结直肠癌中miR-330-3p-RUVBL1信号轴对肿瘤细胞增殖、凋亡的影响  

Effect of miR-330-3p-RUVBL1 Signal Axis on Tumor Proliferation and Apoptosis in Colorectal Cancer

作　　者：黄开禹 穆磊 王向阳[1] 王辉[1] Huang Kaiyu;Mu Lei;Wang Xiangyang(Department of Gastrointestinal Surgery,The Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430014,China;Department of Gastrointestinal Surgery,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属武汉市中心医院胃肠外科,武汉430014 [2]华中科技大学同济医学院附属同济医院胃肠外科,武汉430030

出　　处：《华中科技大学学报（医学版）》2020年第5期517-523,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省自然科学基金资助项目(No.2019CFC845)。

摘　　要：目的探讨结直肠癌(colorectal cancer,CRC)中miR-330-3p-RUVBL1信号轴对肿瘤增殖、凋亡的影响。方法分析Oncomine数据库中结直肠癌RUVBL1 mRNA的表达情况;利用Kaplan-Meier生存曲线分析RUVBL1 mRNA表达水平与CRC患者生存预后之间的相关性;免疫组化法检测临床CRC标本癌组织和正常结直肠组织RUVBL1蛋白的表达情况。荧光素酶报告基因实验分析RUVBL1与miR-330-3p的靶向关系;荧光定量聚合酶链式反应(qRT-PCR)检测CRC临床标本癌组织和正常结直肠组织miR-330-3p的表达情况;利用脂质体转染结直肠癌HT-29细胞,过表达或沉默miR-330-3p的表达;运用CCK-8法检测过表达miR-330-3p对HT-29细胞活力的影响和对顺铂敏感性变化,利用流式细胞仪检测过表达miR-330-3p对HT-29凋亡的影响;利用Western blot检测转染miR-330-3p后HT-29细胞中RUVBL1蛋白的表达情况和细胞凋亡相关蛋白的表达情况。结果数据库分析结果显示RUVBL1 mRNA在CRC中高表达;RUVBL1基因的表达水平与结直肠癌患者生存预后之间存在相关性,RUVBL1高表达患者的总体生存率明显低于低表达RUVBL1的患者(P=0.0077);免疫组化结果显示RUVBL1在CRC组织中的表达水平较正常结直肠组织明显增高;qRT-PCR结果显示CRC临床标本癌组织中miR-330-3p的表达水平明显低于正常结直肠组织(P<0.01);过表达miR-330-3p的HT-29细胞中RUVBL1的表达明显下调,细胞内促凋亡蛋白Bax、cleaved-Caspase-3的蛋白表达水平均上调,而Bcl-2表达水平下降;过表达miR-330-3p的HT-29细胞凋亡增加,且对顺铂的敏感性增强。结论RUVBL1在CRC中高表达,其表达水平与CRC患者生存预后相关;RUVBL1是miR-330-3p的潜在靶点,miR-330-3p可能通过抑制RUVBL1表达促进结直肠癌细胞凋亡。Objective To investigate the effect of miR-330-3 p-RUVBL1 signal axis on tumor proliferation and apoptosis in colorectal cancer(CRC).Methods The expression of RUVBL1 mRNA in colorectal cancer was analyzed in Oncomine online database.Kaplan-Meier survival curve was used to analyze the correlation between RUVBL1 mRNA expression and survival,prognosis of colorectal cancer patients.Immunohistochemistry was conducted to detect the expression of RUVBL1 protein in cancer tissues and normal colorectal tissues.RT-PCR was performed to detect the expression of miR-330-3 p in CRC tissues.The targeting relationship between RUVBL1 and miR-330-3 p was analyzed with luciferase reporter gene.The effect of miR-330-3 p on tumor proliferation and apoptosis was observed after miR-330-3 p liposome was transfected into colorectal cancer HT-29 cells.Results The expression level of RUVBL1 in CRC tissue was significantly increased,and there was a correlation between the expression level of RUVBL1 gene and the survival,prognosis of patients with colorectal cancer.The overall survival rate of patients with high expression of RUVBL1 was lower than that in patients with low expression RUVBL1(P=0.0077).Immunohistochemical results showed that the expression of RUVBL1 in CRC tissues was significantly higher than that in normal colorectal tissues.RT-PCR results showed that the expression level of miR-330-3 p in CRC tissues was significantly lower than that in normal colon tissues(P<0.01).Correlation analysis showed that there was a significant correlation between miR-330-3 p level and RUVBL1 protein expression level,and miR-330-3 p transfection could inhibit the proliferation of human colorectal cancer HT-29 cells and promote cell apoptosis.The protein expression levels of pro-apoptotic proteins Bax and cleaved-Caspase-3 were up-regulated,while the expression level of Bcl-2 was down-regulated.Conclusion RUVBL1 is highly expressed in colorectal cancer tissues,and miR-330-3 p is a potential target of RUVBL1.Down-regulation of miR-330-3 p expression

关 键 词：结直肠癌 Oncomine数据库 miR-330-3p RUVBL1 

分 类 号：R735.3[医药卫生—肿瘤]