锰超氧化物歧化酶模拟化合物诱导人卵巢上皮性癌细胞株SKOV3凋亡的研究

Apoptosis of Human Ovarian Epithelial Cancer Cell Line SKOV3 Induced by Mimics of MnSOD

目的:研究锰超氧化物歧化酶模拟化合物(mimics of manganese superoxide dismutase,MnSODm)诱导人卵巢上皮性癌细胞株SKOV3凋亡的效应及机制。方法:取对数生长期的SKOV3细胞,用不同浓度的MnSODm(0、10、50μg/mL)处理0、24、48、72h,采用活细胞计数试剂盒(cell counting kit-8,CCK-8)检测细胞增殖能力与活性,采用PI单染和细胞形态学法观察细胞凋亡,采用荧光发光法检测caspase-3/7活性。结果:与空白对照组比较,MnSODm处理后SKOV3细胞的增殖受到抑制(P<0.01);10、50μg/mL MnSODm处理后,SKOV3细胞活性分别为(67.5±2.4)%及(16.6±0.9)%,与空白对照组的细胞活性[(100.0±1.7)%]比较,差异有统计学意义(P<0.01)。PI染色显示,MnSODm处理后的SKOV3细胞核边缘不规则,染色质凝集且着色较重,核固缩,核着色强阳性细胞及凋亡细胞较空白对照组明显增多。倒置显微镜显示,与空白对照组比较,MnSoDm处理后的SKOV3细胞伸展度降低,部分细胞体积变小、皱缩且折光性差,为细胞凋亡的形态改变。SKOV3细胞经10、50μg/mL MnSODm处理72h后,caspase-3/7的活性与空白对照组比较改变不明显(P>0.05)。结论:MnSODm能诱导SKOV3细胞凋亡,但并非通过caspase-3/7途径诱导。

Objective：To study the apoptosis-inducing effect of mimics of manganese superoxide dismutase（MnSODm） on human ovarian epithelial cancer cell line SKOV3 in vitro and its possible molecular mechanisms.Methods：SKOV3 cells in logarithmic growth phase were treated with different concentrations of MnSODm（0,10,50 μg/mL） for 0,24,48 h,72 h.Cell Couting Kit-8（CCK-8） was used to detect the cell proliferation and activity.The apoptosis of SKOV3 cells was observed with propidium iodide（PI） via inverted microscope.Caspase3/7 activity was detected by chemiluminescence method.Results： The proliferation and viability of cells have been obviously inhibited after treated by MnSODm,compared with those of the control group（P0.05）.PI staining revealed that the nucleus edges were irregular with condensed chromatin,shrinked nucleus and heavy coloring.The number of positive nuclear staining cells was much larger than that of the control group,and the apoptotic cells significantly increased.The inverted microscope displayed that the stretch of cells was reduced,and some cells were shrinked with weak refraction,all of which were the morphology changes of cell apoptosis.There were no significant differences in caspase3/7 activity between the two groups after 72h of drug treatment（P0.05）.Conclusions： MnSODm can induce SKOV3 cells apoptosis,but not through the way of caspase3/7.