Nie Fang, Liu Tianming, Zhong Liang, Yang Xianggui, Liu Yunhong, Xia Hongwei, Liu Xiaoqiang, Wang Xiaoyan, Liu Zhicheng, Zhou Li, Mao Zhaomin, Zhou Qin, Chen Tingmei
Department of Laboratory Medicine, Key Laboratory of Diagnostic Medicine, Ministry of Education, Chongqing Medical University, Chongqing 400016, P.R. China.
Department of Medical Oncology and Laboratory of Signal Transduction and Molecular Targeted Therapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Mol Med Rep. 2016 Jan;13(1):83-90. doi: 10.3892/mmr.2015.4555. Epub 2015 Nov 11.
Increasing evidence revealed that miRNAs, the vital regulators of gene expression, are involved in various cellular processes, including cell growth, differentiation, apoptosis and progression. In addition, miRNAs act as oncogenes and/or tumor suppressors. The present study aimed to verify the potential roles of miR148b in human renal cancer cells. miR‑148b was found to be downregulated in human renal cancel tissues and human renal cancer cell lines. Functional studies demonstrated that plasmid‑mediated overexpression of miR‑148b promoted cell proliferation, increased the S‑phase population of the cell cycle and enhanced apoptosis in the 786‑O and OS‑RC‑2 renal cancer cell lines, while it did not appear to affect the total number of viable cells according to a Cell Counting Kit‑8 assay. Subsequently, a luciferase reporter assay verified that miR148b directly targeted mitogen‑activated protein kinase (MAPK) kinase kinase 9 (MAP3K9), an upstream activator of MAPK kinase/c‑Jun N‑terminal kinase (JNK) signaling, suppressing the protein but not the mRNA levels. Furthermore, western blot analysis indicated that overexpression of miR148b in renal cancer cells inhibited MAPK/JNK signaling by decreasing the expression of phosphorylated (p)JNK. In addition, overexpression of MAP3K9 and pJNK was detected in clinical renal cell carcinoma specimens compared with that in their normal adjacent tissues. The present study therefore suggested that miR‑148b exerts an oncogenic function by enhancing the proliferation and apoptosis of renal cancer cells by inhibiting the MAPK/JNK pathway.
越来越多的证据表明,微小RNA(miRNAs)作为基因表达的重要调节因子,参与了包括细胞生长、分化、凋亡和进展在内的各种细胞过程。此外,miRNAs可作为癌基因和/或肿瘤抑制因子。本研究旨在验证miR148b在人肾癌细胞中的潜在作用。研究发现,miR-148b在人肾肿瘤组织和人肾癌细胞系中表达下调。功能研究表明,质粒介导的miR-148b过表达促进了786-O和OS-RC-2肾癌细胞系的细胞增殖,增加了细胞周期中S期细胞的比例,并增强了细胞凋亡,而根据细胞计数试剂盒-8检测,其似乎不影响活细胞总数。随后,荧光素酶报告基因检测证实miR148b直接靶向丝裂原活化蛋白激酶(MAPK)激酶激酶9(MAP3K9),MAP3K9是MAPK激酶/c-Jun氨基末端激酶(JNK)信号通路的上游激活因子,可抑制其蛋白水平而非mRNA水平。此外,蛋白质印迹分析表明,肾癌细胞中miR148b的过表达通过降低磷酸化(p)JNK的表达来抑制MAPK/JNK信号通路。此外,与正常相邻组织相比,临床肾细胞癌标本中检测到MAP3K9和pJNK的过表达。因此,本研究表明,miR-148b通过抑制MAPK/JNK途径增强肾癌细胞的增殖和凋亡,发挥致癌功能。