乙酰辅酶A羧化酶1通过调控CyclinD1/CDK4影响肾透明细胞癌细胞增殖  

Acetyl-CoA Carboxylase 1(ACC1)Affects the Proliferation of Clear Cell Renal Cell Carcinoma(ccRCC)by Regulating Cyclin D1/CDK4

作　　者：程婧 倪月莉 AGBANA Yannick Luther 云芳 杨晖 赵雷[3] 李小渝 张雪丹 张巧 杨哲[2] 况应敏[3] 朱月春 CHENG Jing;NI Yue-Li;AGBANA Yannick Luther;YUN Fang;YANG Hui;ZHAO Lei;LI Xiao-Yu;ZHANG Xue-Dan;ZHANG Qiao;YANG Zhe;KUANG Ying-Min;ZHU Yue-Chun(Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Kunming Medical University,Kunming 650500,China;Department of Pathology,First Affiliated Hospital of Kunming Medical University,Kunming 650032,China;Department of Organ Transplantation,First Affiliated Hospital of Kunming Medical University,Kunming 650032,China)

机构地区：[1]昆明医科大学基础医学院生物化学与分子生物学系,昆明650500 [2]昆明医科大学第一附属医院病理科,昆明650032 [3]昆明医科大学第一附属医院器官移植科,昆明650032

出　　处：《中国生物化学与分子生物学报》2021年第6期743-751,共9页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.81960462,81760455,31960145);云南省应用研究基金(No.2018FE468(-001))资助。

摘　　要：乙酰辅酶A羧化酶1(acetyl-CoA carboxylase1,ACC1)是脂肪酸合成途径的限速酶。ACC1与多种代谢性疾病和癌症密切相关。然而,ACC1在肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)中的作用及机制却未见报道。本研究以肾透明细胞癌786-O和Caki-1细胞为对象,探讨ACC1异常表达对肾透明细胞癌增殖的影响及其作用机制。首先通过红油-O-染色发现,相比HK2(human kidney-2)细胞,786-O和Caki-1细胞中的脂肪含量明显增加。检索TCGA数据库分析发现,ACC1的蛋白质水平在肾透明细胞癌组织中的表达显著高于正常肾组织(P<0.001)。由分期分级分析发现,临床TNM各期的ACC1蛋白表达均显著高于正常组织。且ACC1表达水平越高,病理分级越高。而ACC1的mRNA水平高表达与肾透明细胞癌病人的不良预后呈正相关。Western印迹结果显示,与对照组HK2细胞相比,在786-O和Caki-1细胞中,ACC1的表达明显增加。通过慢病毒载体构建ACC1敲低的稳转细胞株,由红油-O-染色结果显示,ACC1敲低可显著降低786-O和Caki-1细胞的脂肪含量。CCK-8和平板克隆结果显示,ACC1低表达可显著降低786-O和Caki-1细胞的增殖和克隆形成能力。流式细胞周期结果显示,ACC1敲低可使细胞周期G0/G1期阻滞,并且抑制细胞周期蛋白D1(cyclinD1)和CDK4表达。以上结果表明,ACC1在肾透明细胞癌中异常高表达,并通过上调细胞周期蛋白D1和CDK4表达调控细胞周期进程,促进肿瘤增殖,提示不良预后,ACC1有望成为ccRCC干预治疗的潜在新靶点。Acetyl-CoA carboxylase(ACC)is the rate limiting enzyme of fatty acid synthesis pathway.Studies have shown that ACC1 is implicated in a variety of metabolic diseases and cancer.However,the role and mechanism of action of ACC1 in clear cell renal cell carcinoma(ccRCC)have not been reported.In this study,786-O and Caki-1 clear cell renal carcinoma cells were used as research objects to investigate the effect of abnormal expression of ACC1 on their proliferation and unravel the underlying mechanism.Red oil-O-staining results showed that the lipid content of 786-O and Caki-1 cells was significantly higher than that of human kidney 2(HK2)cells.By searching TCGA database,we found that the expression of ACC1 proteins in ccRCC was significantly higher than that in normal renal tissues(P<0.001).Plus,ACC1 protein expression in all clinical TNM stages was significantly higher than that in normal tissues,and the higher the expression of ACC1,the higher the pathological grade.Furthermore,high expression of ACC1 mRNA is positively correlated with poor prognosis in ccRCC patients.Western blotting analysis showed that the expression of ACC1 in 786-O and Caki-1 cells was significantly higher than that in HK2 cells.The results of red oil-O-staining showed that knocking down ACC1 could significantly reduce the lipid content of 786-O and Caki-1 cells.The results of CCK-8 assays and clonogenicity analysis showed that knocking down ACC1 could significantly reduce the proliferation and colony forming ability of 786-O and Caki-1 cells.Flow cytometry analysis showed that after knocking down ACC1,the cell cycle was blocked at the G0/G1 phase and the expression of Cyclin D1 and CDK4 was inhibited.In conclusion,we found that ACC1 is abnormally overexpressed in ccRCC and it favors proliferation by regulating the expression of Cyclin D1 and CDK4.Therefore,ACC1 is a potential target for the treatment of ccRCC.

关 键 词：乙酰辅酶A羧化酶1 脂代谢 肾透明细胞癌 癌症基因组图谱 增殖 

分 类 号：R34[医药卫生—基础医学]