生长抑素类似物诱导胆管癌细胞凋亡和bcl-2/bax表达的研究

Studies on apoptosis and bcl-2/bax gene expression induced by somatostatin analogue in human cholangiocarcinoma cell line

目的探讨生长抑素类似物SMS2 0 1 995 (SMS)对人胆管癌SK ChA 1细胞凋亡和凋亡调控基因bcl 2和bax的影响。方法用DNA凝胶电泳、流式细胞仪检测AnnexinV标记凋亡细胞、免疫组化和原位杂交方法研究SMS(10 0ng/ml)处理 6、12和 2 4h后人胆管癌SK ChA 1细胞凋亡和bcl 2 /bax表达的变化。结果SMS作用SK ChA 1细胞 6h对DNA无明显影响 ,作用 12和 2 4h后DNA凝胶电泳出现呈典型梯状条带。SMS作用 6、12和 2 4h后 ,AnnexinV标记的凋亡细胞分别为 (2 2±5 ) % ,(39± 7) %和 (5 8± 10 ) %。同时 ,SMS可使细胞bcl 2蛋白表达下降 ,使bax蛋白和mRNA表达升高。结论SMS能诱导SK ChA 1细胞发生凋亡 ,bax和bcl 2凋亡基因介导了SMS对SK ChA

Objective To study the effect of somatostatin analogue (SMS201-995,SMS) on apoptosis of human cholangiocarcinoma SK-ChA-1 cells and apoptosis-regulated gene bcl-2 and bax. Methods The changes of apoptosis and bcl-2/bax gene expression were detected by DNA gel electrophoresis, flowcytometric analysis and immunohistochemistry, in situ hybridization methods in cholangiocarcinoma SK-ChA-1 cells after treated with SMS(100?ng/ml) for 6, 12 and 24 hours.Results The characteristic ladder was not detected by agarose gel electrophoresis after treated with SMS(100?ng/ml) for 6 hours. When the cells were treated for 12 and 24 hours, the ladder of DNA electrophoresis was observed. While the cells were treated with SMS for 6, 12 and 24 hours, the percentage of annexin V-positive cells were (22±5)% ,(39±7)% and(58±10)%. Besides,SMS inhibited bcl-2 protein expression and increased bax protein and mRNA expression. Conclusion SMS can induce SK-ChA-1 cell apoptosis mainly with regulation of bcl-2 and bax gene expression.