解剖学报

解剖学报 ›› 2019, Vol. 50 ›› Issue (2): 185-191.doi: 10.16098/j.issn.0529-1356.2019.02.008

• 肿瘤生物学 • 上一篇    下一篇

MicroRNA-140-5p通过蛋白激酶C亚型ε促进肾癌细胞增殖

刘伟红1 余肖1 任雨2,4 刘开泰3 林晨1 王萍1,4*   

  1. 1.宁波大学医学院 人体解剖学与组织学胚胎学科系，浙江 宁波 315211; 2.宁波市泌尿肾病医院肾癌实验室，浙江 宁波 315000;   3.宁波大学医学院李惠利医院宁波市医疗中心，浙江 宁波 315041; 4. 宁波大学泌尿肾病研究所肾癌实验室，浙江 宁波 315000
  • 收稿日期:2018-05-15 修回日期:2018-07-25 出版日期:2019-04-06 发布日期:2019-04-06
  • 通讯作者: 王萍 E-mail:89117968@qq.com
  • 基金资助:
    国家自然科学基金;浙江省科学卫生计划;宁波自然科学基金

MicroRNA-140-5p promotes cell proliferation via protein kinase C isoform ε in renal cell carcinoma 

LIU Wei-hong1 YU Xiao1 REN Yu 2,4 LIU Kai-tai3 LIN Chen1 WANG Ping 1,4*   

  1. 1.Department of Human Anatomy, Histology and Embryology, Medical School, Ningbo University, Zhejiang Ningbo 315211, China;  2.Laboratory Kidney Carcinoma, Ningbo Urology and Nephrology Hospital, Zhejiang Ningbo 315000, China;  3.Ningbo City Medical Treatment Center Lihuili Hospital, Medical School, Ningbo University,  Zhejiang Ningbo 315041, China;  4. Laboratory of Kidney Carcinoma, Urology and Nephrology Institute of Ningbo University, Zhejiang Ningbo 315000, China
  • Received:2018-05-15 Revised:2018-07-25 Online:2019-04-06 Published:2019-04-06
  • Contact: WANG Ping E-mail:89117968@qq.com

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摘要：

目的 探讨microRNA-140-5p(miR-140-5p)在肾癌发生中的作用。 方法 Real-time PCR方法检测肾癌细胞（Caki-1、ACHN、OS-RC-2和786-O）中miR-140-5p的表达。MTS，流式细胞术和Western blotting检测细胞增殖，细胞周期和相关蛋白的改变。 结果 降低Caki-1细胞中miR-140-5p的水平后，细胞增殖被抑制，G0/G1期细胞数量显着增加，cyclin D、cyclin E和CDK2、CDK4也随之减少；此外，增加786-O细胞中miR-140-5p的水平后，促进体外细胞增殖和体内肿瘤发生。基因芯片结果发现，蛋白激酶C亚型ε（PKCε)基因在改变Caki-1和786-O的细胞中显著性升高。siPKCε在降低肾癌细胞（RCC)中PKCε蛋白表达的同时，还能显著性反转抑制物/模拟物诱导的细胞增殖和细胞周期的改变。 结论 miR-140-5p通过PKCε促进肾癌细胞增殖。

关键词: MicroRNA-140-5p, 肾癌细胞, 细胞增殖, 细胞周期, 实时定量聚合酶链反应, 人

Abstract:

Objective To explore the role of micro RNA-140-5p(miR-140-5p) in renal cell carcinoma (RCC) tumorigenesis. Methods The expression of miR-140-5p was determined by Real-time PCR in RCC (Caki-1, ACHN, OS-RC-2 and 786-O), and compared with kidney cortex proximal tubule cell (HK-2). After Caki-1 and 786-O cells were transfected with miR-140-5p inhibitor or mimic, the cell proliferation, cell cycle and related proteins were evaluated by MTS assay, flow cytometry and Western blotting. Results Caki-1 and 786-O showed the highest and lowest expression of miR-140-5p respectively. After the level of miR-140-5p in Caki-1 cells was decreased, cell proliferation was inhibited and cell number in G0/G1 phase was increased significantly, along with the reduction of cyclin D, E and CDK2, 4. Furthermore, the increasing level of miR-140-5p in 786-O cells promoted cell proliferation in vitro and tumorigenesis in vivo.Moreover, microarray demonstrated that protein kinase C isoform ε(PKCε), a markedly enhanced gene in RCC cells with the changed level of miR-140-5p. The combination treatment with down-regulation of PKCε, inhibitor/mimic induced changes of cell proliferation and cell cycle arrests were marked reversed in Caki-1 and 786-O cells.Conclusion These result indicate that miR-140-5p may promote cell proliferation via PKCεin RCC.

Key words: Micro RNA-140-5p, Renal cell carcinoma, Cell proliferation, Cell cycle, Real-time PCR, Human