琥珀酸脱氢酶B亚基对卵巢癌细胞线粒体DNA拷贝数影响的研究  

Study on the effect of succinate dehydrogenase B subunit on the mitochondrial DNA copy number in ovarian cancer cells

作　　者：陈立兰[1] 狄文[1,2,3] Chen Lilan;Di Wen(Department of Obstetrics and Gynecology,Renji Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 200127;Shanghai Key Laboratory of Gynecologic Oncology,Shanghai 200127;State Key Laboratory of Oncogenes and Related Genes,Shanghai Cancer Institute,Renji Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 200127)

机构地区：[1]上海交通大学医学院附属仁济医院妇产科,上海200127 [2]上海市妇科肿瘤重点实验室,上海200127 [3]癌基因及相关基因国家重点实验室,上海200127

出　　处：《现代妇产科进展》2022年第3期161-165,共5页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金面上项目(No:81974454,81772770)。

摘　　要：目的:探讨琥珀酸脱氢酶B亚基(SDHB)对卵巢癌线粒体DNA(mtDNA)相对拷贝数的影响。方法:Real-time PCR法检测人卵巢良性肿瘤组织、卵巢癌组织中SDHB mRNA水平,检测mtDNA相对拷贝数(mtND2/HBB)和线粒体转录因子A(TFAM)水平。siRNA干扰SKOV3和A2780中SDHB表达,real-time PCR检测干扰前后mtND2/HBB、TFAM和DNA依赖蛋白激酶催化亚基(DNA-PKcs,也称作PRKDC)水平;CCK-8法检测癌细胞对顺铂的化疗耐药性。Western blot法检测p-p38和DNA损伤标记蛋白γ-H2AX蛋白水平。结果:人卵巢癌组织中SDHB表达低于卵巢良性肿瘤组织,mtND2/HBB和TFAM水平高于卵巢良性肿瘤组织。siRNA干扰SKOV3和A2780中SDHB表达后,mtND2/HBB和PRKDC显著升高,TFAM水平升高不显著。SDHB低表达显著上调p-p38蛋白水平,显著降低γ-H2AX蛋白水平。结论:SDHB在人卵巢癌组织中低表达,SDHB低表达显著增加卵巢癌细胞mtDNA相对拷贝数,提高卵巢癌细胞化疗耐药性,其机制可能与SDHB通过p38 MAPK通路影响DNA损伤修复相关。Objective:To explore the effect of succinate dehydrogenase subunit B(SDHB)on the relative copy number of mitochondrial DNA(mtDNA)in ovarian carcinoma.Methods:The expressions of SDHB mRNA,mtDNA relative copy number(mtND2/HBB)and mitochondrial transcription factor A(TFAM)were measured by real-time PCR in ovarian benign tumor tissues and ovarian carcinoma tissues.SiRNA interference was used to change the level of SDHB in SKOV3 and A2780 ovarian cancer cell lines.Real-time PCR was used to examine the expressions of mtND2/HBB,TFAM and PRKDC.The levels of p-p38 andγ-H2AX were analyzed by Western blot.CCK-8 was used to examine the chemoresistance of cancer cells to cisplatin.Results:SDHB expression in ovarian carcinoma tissues was lower than that in ovarian benign tumor tissues.MtND2/HBB and TFAM were higher than that in ovarian benign tumor tissues.After SDHB was interfered by siRNA in SKOV3 and A2780 cells,mtND2/HBB was increased significantly,TFAM was also increased but not significantly.The low expression of SDHB significantly increased the mRNA level of PRKDC and the protein level of p-p38,decreased the protein level ofγ-H2AX.Conclusion:SDHB expressed lower in ovarian carcinoma tissues,low expression of SDHB significantly increased the relative copy number of mtDNA and improved the chemoresistance in ovarian cancer cells,which may be related to the effect of SDHB on DNA damage repair through P38MAPK pathway.

关 键 词：琥珀酸脱氢酶B亚基 线粒体DNA DNA依赖蛋白激酶催化亚基 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]