内质网应激-IRE1-ASK1-JNK通路在奥沙利铂诱导乳腺癌细胞凋亡中的作用  

Role of endoplasmic reticulum stress-IRE1-ASK1-JNK pathway in the apoptosis of breast cancer cells induced by oxaliplatin

作　　者：厚玉瑾 田稼[2] 李轩 刘新兰 HOU Yujin;TIAN Jia;LI Xuan;LIU Xinlan(Department of Oncology,General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院肿瘤医院肿瘤内科,银川750004 [2]宁夏医科大学总医院精子库,750004

出　　处：《临床肿瘤学杂志》2021年第5期391-397,共7页Chinese Clinical Oncology

摘　　要：目的探讨内质网应激-IRE1-ASK1-JNK通路在奥沙利铂(L-OHP)介导乳腺癌MDA-MB-231细胞凋亡中的作用。方法体外培养乳腺癌MDA-MB-231细胞,分为正常对照组(Control组)、奥沙利铂组(L-OHP组)、抑制剂组(4-PBA组),流式细胞仪检测各组细胞凋亡率;自噬慢病毒检测各组细胞自噬水平;流式细胞仪检测各组细胞线粒体膜电位(MMP)变化,透射电镜观察各组细胞线粒体结构变化;Western blotting检测各组凋亡、自噬、线粒体结构相关蛋白及内质网应激-IRE1-ASK1-JNK通路相关蛋白的表达变化。结果L-OHP对MB-231细胞毒性呈剂量依赖性,随着L-OHP药物浓度的增加,MB-231细胞凋亡率逐渐增加,差异有统计学意义(P<0.05);计算得出IC 20为13.46μmol/L,IC_(50)为20.98μmol/L。与Control组比较,L-OHP组细胞自噬、凋亡水平增加,自噬相关蛋白LC3Ⅱ/LC3Ⅰ比值增加,p62、Beclin1蛋白表达上调,凋亡相关蛋白Bax、Caspase-3表达上调,Bcl-2蛋白表达下调,差异均有统计学意义(P<0.05);与Control组比较,L-OHP组细胞MMP下降,Mfn2、OPA1蛋白表达下调,DRAP1及Cyt C蛋白表达上调,差异均有统计学意义(P<0.05);透射电镜观察可见L-OHP组细胞线粒体空泡变、固缩,基质电子密度增加,嵴排列变紊乱、扭曲;与Control组比较,L-OHP组GRP78、IRE1、p-ASK1、p-JNK蛋白含量增加,差异有统计学意义(P<0.05);而抑制剂组与L-OHP组比较,上述变化均显著缓解。结论L-OHP通过内质网应激-IRE1-ASK1-JNK通路诱导线粒体损伤,促进乳腺癌细胞凋亡,为进一步研究L-OHP化疗耐药性提供了研究思路。Objective To investigate the role of endoplasmic reticulum stress-IRE1-ASK1-JNK pathway in oxaliplatin(L-OHP)-mediated apoptosis of breast cancer MDA-MB-231 cells.Methods Breast cancer MDA-MB-231 cells in vitro,divided into normal control group(Control group),oxaliplatin group(L-OHP group)and inhibitor group(4-PBA group),and the apoptosis rates were measured by flow cytometry.Autophagy self-phased slow virus was used to detect the level of autophagy.The changes of mitochondrial membrane potential(MMP)were detected by flow cytometry,and the changes of mitochondrial structure were observed by transmission electron microscope.Western blotting was used to detect the related proteins expression of apoptosis,autophagy,mitochondrial structural and endoplasmic reticulum stress-IRE1-ASK1-JNK pathway.Results The cytotoxicity of L-OHP to MDA-MB-231 cells was dose-dependent.With the increase of L-OHP concentration,the apoptosis rate of MDA-MB-231 cells has gradually increased,and the difference has statistical significance(P<0.05),and the IC 20(13.46μmol/L)and IC_(50) (20.98μmol/L)were calculated.Compared with the control group,the autophagy and apoptosis levels increased,autophagy-related protein LC3Ⅱ/LC3Ⅰratio increased,p62 and Beclin1 protein expression increased,apoptosis-related protein Bax and Caspase-3 expression increased,Bcl-2 protein expression decreased in L-OHP group(P<0.05).Compared with the control group,the expression of MMP,Mfn2 and OPA1 protein decreased,and the expression of DRAP1 and cytochrome C(Cyt C)protein increased in L-OHP group(P<0.05).Transmission electron microscopy showed that the mitochondria of L-OHP group were vacuolated and pyknosis,the electron density of matrix increased,and the cristae became disordered and twisted.Compared with the control group,GRP78,IRE1,p-ASK1,p-JNK protein increased in L-OHP group,and the difference was statistically significant(P<0.05).Compared with the L-OHP group,the above changes were significantly alleviated in the inhibitor group.Conclusion L-OHP induces m

关 键 词：乳腺癌 奥沙利铂 内质网应激 信号通路 细胞凋亡 

分 类 号：R737.9[医药卫生—肿瘤]