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目的:探讨醉茄素A体外诱导人胆囊癌GBC-SD细胞凋亡及其可能的分子机制。方法:以不同浓度醉茄素A处理人胆囊癌GBC-SD细胞后,CCK-8法检测GBC-SD细胞的增殖情况,Hoechst33258荧光染色和透射电子显微镜下观察GBC-SD细胞的凋亡形态和超微结构变化,FCM法检测GBC-SD细胞的凋亡率和周期分布,蛋白质印迹法检测GBC-SD细胞中多聚ADP核糖聚合酶[poly(ADP-ribose)polymerase,PARP]、cyclin D1、cyclin B1、细胞周期蛋白依赖性激酶1(cyclin-dependent kinase1,Cdk1)和p21蛋白的表达。结果:醉茄素A对GBC-SD细胞的增殖有明显的抑制作用(P<0.05),并且这种抑制作用呈浓度和时间依赖性。醉茄素A能诱导GBC-SD细胞出现典型的凋亡形态学改变及超微结构变化。与未加药的阴性对照组比较,醉茄素A作用后GBC-SD细胞的凋亡率明显增加(P<0.01),细胞阻滞于G2/M期,细胞周期相关蛋白cyclin D1和Cdk1的表达下调(P<0.05),cyclin B1和p21蛋白的表达上调(P<0.05),PARP蛋白剪切体增多(P<0.05)。结论:醉茄素A可能通过下调细胞周期相关蛋白cyclin D1、Cdk1的表达和上调cyclin B1、p21的表达,使GBC-SD细胞阻滞于G2/M期,进而抑制GBC-SD细胞的增殖并诱导其凋亡。
Objective: To investigate the in vitro apoptosis of GBC-SD cells induced by Saponin A and its possible molecular mechanism. Methods: The proliferation of GBC-SD cells was detected by CCK-8 assay with GBS-A cells treated with different concentrations of safranin A. The apoptosis morphology of GBC-SD cells was observed by Hoechst33258 fluorescence staining and transmission electron microscopy The apoptosis rate and cycle distribution of GBC-SD cells were detected by FCM and the expression of poly ADP-ribose polymerase (PARP), cyclin D1, cyclin B1, cyclin-dependent kinase 1 (Cdk1) and p21 protein. Results: Saponin A significantly inhibited the proliferation of GBC-SD cells (P <0.05), and the inhibitory effect was concentration-dependent and time-dependent. Saponin A can induce typical apoptosis morphological changes and ultrastructural changes in GBC-SD cells. Compared with the untreated negative control group, the apoptotic rate of GBC-SD cells increased significantly (P <0.01), the cells arrested in G2 / M phase, cyclin D1 and Cdk1 (P <0.05). The expressions of cyclin B1 and p21 protein were up-regulated (P <0.05) and the number of PARP protein spliced was increased (P <0.05). Conclusion: Saponin A inhibits the proliferation of GBC-SD cells by down-regulating the expressions of cyclin D1 and Cdk1 and up-regulating the expressions of cyclin B1 and p21 in GB / M phase Induce its apoptosis.