蒽醌修饰物KA-4c触发内质网应激靶向ATF6抗乳腺癌细胞作用机制  

Anthraquinone modifier KA-4c triggers endoplasmic reticulum stress and targets ATF6 against breast cancer cells

作　　者：杨盈盈 许淑妹 赵英丹 陈强健 李俊莹 侯华新[3] 黎丹戎 YANG Ying-ying;XU Shu-mei;ZHAO Ying-dan;CHEN Qiang-jian;LI Jun-ying;HOU Hua-xin;LI Dan-rong(Life Sciences Institute,Guangxi Medical University,Nanning 530021,China;Cancer Hospital,Guangxi Medical University,Nanning 530021,China;College of Pharmacy,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学生命科学研究院,广西南宁530021 [2]广西医科大学肿瘤医学院,广西南宁530021 [3]广西医科大学药学院,广西南宁530021

出　　处：《中国药理学通报》2023年第5期859-867,共9页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82260714);广西自然科学基金重点项目(No 2020GXNSFDA238016);区域性高发肿瘤早期防治研究教育部重点实验室自主课题(No GKE-ZZ202237)。

摘　　要：目的探讨蒽醌修饰物KA-4c抗乳腺癌细胞的作用机制,并采用药物亲和力反应靶标稳定性技术(drug affinity responsive target stability,DARTS)确定其作用靶点。方法MTT法检测细胞活力;HE染色、ER-Tracker Red、电镜研究KA-4c对乳腺癌细胞形态学等的影响;流式细胞术检测KA-4c是否诱导细胞凋亡,Western blot实验检测凋亡蛋白表达;DARTS、细胞热移位分析(cellular thermal shift assay,CETSA)技术确定KA-4c的作用靶点。结果KA-4c对三阴性乳腺癌MDA-MB231细胞的增殖抑制效果最为明显,并可导致内质网和线粒体空泡化而损伤细胞,凋亡率随KA-4c浓度增加而升高,凋亡相关蛋白CHOP、caspase-7随KA-4c浓度增加表达增强。DARTS结果显示,KA-4c可激活内质网蛋白加工信号通路,其中KA-4c与ATF6蛋白结合而耐受蛋白酶水解;CETSA实验结果表明,KA-4c可呈浓度依赖性增强ATF6蛋白的表达。结论KA-4c触发内质网应激诱导乳腺癌细胞凋亡,ATF6可能是KA-4c的作用靶点之一。Aim To explore the mechanism of the effect of anthraquinone modifier KA-4c on breast cancer cells,and determine its action target by drug affinity reaction target stability technique(DARTS).Methods The cell viability was detected by MTT method.The effect of KA-4c on the morphology of breast cancer cells was studied by HE staining,ER-Tracker Red and electron microscope.The apoptosis rate of breast cancer cells induced by KA-4c was detected by flow cytometry.The expression of apoptotic protein was detected by Western blotting.DARTS and CETSA were used to determine the target of KA-4c.Results KA-4c had the most significant inhibitory effect on the proliferation of triple negative breast cancer MDA-MB231 cells,and could cause endoplasmic reticulum and mitochondrial vacuolation to damage the cells.The apoptosis rate and the expression of apoptosis-related proteins CHOP and caspase-7 increased with the increase of KA-4c concentration.DARTS results showed that KA-4c could activate endoplasmic reticulum protein processing signaling pathway,in which KA-4c bound to ATF6 protein and was resistant to protease hydrolysis.The results of CETSA experiments showed that KA-4c could enhance the expression of ATF6 protein in a concentration-dependent manner.Conclusions KA-4 triggers endoplasmic reticulum stress to induce apoptosis in breast cancer cells.ATF6 may be one of the targets of KA-4c.

关 键 词：DARTS 蒽醌修饰物 药物靶点 乳腺癌 内质网应激 ATF6 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R737.9[医药卫生—基础医学] R916.4R979.1