走马胎中皂苷成分AG4对MCF-7肿瘤细胞增殖的影响及机制研究

Antiproliferation activity of triterpenoid saponins AG4 from Ardisia Gigantifolia Stapf.on MCF-7 cells

目的研究走马胎皂苷成分AG4对不同肿瘤细胞株增殖的影响,检测其对MCF-7细胞凋亡、细胞周期、Caspase-3和Caspase-9的活性、以及SOD活性和GSH、MDA含量的影响。方法终浓度为0.51～8.13μmol.L-1的AG4作用于肿瘤细胞株24～72 h后,采用MTT比色法检测AG4对9种肿瘤细胞株增殖的影响;筛选出对AG4敏感的细胞株,Hoechst染色观察细胞形态学变化;采用流式细胞术检测细胞凋亡和周期变化;采用相应试剂盒分别进行SOD活性和GSH、MDA含量的测定;采用Caspase-3和Caspase-9活性检测试剂盒对Caspase-3和Caspase-9活性进行检测。结果AG4作用于A549、BeL-7402、BGC-823、C6、EJ、HeLa、HepG2、MCF-7、LS180细胞24 h、48 h和72 h的IC50为3.67～11.1μmol.L-1,其中MCF-7细胞株对AG4较为敏感,24 h、48 h和72 h的IC50值分别为(3.67±0.61)、(3.76±0.66)和(4.03±0.47)μmol.L-1。高、中两剂量的AG4作用MCF-7细胞24 h后,细胞凋亡形态明显,发生早期凋亡;细胞阻滞在S期;SOD活性和GSH含量降低,MDA含量明显升高;Caspase-3和Caspase-9的活性均明显高于正常对照组。结论AG4对MCF-7细胞的增殖抑制作用最为明显。AG4干预MCF-7细胞内的氧化还原系统、阻滞周期并通过激活线粒体凋亡信号转导通路来诱导细胞凋亡。

Aim To investigate the influence of AG4 derived from Ardisia gigantifolia Stapf. on various tumor cells, and to detect its impact on cell apoptosis, cell cycle, and activities of Caspase-3 and Caspase-9, as well as the activity of SOD, and contents of GSH, MDA. Methods Nine kinds of tumor cells influenced by 0. 51 -8.13 μmol.L-1 AG4 , 24 h-72 h were detected by MTT reduction assay; cells sensitive to AC,4 were sifted through and stained, and their morphological changes were observed under fluorescent inverted microscope; their apoptosis and cell cycle changes were detected by flow cytometry; the activity of SOD, and contents of GSH and MDA were assayed with corresponding kits respectively, and the activities of Caspase-3 and Caspase-9 were detected as well. Resuits The IC50 value of A549, BeL-7402, BGC-823, C6, EJ, HeLa, HepG2, MCF-7 and LS180 cells processed 24 h, 48 h, or 72 h with AG4 was 3.67 - 11.1 μmol.L-1, among which that of MCF-7 cells, the most sensitive cell to AG4, was （3.67 ±0.61 ）, （3.76 ±0.66 ） and （4. 03 ±0. 47 ）μmol.L-1, respectively. 24 h later, the apoptosis of MCF-7 cells processed by the high and middle dose of AG4 was significant; the obstruction of cell cycle was at the S phase; the activity of SOD and contents of GSH decreased; MDA increased significantly; and activities of Caspase-3 and Caspase-9 were much more active than those of the control group. Conclusions AG4 restricts proliferation of MCF-7 cells obviously. AG4 induces apoptosis of MCF-7 cells by interfering the intracellular balance of redox system, blocking cell and activating mitochondrial pathway.