生长抑素类似物对胆管癌SK-ChA-1细胞株及其相关基因表达的影响

Effects of somatostatin analogues on the cell cycle and the related gene expression in human cholangiocarcinoma cell line SK-ChA-1

目的 探讨生长抑素类似物SMS2 0 1 995抑制人胆管癌增殖时对SK ChA 1细胞细胞周期及cyclinD1 和p16蛋白的影响。方法 采用血清饥饿法将SK ChA 1细胞同步化 ,同步化释放后SMS2 0 1 995处理 48h收取不同时相细胞 ,以流式细胞仪 (flowcytometer)分析细胞周期相分布 ;并同时用免疫组化和原位杂交检测cyclinD1 和p16基因表达水平。结果 SMS处理SK ChA 1细胞后 6、12h可抑制SK ChA 1细胞经血刺激由G0 G1 进入S期。CyclinD1 蛋白表达在SMS处理SK ChA 1细胞后 6h明显减弱。p16蛋白在细胞周期各时相呈稳定性低表达。SK ChA 1细胞cyclinD1 和p16mRNA各时相表达无明显变化。结论 SMS诱导的cyclinD1 蛋白量降低可能与SMS干扰细胞G1 S期转换 ,阻遏SK ChA

Objective To investigate the effects of somatostatin analogue (SMS201 995, SMS) on the cell cycle of human cholangiocarcinoma SK ChA 1 cells and cell cycle regulated gene cyclinD1 and p16. Methods SK ChA 1 cells were synchronized by serum starvation. Cells at different phases were harvested at 48 h after SMS treatment of the synchronized SK ChA 1 cells. The distribution of cell cycles was analyzed by flow cytometry. The expression levels of cyclinD1 and p16 gene were detected by SABC immunohistochemistry and in situ hybidization. Results SMS could significantly inhibit the poorly serum synchronized SK ChA 1 cells to reenter the cell cycle from G 0/G 1 phase at 6 h and 12 h ( P <0.05) by serum stimulation. SMS inhibited CyclinD1 protein expression significantly in the liberation of synchronized SK ChA 1 cells at 6 h, but p16 protein expression remained unchanged. No changes of CyclinD1 and p16 mRNA were found at different time points. Conclusion These findings suggest that the amounts of G0/G1 and S phase are dependent on the level of cyclinD1, and the down regulation of cyclinD1 in SK ChA 1 cells may be due to the anti proliferation of SMS.