摘要：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0.51~8.13 μmol·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3.67~11.1 μmol·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 AG4 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. Results The IC50 value of A549 , BeL-7402 , BGC-823 , C6, EJ, HeLa, HepG2, MCF-7 and LS180 cells pro-cessed 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 mitochon-drial pathway.
