三氧化二砷对三阴乳腺癌干细胞MDA-MB-231细胞株凋亡和自噬作用机制研究

Mechanism of Arsenic Trioxide on Apoptosis and Autophagy in Triple-negative Breast Cancer Stem Cell MDA-MB-231 Cell Line

目的:探讨三氧化二砷对TNBC干细胞MDA-MB-231细胞株凋亡和自噬作用机制。方法:取第2代培养5天的干细胞进行实验,共分为4组:对照组、三氧化二砷作用组、Rapa联合三氧化二砷组和3-MA联合三氧化二砷组。观察检测细胞凋亡率、细胞自噬、蛋白表达情况。结果:三氧化二砷作用48 h,细胞总凋亡率为40%,采用3MA抑制细胞自噬活性,三氧化二砷诱导的细胞凋亡率降低约50%,Rapa可促使三氧化二砷诱导细胞凋亡增高约50%。与对照组相比,三氧化二砷组、Rapa联合三氧化二砷组、3-MA联合三氧化二砷组胞内荧光强度均较高。3-MA联合三氧化二砷组Bax、Beclin-1、P62蛋白表达高于三氧化二砷组、Rapa联合三氧化二砷组,Bcl-2蛋白表达低于三氧化二砷组、Rapa联合三氧化二砷组(P<0.05)。结论:三氧化二砷诱导的MDA-MB-231干细胞中自噬和细胞调亡现象共存且可能受共同基因的调控,三氧化二砷诱发自噬可促进细胞调亡。

Objective:To investigate the mechanism of arsenic trioxide on apoptosis and autophagy in triple negative breast cancer(TNBC)stem cell MDA-MB-231 cell line.Methods:The stem cells cultured in the second generation were taken for 5 and divided into four groups:control group,arsenic trioxide acting group,Rapa combined with arsenic trioxide group and 3-MA combined with arsenic trioxide group.The apoptosis rate,autophagy and protein expression were observed.Results:For 48 h,the total apoptosis was 40%.When 3MA inhibited the autophagic activity,the apoptosis induced by arsenic trioxide decreased by about 50%,and Rapa promoted the arsenic trioxide induced apoptosis increased by about 50%.Compared with the control group,the intracellular fluorescence intensity was higher in arsenic trioxide,Rapa combined with arsenic trioxide,and 3-MA combined with arsenic trioxide.Bax,Beclin-1 and P62 protein expression were higher in 3-MA and arsenic trioxide group and Rapa group,and Bcl-2 protein expression was lower than the group and Rapa with arsenic trioxide group(P<0.05).Conclusion:Arsenic trioxide induced autophagy and cell apoptosis coexist in MDA-MB-231 stem cells and may be regulated by common genes.Arsenic trioxide induced autophagy can promote cell apoptosis.