[重楼总皂苷对人胃癌细胞MKN-45凋亡及Fas/FasL信号通路的影响]

[Effect of Paridis Rhizoma total saponins on apoptosis of human gastric cancer cell MKN-45 and Fas/FasL signaling pathway].

作者信息

Fang Hai-yan, Gong Xiao-yan, Hong Xing-hui, Hua Ming-liu, Huang Jin-ling

出版信息

Zhongguo Zhong Yao Za Zhi. 2015 Apr;40(7):1388-91.

Abstract

OBJECTIVE

The study aimed to test if Paridis Rhizoma total saponins (PRTS) could induce apoptosis of human gastric cancer cell MKN-45.

METHOD

Based on the previous researches, PRTS was set by different concentrations to treat human gastric cancer cell for 12 h (5, 10, 20 mg x L(-1)). Fluorescent staining methods were adopted to observe apoptotic morphological changes of MKN-45. The apoptosis rates were analyzed by flow cytometry with Annexin V-FITC/PI staining. The enzymatic activities of caspase-3 and caspase-8 were measured by ELISA. The protein levels of Fas and FasL were detected by Western blotting.

RESULT

Under a fluorescence microscope, MKN-45 treated by PRTS was seen typical apoptotic morphological features. PRTS significantly increased the rate of apoptosis. Compared with the control group, there exsited significant differences in apoptosis rate of PRTS concentration of 20 mg x L(-1) (P < 0.01); besides, the enzymatic activities of caspase-3 and caspase-8 were promoted obviously after the effect of PRTS on MKN-45 cells for 12 h (P < 0.01). The protein levels of Fas and FasL in the MKN-45 were upgraded significantly.

CONCLUSION

PRTS can induce apoptosis of human gastric cancer cell MKN-45 , which is concerned with caspase-3 and caspase-8 and upgraded Fas and FasL.

摘要

目的

本研究旨在检测重楼总皂苷（PRTS）是否能诱导人胃癌细胞MKN-45凋亡。

方法

根据前期研究，设置不同浓度的PRTS（5、10、20 mg·L⁻¹）处理人胃癌细胞12 h。采用荧光染色法观察MKN-45凋亡的形态学变化。用Annexin V-FITC/PI染色通过流式细胞术分析凋亡率。用ELISA法检测caspase-3和caspase-8的酶活性。用Western印迹法检测Fas和FasL的蛋白水平。

结果

在荧光显微镜下，PRTS处理的MKN-45呈现典型的凋亡形态学特征。PRTS显著提高了凋亡率。与对照组相比，20 mg·L⁻¹浓度的PRTS凋亡率存在显著差异（P < 0.01）；此外，PRTS作用于MKN-45细胞12 h后，caspase-3和caspase-8的酶活性明显升高（P < 0.01）。MKN-45中Fas和FasL的蛋白水平显著升高。