Decreased miR-200a-3p is a key regulator of renal carcinoma growth and migration by directly targeting CBL
Decreased miR-200a-3p is a key regulator of renal carcinoma growth and migration by directly targeting CBL
复制标题
miR-200a-3p 的减少是直接靶向 CBL 的肾癌生长和迁移的关键调节因子。
DOI：
10.1002/jcb.27326
复制
发表时间：
2018-12-01
影响因子：
4
通讯作者：
Wang, Cheng
中科院分区：
文献类型：
--
作者：
Ding, Meng;Sun, Xinlei;Wang, Cheng
虽然新的证据表明microRNAs（miRNAs）的失调有助于癌的发生，但其在肾细胞癌（RCC）中的作用机制尚不清楚。本研究旨在分析miR-200 a-3 p表达与肾细胞癌的相关性，了解miR-200 a-3 p在肾细胞癌中潜在的新靶基因、功能及作用机制。首先通过定量实时聚合酶链反应和原位杂交在成对的RCC组织样本中测量MiR-200 a-3 p表达水平。接下来，使用计算机辅助算法、荧光素酶报告基因测定和Western印迹分析的组合来分析潜在的miR-200 a-3 p靶基因。最后，通过5-乙炔基-20-脱氧尿苷、细胞凋亡实验和transwell实验研究了miR-200 a-3 p在体外RCC肿瘤发生中的生物学作用，并利用异种移植小鼠模型研究了miR-200 a-3 p在体内RCC肿瘤发生中的生物学作用。我们的研究结果表明，miR-200 a-3 p在RCC组织中的表达明显低于正常组织，CBL是miR-200 a-3 p的直接靶点。在RCC组织样品中观察到miR-200 a-3 p和CBL之间的负相关性。机制研究表明，在RCC细胞系中，miR-200 a-3 p的异位表达通过直接抑制CBL在体外和体内抑制细胞增殖和迁移，并加强细胞凋亡，而沉默miR-200 a-3 p导致相反的效果。此外，过表达CBL消除了miR-200 a-3 p过表达诱导的效应。综上所述，我们的研究结果表明，miR-200 a-3 p/CBL调节轴是RCC发病机制的一种新机制，并可能作为RCC的候选生物标志物和治疗靶点。
Although emerging evidence has revealed that microRNAs (miRNAs) dysregulation contribute to carcinogenesis, the mechanism underlying their roles in renal cell carcinoma (RCC) is unclear. The purpose of the current study was to analyze the association of miR-200a-3p expression with RCC and to understand potential novel target genes, functions and mechanisms of miR-200a-3p in RCC. MiR-200a-3p expression levels were first measured by quantitative real-time polymerase chain reaction and in situ hybridization in pairs of RCC tissue samples. Next, the potential miR-200a-3p target gene was analyzed using a combination of computer-aided algorithms, luciferase reporter assays and Western blot analysis. Finally, the biological roles of miR-200a-3p in RCC tumorigenesis were investigated both in vitro by 5-ethynyl-20-deoxyuridine, apoptosis assay and transwell assay, as well as in vivo using a xenograft mouse model. Our results demonstrated that miR-200a-3p was remarkably downregulated in RCC tissues compared with normal adjacent tissue, and CBL is a direct target of miR-200a-3p. An inverse correlation between miR-200a-3p and CBL was observed in RCC tissue samples. Mechanistic investigations revealed that ectopic expression of miR-200a-3p in RCC cell lines suppressed cell proliferation and migration and enforced cell apoptosis by directly inhibiting CBL in vitro and in vivo, whereas silencing miR-200a-3p resulted in the opposite effects. Additionally, overexpressing CBL abolished the effects induced by miR-200a-3p overexpression. Taken together, our results show that the miR-200a-3p/CBL regulation axis is a novel mechanism underlying RCC pathogenesis and may serve as a candidate biomarker and therapeutic target in RCC.