杨梅素对人卵巢癌SKOV3细胞的凋亡诱导作用

发布时间：2018-10-19 07:21

【摘要】：目的:观察杨梅素对人卵巢癌SKOV3细胞的生长抑制及凋亡诱导作用,并探讨其可能机制。方法:体外培养人卵巢癌SKOV3细胞,按杨梅素给药浓度将细胞分为对照组和低、中、高浓度杨梅素组,杨梅素给药浓度分别为0、20、40和60mg·L-1。MTT法检测各组SKOV3细胞存活率;激光共聚焦显微镜观察各组SKOV3细胞核形态变化并计算异常细胞核比率;免疫荧光法检测各组SKOV3细胞中凋亡相关蛋白-活化半胱氨酸蛋白酶3(active Caspase-3)的表达;Western blotting法检测各组SKOV3细胞中DNA损伤相关蛋白γ-H2AX的表达水平。结果:与对照组比较,低、中和高浓度杨梅素组SKOV3细胞存活率明显下降(t=-12.35,t=-11.42,t=-10.21,P0.05);激光共聚焦显微镜下各浓度杨梅素组细胞出现核皱缩等凋亡形态学改变,且细胞中异常细胞核比率高于对照组(t=6.87,t=9.35,t=7.22,P0.05),高浓度杨梅素组异常细胞核比率高于低、中浓度杨梅素组(t=9.55,t=7.25,P0.05);随着杨梅素给药浓度的增加,SKOV3细胞中active Caspase-3的表达水平有升高趋势,各浓度杨梅素组SKOV3细胞中γ-H2AX的表达水平高于对照组(t=5.41,t=3.21,t=6.58,P0.05)。结论:杨梅素可抑制人卵巢癌SKOV3细胞增殖并诱导其凋亡,DNA双链断裂可能是其促凋亡的机制之一。
[Abstract]:Aim: to investigate the effect of myricetin on the growth inhibition and apoptosis induction of human ovarian cancer SKOV3 cells and its possible mechanism. Methods: human ovarian cancer SKOV3 cells were cultured in vitro. The cells were divided into two groups according to the concentration of myricetin: control group and low concentration group. The survival rate of SKOV3 cells was detected by 60mg L-1.MTT method. The morphologic changes of SKOV3 nuclei were observed by laser confocal microscope and the abnormal nuclear ratio was calculated. The expression of apoptosis-related protein-activated cysteine protease 3 (active Caspase-3) in SKOV3 cells was detected by immunofluorescence assay. The expression of DNA damage associated protein 纬 -H2AX in SKOV3 cells was detected by; Western blotting method. Results: compared with the control group, the survival rate of SKOV3 cells in the myricetin group was significantly lower than that in the control group (P 0.05), and the apoptotic morphologic changes such as nuclear shrinkage were observed in the myricetin group with laser confocal microscope, and the survival rate of the cells in the myricetin group was significantly lower than that in the control group (P 0.05). The percentage of abnormal nuclei in the cells was higher than that in the control group (t = 6.87), and the ratio of abnormal nuclei in the high concentration group was higher than that in the control group (P 0.05). The expression of active Caspase-3 in SKOV3 cells showed a tendency to increase with the increase of the administration concentration of myricetin, and the ratio of abnormal nuclei in the medium concentration group was higher than that in the control group. The expression level of 纬-H2AX in SKOV3 cells of myricetin group was higher than that in control group. Conclusion: myricetin can inhibit the proliferation and induce apoptosis of human ovarian cancer SKOV3 cells, and DNA double strand break may be one of its mechanisms.
【作者单位】：吉林医药学院基础医学院实验中心;吉林医药学院检验学院;吉林大学基础医学院病理生理学系;吉林省吉林中西医结合医院体检中心;吉林医药学院临床医学院;
【基金】：国家自然科学基金面上项目资助课题(81372793) 吉林省教育厅十二五科技项目资助课题(2013361) 吉林省大学生创新创业项目资助课题(20141027)
【分类号】：R737.31

【参考文献】