目的研究蛴螬石油醚提取物对人宫颈癌HeLa细胞增殖和凋亡的影响,并探讨其作用机制。方法MTT法检测蛴螬石油醚提取物对体外培养的HeLa细胞株增殖抑制作用及细胞毒活性;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-mediateddUTPnickendlabeling,TUNEL)法检测细胞凋亡;通过免疫细胞化学S-P法测定凋亡相关基因产物Bcl-2、Bax蛋白与凋亡相关蛋白caspase-8和caspase-3的表达情况。结果蛴螬石油醚提取物对HeLa细胞的增殖抑制作用呈剂量依赖性和时间依赖性(P<0.01)。蛴螬石油醚提取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。蛴螬石油醚提取物作用后Bcl-2蛋白表达均下降,Bax、caspase-8和caspase-3表达上升,4种蛋白表达各组间比较均差异显著(P<0.01)。结论蛴螬石油醚提取物对HeLa细胞具有抑制增殖及诱导凋亡作用。蛴螬石油醚提取物对HeLa细胞抑制增殖作用(50μg/mL作用48h之前)主要以诱导凋亡为主。蛴螬石油醚提取物诱导凋亡作用可能与下调Bcl-2表达,上调Bax、caspase-8和caspase-3表达有关。

Objective To investigate the effect of petroleum ether extract from the grub of Holotrichia diomphalia on proliferation and apoptosis of human cervical carcinoma HeLa cells in vitro and explore its probable molecular mechanisms. Methods The growth inhibition of petroleum ether extract from the grub of H. diomphalia on HeLa cells was measured with MTT assay. Apoptosis cells were determined by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. The Bcl-2, Bax, caspase-8, and caspase-3 protein expressions were determined by immunohistochemical S-P staining technique. Results Petroleum ether extract from the grub of H. diomphalia significantly inhibited the proliferation of HeLa cells in a dose-and time-dependent manner (P0.01). The effect of petroleum ether extract from the grub of H. diomphalia increased HeLa apoptosis numbers greatly with the time lasting. The apoptosis of HeLa could be induced by the grub extract in a dose-and time-dependent manner. After treated with the extract, the Bcl-2 expression was decreased, while the Bax, caspase-8, and caspase-3 expressions were increased; the expressions of the four kinds of protein were significantly changed among the different treatings with the grub extract (P0.01). Conclusion Petroleum ether extract from the grub of H. diomphalia could significantly inhibit the proliferation and induce the apoptosis of HeLa cell in vitro. Before treating 48 h on HeLa cells with the grub extract (50 μg/mL), the inhibitory effect on proliferation and its cytotoxicity was mainly completed by the induction of apoptosis. The mechanism of the grub petroleum ether extract inducing apoptosis for HeLa might be related to down-regulation of Bcl-2 expression and up-regulation of Bax, caspase-8, and caspase-3 expressions.

国家自然科学基金资助项目(30460047)