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半边旗5F对HepG2细胞的凋亡作用及机制

[Apoptosis effect and mechanism of 5F from Pteris semipinnata on HepG2 cells].

作者信息

Li Li, Liu Yi, Lv Ying-Nian, Wu Ke-Feng, Chen Gong, Liang Nian-Ci

机构信息

Guangdong Key Laboratory for Research and Development of Natural Drugs, Guangdong Medical College, Zhanjiang 524023, China.

出版信息

Zhong Yao Cai. 2010 Jan;33(1):77-80.

Abstract

OBJECTIVE

To investigate the effect of Pteris semipinnata L. (PsL) extract Ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid (5F)-on HepG2 cells and explore its potential mechanism.

METHODS

Cytotoxicity of 5F was studied in HepG2 cells treated with different doses of 5F (0 - 80 mg/L) for 24 h and cell viability was determined by MTT assay. To analyze apoptosis qualitatively, the Hoechst/PI assay was used. The level of Bax in mitochondria fraction of 5F-treated HepG2 cells was determined by western blotting. The levels of cyto-c and AIF in the cytosol were analyzed by western blotting.

RESULTS

The cytotoxicity of 5F on HepG2 cells was elevated with the increasing of 5F concentrations, as evidenced by the cell viability assay. The apoptotic cells characterized by condensed neclei were observed after the exposure of HepG2 cells to 5F. The level of Bax in mitochondria fraction of 5F-treated HepG2 cells increased. The levels of cyto-c and AIF in the cytosol of 5F-treated HepG2 cells increased.

CONCLUSION

5F mediated apoptosis involves mitochondria-dependent pathway and 5F might have a therapeutic value against human cancer cell lines and especially on hepatocellular carcinoma (HCC) cells.

摘要

目的

研究半边旗提取物Ent-11α-羟基-15-氧代-贝壳杉-16-烯-19-酸(5F)对HepG2细胞的作用并探讨其潜在机制。

方法

用不同剂量的5F(0 - 80 mg/L)处理HepG2细胞24小时,通过MTT法检测5F的细胞毒性并测定细胞活力。采用Hoechst/PI法进行凋亡的定性分析。通过蛋白质免疫印迹法测定5F处理的HepG2细胞线粒体部分中Bax的水平。通过蛋白质免疫印迹法分析细胞质中细胞色素c和凋亡诱导因子(AIF)的水平。

结果

细胞活力测定表明,随着5F浓度的增加,其对HepG2细胞的细胞毒性增强。HepG2细胞暴露于5F后,观察到细胞核浓缩的凋亡细胞。5F处理的HepG2细胞线粒体部分中Bax的水平升高。5F处理的HepG2细胞细胞质中细胞色素c和AIF的水平升高。

结论

5F介导的凋亡涉及线粒体依赖性途径,5F可能对人类癌细胞系尤其是肝癌(HCC)细胞具有治疗价值。

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