目的 研究香加皮水提取物(CPE)诱导人胃癌细胞BGC-823凋亡及其作用机制.方法 采用Giemsa染色观察细胞凋亡形态学变化;电子显微镜观察凋亡细胞的超微结构变化;流式细胞术和琼脂糖凝胶电泳方法检测BGC-823细胞凋亡率、细胞周期和细胞凋亡的DNA水平变化;RT-PCR方法检测细胞凋亡相关基因bcl-2、 bax和survivin mRNA表达水平变化;免疫细胞化学方法检测bcl-2、bax和survivin蛋白表达的变化.结果经CPE作用后,人胃癌细胞BGC-823出现明显的细胞凋亡形态学变化及超微结构改变,细胞DNA琼脂糖凝胶电泳呈现梯形图.经250μg/mL CPE处理48 h后,多数BGC-823细胞被阻滞在G2/M期,而且细胞发生明显的凋亡变化,BGC-823细胞凋亡率可达18.9%.CPE可抑制BGC-823细胞bcl和survivin mRNA及蛋白的表达,促进bax mRNA及蛋白的表达.CPE可明显延长S₁₈₀荷瘤小鼠生存期,且具有剂量依赖性.结论CPE通过阻滞BGC-823细胞于G2/M期及诱导BGC-823细胞凋亡发挥抗肿瘤作用,其作用机制与抑制细胞的bcl-2和survivin基因mRNA及蛋白表达、促进bax基因和蛋白的表达有关.

Objective To study the inductive effect ofCortex Periplocae extract(CPE)on apoptosis of human gastric cancer cells BGC-823 and its mechanism.Methods The cell morphology and super-microstructural changes of apoptosis were analysed by Giemsa stain-ing and electric microscope,respectively.The BGC-823 apoptosis ratio,cell cycles,and changes of apoptosis in DNA level were studied by Flow Cytometry and agarose gel electrophoresis.The genes mRNA and protein expression of apoptosis-related genes bcl-2,bax,and survivin were studied by RT-PCR and immunology cell chemistry method.Results After treatment with CPE,BGC-823 cells showed some typical morphologic features and super-microstructural changes of apoptosis.DNA agarose gel electrophoresis showed characteristic"DNA ladder" pattern.Most BGC-823 cells were arrested at G2/M phase.Some typical subdiploid peaks before G0/G1 phase were observed.The apoptotic rate of BGC-823 was 18.9%after 250 tlg/mL CPE treated for 48 h.The gene mRNA and protein expression of bcl-2 and survivin were inhibi-ted by CPE,whereas that of bax was up-regulated.CPE could enhance the 1ife span of S₁₈₀ bearing mice in a dose-dependent manner.Conclusion CPE can inhibit the tumor growth by arresting the BGC-823 cell cycle at G2/M phase and inducing BGC-823 apoptosis.Its mechanism iS related to the inhibition on gene mRNA and protein expression of bcl-2 and survivin,and enhancement of those of bax.

国家自然科学基金资助项目(30371753);河北省自然科学基金资助项目(C2004000610)