POLG抑制剂阻碍线粒体生物合成抑制三阴性乳腺癌细胞迁移和侵袭
POLG inhibitor suppresses migration and invasion of triple-negative breast cancer cells via blocking mitochondrial biogenesis
摘要
Abstract
Objective To investigate the effects of zalcitabine(ddC),a mitochondrial DNA polymerase γ(POLG)inhibitor,on the migration,invasion,and to preliminarily explore mitochondrial biogenesis of human tri-ple-negative breast cancer MDA-MB-231 cells.Methods The effect of ddC on cell viability was detected using the MTT assay.The migration and invasion abilities of the cells were evaluated using the cell scratch and Transwell in-vasion assays.Cell apoptosis was determined using flow cytometry and a V-FITC/PI cell apoptosis detection kit.The protein expression of POLG,NADH dehydrogenase subunit Ⅰ(NADH1),NADH dehydrogenase subunit Ⅱ(NADH2),ATP synthase subunit 6(ATPase6),cytochrome c oxidase subunit Ⅰ(COX-1)and cytochrome c ox-idase subunit Ⅲ(COX-3)were determined using Western blot.The POLG mRNA level and mtDNA copy number were determined using qPCR.The mitochondrial content and ATP levels were determined using MitoTracker Green fluorescent probe staining and an ATP determination kit.MDA-MB-231 cells were transfected with pcDNA3.1-EG-FP-POLG plasmids to overexpress POLG.The inhibitory effects of ddC on cell migration and invasion were detected in POLG-overexpressed MDA-MB-231 cells.Results POLG expression was higher in MDA-MB-231 cells than in normal mammary epithelial cells(MCF-10A)(P＜0.01).ddC inhibited cell viability in a dose-dependent man-ner.ddC inhibited the migration(P＜0.01)and invasion(P＜0.01)of MDA-MB-231 cells;however,it dis-played no significant inhibitory effects on cell viability in normal mammary epithelial cells(MCF-10A)at the same concentration.ddC downregulated the protein(P＜0.01)and mRNA(P＜0.01)levels of POLG,reduced mtD-NA copy number(P＜0.01)and downregulated mtDNA-coded NADH1,NADH2,ATPase6,COX-1 and COX-3 protein expression(P＜0.01)in MDA-MB-231 cells.Furthermore ddC inhibited mitochondrial content(P＜0.01)and ATP(P＜0.01)levels in MDA-MB-231 cells.POLG overexpression increased the migration(P＜0.05)and invasion(P＜0.05)abilities of MDA-MB-231 cells,while ddC did not significantly inhibit the migra-tion and invasion abilities of MDA-MB-231 cells overexpressing POLG.Conclusion ddC downregulates POLG ex-pression in MDA-MB-231 cells and inhibits mitochondrial biogenesis and ATP levels,thereby inhibiting the migra-tion and invasion of MDA-MB-231 cells.关键词
三阴性乳腺癌/迁移/侵袭/POLG抑制剂/线粒体生物合成/ATP合成Key words
triple-negative breast cancer/migration/invasion/POLG inhibitor/mitochondrial biogenesis/ATP synthesis分类
医药卫生引用本文复制引用
刘行,樊双琴,晏小敏,赵仕杰,王荣,沈祥春,周雪,张玥,陈妍..POLG抑制剂阻碍线粒体生物合成抑制三阴性乳腺癌细胞迁移和侵袭[J].安徽医科大学学报,2024,59(10):1720-1728,9.基金项目
国家自然科学基金项目(编号:82273957) （编号:82273957）
贵州省科技计划项目(编号:黔科合平台人才[2021]5632号、黔科合基础-ZK[2023]重点034) （编号:黔科合平台人才[2021]5632号、黔科合基础-ZK[2023]重点034）
贵州省中医药、民族医药科学技术研究课题(编号:QZYY-2022-025) （编号:QZYY-2022-025）
贵州省卫生健康委科学技术基金项目(编号:gzwkj2021-519) （编号:gzwkj2021-519）
贵州医科大学大学生创新创业训练计划项目(编号:S202210660139) National Natural Science Foundation of China(No.82273957) （编号:S202210660139）
Guizhou Province Science and Technology Plan Project(Nos.Qiankehe Platform Talent[2021]5632,Qiankehe Foundation-ZK[2023]Keynote 034) （Nos.Qiankehe Platform Talent[2021]5632,Qiankehe Foundation-ZK[2023]Keynote 034）
Research Project on Traditional Chinese Medicine and Ethnic Medicine Science and Technology in Guizhou Province(No.QZYY-2022-025) （No.QZYY-2022-025）
Science and Technology Fund Project of Guizhou Provincial Health Commission(No.gzwkj2021-519) （No.gzwkj2021-519）
Guizhou Medical University Student Innovation and Entrepreneurship Training Program(No.S202210660139) （No.S202210660139）
