人参皂甙Rg3对乳腺癌MCF-7线粒体凋亡途径的影响

The Effect of Ginsenoside Rg3 on Breast Cancer MCF-7 Mitochondrial Pathway of Apoptosis

目的探讨20(R)–人参皂甙Rg3(SPG-Rg3)对人乳腺癌MCF-7细胞的诱导凋亡作用及其可能机制。方法人乳腺癌细胞MCF-7细胞株分为空白对照组、实验对照组及SPG-Rg3多个浓度组。利用MTT法观察人参皂甙Rg3对MCF-7细胞生长的抑制作用,并计算出IC-50,进一步确定其有效浓度;流式细胞术检测人参皂甙Rg3作用后MCF-7细胞周期的变化;利用AnnexinV-EGFP/PI双染法,检测人参皂甙Rg3诱导MCF-7细胞凋亡情况;免疫细胞化学染色检测MCF-7细胞凋亡与Fas、FasL蛋白表达的关系。结果 SPG-Rg3作用48h的IC-50为244.54μg/mL。流式细胞仪检测Rg3使MCF-7的S期细胞比率明显增加(P<0.01),凋亡率明显增加(P<0.01)。免疫细胞化学显示Rg3能使MCF-7细胞胞浆内细胞色素C增加(P<0.01)。结论 SPG-Rg3能诱导乳腺癌MCF-7细胞凋亡,其机制可能与诱导线粒体释放细胞色素C有关。

Objective To study the mitochondrial apoptotic pathway induced by 20 （R）- ginsenoside Rg3 （SPG-Rg3） in human breast cancer MCF-7 cells and the mechanism of induction of apoptosis. Methods Human breast cancer cell line MCF-7 cells were divided into control group,experimental control group and SPG-Rg3 multiple concentrations. MTT was adopted to observe the restraining growth effect intervented by ginsenoside Rg3,and the half inhibitory concentration（IC-50） was calculated to further determine the effective concentration;Flow cytometry was adopted to detect MCF-7 cell cycle after the role of Ginsenoside Rg3;Annexin V- EGFP/PI double staining was adopted to detect of Ginsenoside Rg3 induced apoptosis of MCF-7 cells;Immunocytochemistry was used to detect the relationship between MCF-7 cell apoptosis and expression of cytochrome C protein. Results 20（R）-ginsenoside Rg3 on the growth of MCF-7 breast cancer cells significantly inhibited,and the inhibition rate increased with the increase of its concentration. 20（R） -ginsenoside Rg3 on MCF-7 breast cancer cells in IC-50 in 48h,respectively 244.54μg/mL;Flow cytometry results showed that： 300μg/mL 20（R）-the role of ginsenoside Rg3 in MCF-7 cells 48 hours after S phase cells increased significantly（P0.01）,G2-M phase cells decreased significantly （P 0.05）,MCF-7 cell apoptosis was significantly increased （P 0.01）;Cytochrome C protein immunocytochemistry showed that：In experimental groups,levels of cytosolic cytochrome C were significantly higher （P 0.01）. Conclusion SPG-Rg3 on MCF-7 breast cancer cells significantly inhibited;SPG-Rg3 can induce apoptosis in MCF-7 breast cancer. The occurrence mechanism of death may be induced by cytochrome C released from mitochondrial.