成髓细胞瘤转录因子第1亚型参与调控肾透明细胞癌增殖及其机制

Involvement of myeloblastosis proto-oncogene like 1 in regulation of proliferation of renal clear cell carcinoma and mechanism

目的探讨肾透明细胞癌组织中成髓细胞瘤转录因子第1亚型(MYBL1)的变化及微小RNA(miRNA,miR)-509-5p的作用机制。方法收集2018年1月至2019年6月在广西医科大学第一附属医院泌尿外科确诊的肾透明细胞癌及癌旁组织40例及配对的癌旁组织40例。将肾组织分为miR-509-5p mimics组(miR组)和miR-NC组(对照组),实时定量反转录聚合酶链反应(RT-qPCR)检测肾癌及癌旁组织中MYBL1及miR-509-5p的变化,转染miR-509-5p mimics构建miR-509-5p过表达的786-O细胞后,采用细胞计数试剂盒(CCK-8)检测细胞的增殖能力、蛋白质印迹法(Western blot)检测转染后的细胞中MYBL1蛋白表达量变化。组间比较采用t检验。结果与癌旁组织比较,肾癌组织中MYBL1的mRNA(-1.501±0.311,t=4.825,P<0.01)表达上调,而miR-509-5p(4.421±0.515,t=8.583,P<0.01)表达下调;双荧光素酶基因报告表明MYBL1为miR-509-5p的直接靶基因(0.564±0.017,t=13.355,P<0.01),差异均有统计学意义。结论肾透明细胞癌的临床进展可能与miR-509-5p和MYBL1表达量相关,且过表达miR-509-5p可相应的敲低MYBL1,进而减弱肾透明细胞癌的增殖能力。因此miR-509-5p可能通过对MYBL1调控参与肾透明细胞癌增殖。

Objective To investigate the changes of myeloblastosis proto-oncogene like 1(MYBL1)and the action mechanism of microRNA(miRNA,miR)-509-5p.Methods The clinical data of 40 cases of renal clear cell carcinoma and adjacent tissues diagnosed in the Department of Urology,the First Affiliated Hospital of Guangxi Medical university from January 2018 to June 2019 were collected,and the expression levels of MYBL1 and mir-509-5p were detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR).The miR-509-5p was overexpressed in 786-0 cells via transfection with miR-509-5p mimics.Cell counting kit-8(CCK-8)was used to detect the proliferation of the cells,and Western blotting was used to determine the level of MYBL1 protein in the transfected cells.Student t test was used for data processing.Results As compared with the adjacent tissues,MYBL1 mRNA expression(-1.501±0.311,t=4.825,P<0.01)was up-regulated in renal cancer tissues,and miR-509-5p(4.421±0.515,t=8.583,P<0.01)was down-regulated.The dual-luciferase reporter assay showed that MYBL1 was the direct target of miR-509-5p(0.564±0.017,t=13.355,P<0.01).Conclusion The clinical progression of renal clear cell carcinoma may be related to the expression levels of mir-509-5p and MYBL1,and the overexpression of miR-509-5p can correspondingly knock down MYBL1,thereby reducing the proliferation capacity of renal clear cell carcinoma.Therefore,miR-509-5p may be involved in the proliferation of renal clear cell carcinoma by regulating MYBL1.