生长抑素对胆囊癌细胞株的致凋亡研究

目的 通过前期实验发现,生长抑素(somatostatin,SST)作用于胆囊癌细胞株(GBC-SD)后,可显著增强阿霉素(doxorubicin,DOX)的杀伤能力[1].该实验旨在研究SST对GBC-SD细胞的诱导凋亡作用,并探讨其发生机制.方法 GBC-SD细胞分为4组:单独SST作用组、单独DoX作用组、联合用药组(SST预处理24 h后,再加入DOX)和对照组.药物作用24、36、48和60 h后,采用FITC-Annexin V/PI试验分别检测各组GBC-SD细胞的凋亡情况;另外运用Western印迹方法 检测SST作用24 h后,GBC-SD细胞内P53蛋白和Bax蛋白表达的变化.结果 SST分别作用24、36、48和60 h后,GBC-SD细胞没有出现明显的细胞凋亡,SST的致凋亡作用仅仅表现出微弱的时间依赖性;而在SST作用24 h后,GBC-SD细胞内的P53蛋白和Bax蛋白的表达都没有明显的变化(P>0.05).结论 SST不能诱导GBC-SD细胞产生明显的细胞凋亡,P53蛋白和Bax蛋白在其中发挥着重要作用.

Research of GBC-SD cell apoptosis induced by somatostatin

Objective In the previous research, after the treatment of somatostatin, the lethal effects of doxorubicin on gallbladder carcinoma were significantly enhanced. To identify the cytotoxici-ty of SST on GBC-SD cells, apoptosis index of GBC-SD cells after treatment of somatostatin or doxo-rubicin or co-use of somatostatin and doxorubicin was observed. Methods GBC-SD cells were divided into four groups: SST-alone-treated group, DOX-alone-treated group, co-treated group (co-treatment of SST and DOX) and the control group. In control group, the cells were cultivated by medium only. In SST-alone-treated group, the cells were cultivated by medium with SST in the concentration of 75 μg/ml. In DOX-alone-treated group, the cells were cultivated by medium with DOX in the concentra-tion of 5 μg/ml. In the co-treated group, cells were firstly cultivated by medium with 75 μg/ml SST for 24 h, followed by the addition of DOX in the concentration mentioned above. After treatment of the GBC-SD cells, apoptotic index was determined by FITC-Annexin V/PI assay at 24, 36, 48 and 60 h, respectively. Twenty-four hours after the treatment of SST, the expressions of P53 and Bax in GBC-SD cells was examined by western blot. Results After treatment of SST, no significant cell ap-optosis occurred. Only a weak time-dependent cytotoxicity of SST was observed. And 24 h after treat-ment of SST, the expression of P53 and Bax did not significantly varied either. Conclusion SST can not induce significant apoptosis of GBC-SD cells, in which P53 and Bax may play a critic role.