摘要
目的:观察白藜芦醇是否通过Sirt3-SOD2-ROS途径诱导人卵巢癌SKOV3细胞凋亡。方法:体外培养SKOV3细胞,分别加入2.5、5、10、20、40和80 mg/L白藜芦醇,作用24 h后,MTT法检测白藜芦醇对SKOV3细胞活力的影响。将SKOV3细胞随机分为空白对照组、白藜芦醇(10 mg/L)组、白藜芦醇(20 mg/L)组和白藜芦醇(40 mg/L)组,分别处理24 h后,用Hoechst 33342染色,应用激光共聚焦显微镜观察细胞核变化;用荧光探针标记细胞内活性氧(ROS),应用激光共聚焦显微镜观察细胞内ROS变化;采用Western blot检测沉默交配型信息调节因子2同源蛋白3(Sirt3)、超氧化物歧化酶2(SOD2)、Bcl-2、Bax和cleaved caspase-3蛋白表达。结果:MTT结果显示,白藜芦醇(2.5、5、10、20、40和80 mg/L)作用SKOV3细胞24 h,SKOV3细胞活力显著降低,IC 50为(47.6±0.1)mg/L。应用激光共聚焦显微镜观察,与空白对照组相比,白藜芦醇各浓度组SKOV3细胞核内出现明显固缩,且着色增强现象。ROS荧光标记显示,与空白对照组相比,白藜芦醇各浓度组中ROS红色荧光数量显著减少。Western blot检测结果表明,与空白对照组相比,白藜芦醇各浓度组Sirt3、SOD2、Bax和cleaved caspase-3蛋白表达显著上升(P<0.05),而Bcl-2蛋白表达显著降低(P<0.05)。结论:白藜芦醇可以通过调控Sirt3-SOD2-ROS途径诱导SKOV3细胞凋亡。
AIM:To investigate whetier resveratrol induces apoptosis of human ovarian cancer SKOV3 cells through Sirt3-SOD2-ROS pathway.METHODS:SKOV3 cells were cultured in vitro and treated with resveratrol at 0,2.5,5,10,20,40 and 80 mg/L for 24 h.The inhibitory effect of resveratrol on the viability of SKOV3 cells was measured by MTT assay.SKOV3 cells were randomly divided into blank control group,10 mg/L resveratrol group,20 mg/L resveratrol group and 40 mg/L resveratrol group.After 24 h of treatment,Hoechst 33342 staining and confocal microscopy were used to observe the nuclear changes.The protein levels of silent mating type information regulation 2 homolog 3(Sirt3),superoxide dismutase 2(SOD2),Bcl-2,Bax and cleaved caspase-3 were determined by Western blot.RESULTS:Treatment with resveratrol at 2.5,5,10,20,40 and 80 mg/L for 24 h significantly reduced the viability of SKOV3 cells.The observation by confocal microscopy showed that the nucleus of SKOV3 cells was markedly condensed and heavily stained with the increase in the concentration of resveratrol.Compared with blank control group,the red fluorescence intensity of ROS in different concentrations of resveratrol groups was significantly reduced.The results of Western blot showed that the protein levels of Sirt3,SOD2,Bax and cleaved caspase-3 in resveratrol groups were significantly higher than those in control group,while the protein expression of Bcl-2 was significantly lower than that in control group(P<0.05).CONCLUSION:Resveratrol induces apoptosis of SKOV3 cells by regulating Sirt3-SOD2-ROS pathway.
作者
李松岩
于洋
刘师兵
徐路
孙奇
徐冶
LI Song-yan;YU Yang;LIU Shi-bing;XU Lu;SUN Qi;XU Ye(School of Basic Medical Science,Jilin Medical University,Jilin 132013,China;Medical Research Laboratory,Jilin Medical University,Jilin 132013,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第2期229-233,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81372793)
吉林省教育“十三五”科学技术研究项目(No.JJKH20191064KJ)
