选择性环氧合酶-2抑制剂联合奥曲肽诱导人胃癌细胞的凋亡

目的探讨选择性环氧合酶-2抑制剂NS-398与奥曲肽联合应用对人胃癌细胞株BGC-823生长、凋亡的影响。方法体外培养BGC-823细胞,分别用NS-398(100μmol/L)与奥曲肽(1μmol/L)单独及联合处理不同时间后,倒置显微镜观察细胞形态学变化;观察生长曲线的变化;流式细胞仪检测细胞凋亡率;实时定量(Real-time)PCR检测COX-2mRNA的表达;Western blot法检测Caspase-3蛋白表达。结果倒置显微镜下,对照组BGC-823细胞生长良好,药物处理后,细胞变小、变圆,悬浮,联合组细胞形态学改变显著强于单纯用药组;药物作用后,细胞生长受抑制,出现负增长,联合组作用明显强于单纯用药组;流式细胞仪检测表明联合用药组诱导BGC-823细胞的凋亡率明显高于单一用药组和对照组(P0.01);各处理组均使BGC-823细胞COX-2mRNA表达下调(P0.05);药物处理后细胞Caspase-3蛋白表达明显增加。结论 NS-398、奥曲肽联合可协同抑制BGC-823细胞生长、增殖,其机制可能与下调COX-2mRNA表达、诱导肿瘤细胞凋亡相关。

Combined effects of selective cyclooxygenase-2 inhibitor and octreotide induce apoptosis of human gastic cancer cells

Objective To study the effect of NS-398, a selective cyclooxygenase-2 inhibtor, combined with Octreotide on the growth and apoptosis of human gastic cancer cell line BGC-823. Methods The cells were divided into 4 groups： control group, simple NS-398（100μmol/L） group, simple Octreotide （1μmol/L） group and combi nation group（100μmol/L NS-398＋1μmol/L Octreotide）. ① Morphologic changes of BGC-823 cells were observed by inversion microscopy. ②Growth curve was observed. ③Apoptosis index of cells was meas ured by flow cytometryfFCM）. ④The expression of COX 2 gene was detected by RT PCR. ⑤ The ex pression of Caspase 3 was detected by Western blot. Results ① When viewed by light microscopy, normal BGC-823 cells grew well. Treated by drugs, cells became small, round and suspended, and the change of cell morphology in the combination group was more conspicuous than in the other trentment groups. ② When treated by drugs, cell growth was inhibited, with negative growth and this effect was more conspic uous in the combination group than in the other treatment. ③ By FCM, apoptosis in the combination group was significantly higher than groups（P〈0.01）. ④ RT PCR showed that the expression of COX-2gene was lowered in the other （P〈0.05）. ⑤ The expression of Caspase 3 protein increased significantly in treated groups compared with that of the control group（P〈0.05）. Conclusion The combination of NS 398 and Octreotide can synergistically inhibit growth and proliferation of BGC-823 cells, which is likely con nected with downregulating COX-2 genetic expression and inducing apoptosis of tumor cells.