MicroRNA-140-5p通过蛋白激酶C亚型&epsilon;促进肾癌细胞增殖

刘伟红 余肖 任雨 刘开泰 林晨 王萍*

doi:10.16098/j.issn.0529-1356.2019.02.008

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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

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*

作者信息 +

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*

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文章历史 +

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。

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.

导出引用

刘伟红余肖任雨刘开泰林晨王萍. MicroRNA-140-5p通过蛋白激酶C亚型ε促进肾癌细胞增殖[J]. 解剖学报. 2019, 50(2): 185-191 https://doi.org/10.16098/j.issn.0529-1356.2019.02.008

LIU Wei-hong YU Xiao REN Yu LIU Kai-tai LIN Chen WANG Ping. MicroRNA-140-5p promotes cell proliferation via protein kinase C isoform ε in renal cell carcinoma [J]. Acta Anatomica Sinica. 2019, 50(2): 185-191 https://doi.org/10.16098/j.issn.0529-1356.2019.02.008

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