摘要
目的:探讨NDUFA4介导线粒体活化水平对卵巢癌细胞SKOV3增殖、迁移及侵袭的影响。方法:以慢病毒技术建立SKOV3细胞的NDUFA4过表达和沉默模型;CCK-8检测细胞增殖,细胞划痕检测细胞迁移,Transwell检测细胞侵袭,流式细胞术检测线粒体膜电位和凋亡;Real-time PCR检测线粒体DNA(mtDNA)拷贝数,MT-ND1、MT-ND5、MT-CYB、SDHA、PGC-1α、NRF1和mTFA mRNA的表达;电镜观察线粒体形态,分光光度法检测Complex IV活性水平;Western blot检测PGC-1α、NRF1、mTFA蛋白的表达。结果:SKOV3细胞中实现NDUFA4的过表达和沉默;NDUFA4的过表达提高细胞增殖、侵袭和迁移能力(P<0.05),上调mtDNA拷贝数,上调MT-ND1、MT-ND5、MT-CYB、SDHA、PGC-1α、NRF1和mTFA mRNA水平(P<0.05),上调PGC-1α、NRF1和mTFA蛋白水平(P<0.05),上调线粒体膜电位水平以及Complex IV活性水平(P<0.05),线粒体变多形态正常。而NDUFA4的沉默则相反,线粒体出现空泡化。结论:NDUFA4可能通过加强线粒体的活化和氧化磷酸化的水平在SKOV3细胞中发挥促癌功能。
Objective:To investigate the effect of mitochondrial activation level mediated by NDUFA4 on proliferation,migration and invasion of ovarian cancer SKOV3 cells.Methods:NDUFA4 overexpression and silencing models of SKOV3 cells were established by lentivirus technique.Cell proliferation was detected by CCK-8,cell migration was detected by scratch,cell invasion was detected by Transwell,mitochondrial membrane potential and apoptosis were detected by flow cytometry.The mitochondrial DNA copy number,MT-ND1,MT-ND5,MT-CYB,SDHA,PGC-1α,NRF1 and mTFA mRNA expressions were detected by Real-time PCR.The morphology of mitochondria was observed by electron microscope.The activity level of Complex IV was determined by spectrophotometry.The expressions of PGC-1α,NRF1 and mTFA were detected by Western blot.Results:NDUFA4 was overexpressed and silenced in SKOV3 cells.Overexpression of NDUFA4 increased cell proliferation,invasion and migration(P<0.05),up-regulated mtDNA copy number,and up-regulated mRNA levels of MT-ND1,MT-ND5,MT-CYB,SDHA,PGC-1α,NRF1 and mTFA(P<0.05).The protein levels of PGC-1α,NRF1 and mTFA were up-regulated(P<0.05),and the mitochondrial membrane potential and Complex IV activity levels were up-regulated(P<0.05),mitochondria are pluralistic and normal.The silencing of NDUFA4 was opposite,mitochondria vacuolated.Conclusion:NDUFA4 may play a role in promoting cancer in SKOV3 cells by enhancing mitochondrial activation and oxidative phosphorylation.
作者
孟焕然
董辉
冯帆帆
马鹏珠
田雪敏
石璐璐
杨彩虹
MENG Huanran;DONG Hui;FENG Fanfan;MA Pengzhu;TIAN Xuemin;SHI Lulu;YANG Caihong(Obstetrics and Gynecology,General Hospital of Ningxia Medical University,Cardio-cerebrovascular Hospital,Ningxia Yinchuan 750004,China;General Hospital of Ningxia Medical University,Ningxia Yinchuan 750004,China;Ningxia Medical University,Ningxia Yinchuan 750004,China.)
出处
《现代肿瘤医学》
CAS
2024年第6期1024-1032,共9页
Journal of Modern Oncology
基金
宁夏自然科学基金项目(编号:2021AAC03352)。
