to study the influence of Ga strin-17 (G-17) and CCK-8S (CCK-8 Sulfated ),and CCK-A receptor antagonism L364,718 (L18),Gastrin/CCK-B receptor antagonism L365,260 (L60) on the cont ents of mRNA and protein in QBC939 bile duct cancer cells bc1-2 and bax.Results The expression of mRNA of bcl-2 and bax in the b ile duct cancer cell was positive under normal condition with the expression of bax mRNA stronger than bcl-2mRNA; G-17 (10~(-10)mol/L) and CCK-8S (10 ~(-10)mol/L,48h) could clearly increased the expression of bcl-2 mRNA, but had no influence on the expression of bax mRNA.Addition of L18 (10~(-10) mol/L) and/or L60 (10~(-10)mol/L) could obviously restrain the expression of bcl-2mRNA boosted by G-17 or CCK-8S.After incubation of CCK-8S (10~(-8 )mol/L) or G-17 (10~(-8)mol/L) for 48h,the expression of protein of bcl-2 in the bile duct cancer cell was remarkably increased,while there was no obvious change in the expression of bax albumen.Addition of L60 (10~(-8)mo l/L) or L18 (10~(-8)mol/L) could obv iously restrain the expression of bcl-2 protein boosted by G-17 and CCK-8S.Conclusion Gastrin and CCK have the modulatory function t o the genetic expression of bc1-2,indicating that Gastrin and CCK had the func tion of restraining apoptosis of bile duct cancer cells.