MiR-15a-5p在肾癌中的表达分析及临床意义研究  

Expression and clinical significance of miR-15a-5p in renal cell carcinoma

作　　者：金露[1,2,3] 李一帆[1,2,3] 何韬[2,3] 胡佳[2,3] 刘家驹[2,3] 石敏[2] 亓英[2] 桂耀庭[2] 毛向明[2,3] 杨尚琪[2,3] 来永庆[2,3] 

机构地区：[1]安徽医科大学,合肥230032 [2]北京大学深圳医院泌尿外科,广东深圳518036 [3]广东省男性生殖与遗传重点实验室,广东深圳518036

出　　处：《中华临床医师杂志（电子版）》2016年第8期4-9,共6页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家自然科学基金(81101922);深圳市科技研发资金知识创新计划基础研究项目(JCYJ20130402114702124;JCYJ20150403091443304;JCYJ20150403091443329);广东省重点医学专科经费

摘　　要：目的检测肾癌及癌旁配对组织、肾癌及正常肾细胞系中mi R-15a-5p的表达水平,比较配对组织及细胞系之间的表达水平,分析表达水平与患者临床特征之间的关系,探究mi R-15a-5p作为肾癌标记物应用于临床的可能性。方法提取肾癌及正常肾细胞系、36对肾癌与配对癌旁组织标本中的总RNA,经反转录、q PCR检测mi R-15a-5p的表达,使用统计软件分析组织间、细胞系之间mi R-15a-5p的表达差异,分析mi R-15a-5p表达水平与患者临床特征间的关系。通过转染mi R-15a-5p干扰敲低肾癌细胞中mi R-15a-5p表达水平,利用流式细胞术评估其对肾癌细胞的凋亡影响。利用SPSS 19.0进行配对t检验及Fisher确切概率检验。结果 mi R-15a-5p在肾癌组织中表达水平明显高于癌旁正常组织(P<0.05),癌组织中表达水平为癌旁组织的(9.95±0.92)倍。肾癌细胞系(786-O、ACHN)中mi R-15a-5p表达量分别是正常肾细胞系(HEK-293T)的(8.40±1.74)倍、(2.94±0.11)倍(P<0.05)。标本中共有26例(72.2%)肾癌标本mi R-15a-5p表达上调,癌组织中mi R-15a-5p表达水平与肾癌临床分期相关(P<0.05)。敲低mi R-15a-5p表达水平可诱导肾癌细胞凋亡,786-O细胞凋亡率为(14.01±0.81)%(敲低mi R-15a-5p)和(2.47±0.28)%(阴性对照)(P<0.05),ACHN细胞凋亡率为(20.30±0.47)%(敲低mi R-15a-5p)和(11.45±0.61)%(阴性对照)(P<0.05)。结论 mi R-15a-5p在肾癌组织及细胞系中表达明显上调,且和肾癌分期相关,mi R-15a-5p可调节肾癌细胞凋亡,提示其在肾癌的发生发展中起着一定的作用,有作为肾癌标记物应用于临床的可能。Objective To detect the expression level of mi R-15a-5p in paired tissues and cell lines. Then compared the expression of mi R-15a-5p in paired tissues and different cell lines, and analyzed the relationship between the expression level of mi R-15a-5p and clinical characteristics to explore the possibility to be used a biomarker for mi R-15a-5pin renal cell carcinoma. Methods Total RNA was isolated from RCC cell lines and 36 paired tissues, then RT-PCR and q PCR were performed to detect the expression of mi R-15a-5p. Statistic analysis was performed to explore the differences of mi R-15a-5p＇s expression level and the relationships between the expression of mi R-15a-5p and clinical characteristics of patients. The expression level of mi R-15a-5p in RCC was knocked down by transfection of mi R-15a-5p inhibitor. And whether mi R-15a-5p could affect the apoptotic rate of RCC cells was assessed by performing flow cytometry assay. Results Mir-15a-5p was up-regulated in RCC tissues compared with adjacent normal tissues（P〈0.05）. The expression level of mi R-15a-5p in RCC tissues was 9.95±0.92 times of adjacent normal tissues. And the expression level of mi R-15a-5p in RCC cell lines（786-O and ACHN） was 8.40±1.74 and 2.94±0.11 times（P〈0.05） of HEK-293 T cell line, respectively. In 26 paired tissues（72.2%）, the expression level of mi R-15a-5p in RCC tissues was higher than adjacent normal tissues. And the expression level of mi R-15a-5p was associated with the clinical stage of RCC（P〈0.05）. The loss of mi R-15a-5p could induce RCC cell apoptosis： the apoptosis rates of 786-O cells were（14.01±0.81）% vs.（2.47±0.28）% in cells with low expression of mi R-15a-5p vs. negative control and（20.30±0.47）% vs.（11.45±0.61）% in ACHN cells with low expression of mi R-15a-5p vs. negative control. Conclusion Mi R-15a-5p is up-regulated in RCC tissues and cell lines, and the level is associated with clinical stage of RCC. Mi R-15a-5p could regulate the apoptosis of RCC cells, which indica

关 键 词：癌 肾细胞 肿瘤分期 微小RNA miR-15a-5p 

分 类 号：R737.11[医药卫生—肿瘤]