柠檬酸合成酶对卵巢癌SKOV3细胞上皮间质转化的影响  

Influence of citrate synthase on epithelial-mesenchymal transformation of ovarian cancer SKOV3 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 Jiao Tong University,Shanghai 200127,China;Shanghai Key Laboratory of Gynecologic Oncology,Shanghai 200127,China;State Key Laboratory of Oncogenes and Related Genes,Shanghai 200127,China)

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：《现代肿瘤医学》2022年第17期3065-3068,共4页Journal of Modern Oncology

基　　金：国家自然科学基金面上项目(编号:81974454);国家重点研发计划(编号:2021YFC2700400)。

摘　　要：目的:探讨柠檬酸合成酶(citrate synthase,CS)对上皮性卵巢癌(epithelial ovarian cancer,EOC)细胞SKOV3上皮间质转化(epithelial-mesenchymal transformation,EMT)及生物学行为的影响。方法:利用Lipofectamine 2000体外瞬时转染化学法合成的CS siRNA,通过实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)和Western blot检测转染效率;Transwell实验检测转染前后SKOV3细胞侵袭和迁移能力的变化;Western blot检测转染前后上皮标记分子E-cadherin、间质标记分子Vimentin蛋白水平和Wnt通路关键分子β-catenin蛋白水平,实时荧光定量PCR检测转染前后EMT相关转录因子Slug、Snail和Twist的mRNA水平。结果:siRNA干扰序列显著降低SKOV3细胞中CS表达;CS低表达显著抑制SKOV3细胞的侵袭和迁移能力,促进上皮标记分子E-cadherin表达,抑制间质标记分子Vimentin表达,降低Wnt通路关键分子β-catenin表达;EMT相关转录因子Snail和Twist表达显著降低(P<0.001,P<0.01),Slug表达下降不显著(P>0.05)。结论:CS低表达显著抑制卵巢癌SKOV3细胞EMT,其机制可能是通过降低Snail和Twist转录因子实现的,为卵巢癌的转移治疗提供初步的实验及理论基础。Objective:To investigate the effect of citrate synthase(CS)on epithelial-mesenchymal transformation(EMT)and biological behavior of epithelial ovarian cancer(EOC)SKOV3 cells.Methods:The transfection efficiency of CS siRNA,synthesized by Lipofectamine 2000 transient transfection in vitro was detected by real-time quantitative PCR and Western blot,and the changes of invasion and migration ability of tumor cells before and after transfection were detected by Transwell assay.Western blot was used to detect the protein levels of epithelial marker molecule E-cadherin andβ-catenin and stromal marker molecule Vimentin.Real-time quantitative PCR was used to detect the mRNA level of EMT-related transcription factors Slug,Snail and Twist before and after transfection.Results:The interference sequence of CS siRNA significantly reduced the level of CS in SKOV3 cells.CS silencing significantly reduced cell invasion and migration ability.In CS silenced cells,E-cadherin level was increased,Vimentin andβ-catenin levels level were decreased.EMT-related transcription factor Snail and Twist mRNA levels were significantly decreased(P<0.001,P<0.01),and Slug level was decreased but not significantly(P>0.05).Conclusion:Low expression of CS significantly inhibited EMT transformation in ovarian cancer SKOV3 cells,which may be achieved by reducing Snail and Twist transcription factors,which provides a preliminary experimental and theoretical basis for metastatic therapy of ovarian cancer.

关 键 词：柠檬酸合成酶 卵巢癌 上皮间质转化 细胞侵袭 细胞迁移 

分 类 号：R737.31[医药卫生—肿瘤]