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蛴螬粗提物对人宫颈癌HeLa细胞诱导凋亡作用及其机制

Inducing Apoptosis Effects and Mechanism on HeLa Human Cervical Carcinoma Cells by Grub Extract

  • 摘要: 目的 研究蛴螬粗提物对人宫颈癌HeLa细胞诱导凋亡作用,并探讨其作用机制。方法 采用流式细胞仪检测HeLa细胞的细胞周期分布相和细胞凋亡率;末端脱氧核苷酸转移酶(TdT)介导的脱氧核苷酸缺口末端标记(TdT-mediated dUTP nick end labeling)TUNEL法检测凋亡细胞;通过免疫细胞化学SP法测定凋亡相关基因产物p53、Fas、bcl-2、Bax蛋白的表达情况。结果 (1)蛴螬粗提物可以诱导HeLa细胞发生凋亡,加药组出现亚二倍体峰,并且G0/G1期细胞比例显著增加(P<0.01),而S期和G2/M期细胞比例则明显下降,将细胞阻滞在G0/G1期;(2)蛴螬粗提物作用细胞后凋亡细胞数目随时间延长而增多,凋亡指数与药物处理呈时间依赖性和剂量依赖性;(3)蛴螬粗提物作用后bcl-2、p53蛋白随提取物浓度表达均下降,Bax、Fas蛋白表达上升,四种蛋白表达各组间比较差异均有具有统计学意义(P<0.01)。结论 (1) 蛴螬粗提物对人宫颈癌HeLa细胞具有诱导凋亡作用;(2)蛴螬粗提取物诱导凋亡作用机制可能与细胞周期发生G0/G1期阻滞有关;并且通过下调bcl-2、p53蛋白表达,上调Bax、Fas蛋白表达,经由细胞凋亡的死亡受体通路和线粒体通路完成凋亡的启动和执行。

     

    Abstract: Objective To investigate the effect of the grub extract on proliferation and apoptosis of HeLa human cervical carcinoma cells in vitro and explore its probable molecular mechanisms. Methods The distribution of cells cycle and the apoptotic rate were observed by flow cytometry.Apoptotic cells were determined by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method.The p53,Fas, bcl-2,and Bax proteins were determined by immunohistochemical SP staining technique. Results (1) The grub petroleum extract was able to induce the apoptosis of HeLa cells;Typical sub-dipioid peaks were observed.The percentage of cells in G0/G1 phase were increased while the percentage of cells in S and G2/M phase were decreased after the grub extract treatment so the grub extract could arrest HeLa cells in G0/G1 phase; (2) The apoptosis of HeLa cells induced by the grub extract was in a concentration and time dependent manner.(3) After treated with the extract,bcl-2 and p53 protein expressions were decreased while Bax and Fas were increased; the expression of four kinds protein were significantly changed between different treatment groups (P<0.01). Conclusion (1)The grub petroleum ether extract has significant inhibition of proliferation and inducing apoptosis for HeLa human cervical carcinoma cell line in vitro.(2) Apoptosis of HeLa cells induced by the grub petroleum ether may be related with arresting cell cycle in G0/G1 phase.The mechanism of inducing apoptosis for HeLa might be related by down-regulation expression of bcl-2、p53 and up-regulation expression of Bax、Fas.It might be induce apoptosis in HeLa cells by the Fas-dependent pathway and mitochondria pathway.

     

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