ΔNp63上调3-羟-3-甲戊二酸单酰辅酶A还原酶表达促进食管鳞状细胞癌细胞增殖及迁移  

ΔNp63 promotes proliferation and migration of esophageal squamous cell carcinoma cells by up-regulating the expression of 3-hydroxy-3-methylglutaryl-coenzyme A reductase

作　　者：王卫星[1] 袁扬[2] 刘敬禹[1] 江旭[1] 杨朝爱[1] WANG Wei-xing;YUAN Yang;LIU Jing-yu;JIANG Xu;YANG Chao-ai(Department of Interventional Therapy,Changhai Hospital,Naval Medical University(Second Military Medical University),Shanghai 200433,China;Department of Cardiovascular Surgery,Changhai Hospital,Naval Medical University(Second Military Medical University),Shanghai 200433,China)

机构地区：[1]海军军医大学(第二军医大学)长海医院介入治疗科,上海200433 [2]海军军医大学(第二军医大学)长海医院心血管外科,上海200433

出　　处：《第二军医大学学报》2021年第12期1349-1354,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金面上项目(81572357).

摘　　要：目的探讨N端转录活性域缺失的肿瘤蛋白p63(ΔNp63)调控食管鳞状细胞癌细胞增殖及迁移的分子机制。方法以人食管鳞状细胞癌细胞ECA109为模型,通过慢病毒介导的转基因在细胞中过表达ΔNp63。通过定量PCR及蛋白质印迹法检测3-羟-3-甲戊二酸单酰辅酶A还原酶(HMGCR)表达水平,通过染色质免疫沉淀(ChIP)和萤光素酶报告实验检测ΔNp63蛋白与HMGCR基因5'-非翻译区的结合;在细胞中过表达ΔNp63同时利用RNA干扰抑制HMGCR表达后,通过CCK-8检测细胞增殖情况,流式细胞术检测细胞凋亡情况,Transwell实验检测细胞迁移情况。结果ECA109细胞中过表达ΔNp63后,HMGCR的mRNA和蛋白质表达水平均提高(P均<0.01)。ChIP及萤光素酶报告实验结果提示,ΔNp63识别位点位于HMGCR启动子-728~-747 bp区域。过表达ΔNp63同时抑制HMGCR表达组24、48、72 h时细胞增殖能力均低于单纯过表达ΔNp63组(P均<0.05),细胞凋亡水平高于单纯过表达ΔNp63组[(26.9±1.9)%vs(16.6±1.5)%,P<0.01],细胞的迁移率低于单纯过表达ΔNp63组[(33.4±3.1)%vs(52.2±2.6)%,P<0.01]。结论ΔNp63通过上调HMGCR转录表达促进食管鳞状细胞癌细胞的增殖及迁移。Objective To investigate the molecular mechanism of tumor protein 63 with truncated transactivation domain(ΔNp63)in regulating the proliferation and migration of esophageal squamous cell carcinoma(ESCC)cells.Methods With ECA109 cells as a model,ΔNp63 was overexpressed in cells by transgene mediated with lentivirus.The expression level of 3-hydroxy-3-methylglutaryl-coenzyme A reductase(HMGCR)was detected by quantitative polymerase chain reaction and Western blotting,and the binding ofΔNp63 and the 5'-untranslated region(5'-UTR)of HMGCR was detected by chromatin immunoprecipitation(ChIP)and luciferase activity reporter assay.ΔNp63 was overexpressed in cells and the expression of HMGCR was inhibited by RNA interference.Cell proliferation was detected by cell counting kit 8(CCK-8),apoptosis was detected by flow cytometry,and cell migration was detected by Transwell chamber culture.Results The mRNA and protein expression levels of HMGCR were increased after overexpressingΔNp63 in ECA109 cells(both P<0.01).The results of the ChIP and luciferase activity reporter assay showed that theΔNp63 recognition site was located in the-728 to-747 bp region of the HMGCR promoter.The cell proliferation ability of the group overexpressingΔNp63 and inhibiting HMGCR expression was significantly lower than that of the group overexpressingΔNp63 alone at 24,48 and 72 h(all P<0.05);the level of apoptosis was(26.9±1.9)%,which was significantly higher than that of the group overexpressingΔNP63 alone([16.6±1.5]%,P<0.01);meanwhile,the cell migration rate was significantly lower than that of the group overexpressingΔNp63 alone[(33.4±3.1]%vs[52.2±2.6]%,P<0.01).ConclusionΔNp63 can promote the proliferation and migration of ESCC cells by up-regulating the transcriptional expression of HMGCR.

关 键 词：食管肿瘤 鳞状细胞癌 ΔNP63 甲羟戊酸途径 3-羟-3-甲戊二酸单酰辅酶A还原酶 

分 类 号：R735.1[医药卫生—肿瘤]