白术多糖通过调控线粒体凋亡途径诱导人子宫内膜癌RL95-2细胞凋亡  

Polysaccharide of Atractylodes macrocephala induce apoptosis of human endometrial carcinoma RL95-2 cells by regulating mitochondrial apoptosis pathway

作　　者：杨宝芹[1] 刘高仁[2] 邵艳社 王自闯[3] YANG Baoqin;LIU Gaoren;SHAO Yanshe;WANG Zichuang(Department of Obstetrics and Gyneclogy,Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450003,China)

机构地区：[1]河南省中医院妇产科,郑州450003 [2]河南省中医院迎宾路院区急诊科,郑州450003 [3]河南省中医院病理科,郑州450003

出　　处：《中国免疫学杂志》2022年第12期1458-1462,共5页Chinese Journal of Immunology

摘　　要：目的:探讨白术多糖(PAM)对人子宫内膜癌RL95-2细胞凋亡的影响及其作用机制。方法:预实验选取梯度浓度(0、7.5、15、30、60、120、240 mg/ml)的PAM作用于RL95-2细胞,MTT比色法检测细胞增殖活性,Bliss法计算24 h、48 h和72 h的半数抑制浓度(IC50)。正式实验中将RL95-2细胞分为空白对照组和不同浓度(15、30、60 mg/ml)PAM处理组。共培养48 h后,Annexin V-FITC/PI双染法检测细胞凋亡;JC-1染色法检测细胞线粒体膜电位变化;Western blot检测细胞线粒体和细胞质中细胞色素C(Cyt C)蛋白水平差异;分光光度法检测细胞中半胱天冬酶9(Caspase9)活性;Western blot检测线粒体凋亡相关蛋白(Bcl-2、Bax和cleaved-Caspase3)表达水平。结果:PAM可降低RL95-2细胞的增殖率,与药物浓度和作用时间有关。与空白对照组相比,PAM处理组细胞凋亡率显著增加(均P<0.01);线粒体膜电位及Cyt C蛋白表达显著降低(均P<0.05),但细胞质中Cyt C蛋白表达显著增加(P<0.01);细胞中Caspase9活性显著增强(均P<0.05),Bcl-2蛋白水平显著降低(均P<0.05),而Bax和cleaved-Caspase3蛋白水平显著提高(均P<0.01)。结论:白术多糖可能通过激活线粒体凋亡途径促进RL95-2细胞凋亡。Objective:To investigate the effect of polysaccharide of Atractylodes macrocephala(PAM)on apoptosis of human endometrial carcinoma RL95-2 cells and its mechanism.Methods:PAM with gradient concentrations(0,7.5,15,30,60,120,240 mg/ml)were applied to RL95-2 cells in preliminary experiment.MTT colorimetric assay was used to detect proliferation activity of RL95-2 cells,and Bliss method was used to calculate 50%inhibitory concentrations(IC50)at 24 h,48 h and 72 h.In formal experiment,RL95-2 cells were divided into blank control group and PAM treated group with different concentrations(15,30,60 mg/ml).After co-culture for 48 h,Annexin V-FITC/PI double staining assay was used to detect cell apoptosis,and JC-1 staining assay was used to detect changes in mitochondrial membrane potential.Western blot was used to detect protein levels of cytochrome C(Cyt C)in mitochondria and cytoplasm.Activity of Caspase9 was detected by spectrophotometric method,and expression levels of mitochondrial apoptosis related proteins(Bcl-2,Bax and cleaved-Caspase3)were detected by Western blot.Results:PAM can reduced proliferation rate of RL95-2 cells,which was related to drug concentration and duration of action.Compared with blank control group,apoptosis rates of PAM treatment groups were significantly increased(all P<0.05),mitochondrial membrane potential and Cyt C protein levels in mitochondria were significantly decreased(all P<0.05),while Cyt C protein level in cytoplasm was significantly increased(P<0.01).Caspase9 activity was significantly increased(P<0.05),Bcl-2 protein level was significantly decreased(P<0.05),while Bax and cleaved-Caspase3 protein levels were significantly increased(all P<0.01).Conclusion:PAM may promote apoptosis of RL95-2 cells by activating mitochondrial apoptosis pathway.

关 键 词：白术多糖 子宫内膜癌 细胞凋亡 线粒体凋亡途径 

分 类 号：R737.33[医药卫生—肿瘤]