Department of Urology, The Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, 710004, Shaanxi, China.
Department of Obstetrics and Gynecology, Xijing Hospital, Fourth Military Medical University, Xi'an, 710032, Shaanxi, China.
J Bioenerg Biomembr. 2021 Apr;53(2):191-202. doi: 10.1007/s10863-021-09888-2. Epub 2021 Mar 8.
Renal cell carcinoma (RCC) is one of the most common renal malignancies in the urinary system. Numerous studies have demonstrated that miRNAs can regulate tumorigenesis and progression. This study aims to investigate the role and regulatory mechanism of miR-6838-5p in RCC. Our study confirmed that miR-6838-5p was upregulated in human RCC tissues (30/42, 77.43%, P < 0.01) and RCC cell lines (P < 0.05) compared to adjacent non-neoplastic tissues and normal renal epithelial cells. In vitro, overexpression of miR-6838-5p enhanced cell proliferation and invasion in human RCC cell lines (ACHN and 786-O), which were detected by CCK-8, Transwell and Colony formation assays (P < 0.05), and knockdown of miR-6838-5p suppressed cell proliferation and invasion (P < 0.05). Results of Bioinformatics analysis combined with Dual-luciferase reporter gene assay demonstrated that miR-6838-5p could bind to Cyclin D binding myb-like transcription factor 1 (DMTF1). In addition, RT-qPCR and Western blotting confirmed that DMTF1 was downregulated in RCC tissues and cell lines. Meanwhile, it was demonstrated that overexpression of miR-6838-5p inhibited DMTF1 level in ACHN cells. Next, we confirmed that DMTF1 overexpression reversed the inhibitory effects of overexpression of miR-6838-5p on phosphatase and tensin homolog (PTEN), tumor protein 53(p53), murine double minute 2 (MDM2) and alternative reading frame (ARF) protein levels in the ARF-p53 signaling pathway. In conclusion, our research showed that miR-6838-5p enhanced the proliferation and invasion of RCC cells by inhibiting the DMTF1/ARF-p53 axis.
肾细胞癌（RCC）是泌尿系统中最常见的肾恶性肿瘤之一。大量研究表明，miRNAs 可以调节肿瘤的发生和发展。本研究旨在探讨 miR-6838-5p 在 RCC 中的作用和调控机制。我们的研究证实，miR-6838-5p 在人 RCC 组织（30/42，77.43%，P<0.01）和 RCC 细胞系中上调（P<0.05）与相邻非肿瘤组织和正常肾上皮细胞相比。在体外，miR-6838-5p 的过表达增强了人 RCC 细胞系（ACHN 和 786-O）的细胞增殖和侵袭，这通过 CCK-8、Transwell 和集落形成实验检测到（P<0.05），而 miR-6838-5p 的敲低抑制了细胞增殖和侵袭（P<0.05）。生物信息学分析结合双荧光素酶报告基因实验的结果表明，miR-6838-5p 可以与细胞周期蛋白 D 结合 myb 样转录因子 1（DMTF1）结合。此外，RT-qPCR 和 Western blot 证实 DMTF1 在 RCC 组织和细胞系中下调。同时，研究表明，miR-6838-5p 的过表达抑制了 ACHN 细胞中 DMTF1 的水平。接下来，我们证实了 DMTF1 的过表达逆转了 miR-6838-5p 过表达对 ARF-p53 信号通路中磷酸酶和张力蛋白同源物（PTEN）、肿瘤蛋白 53（p53）、鼠双微体 2（MDM2）和交替阅读框（ARF）蛋白水平的抑制作用。总之，我们的研究表明，miR-6838-5p 通过抑制 DMTF1/ARF-p53 轴增强了 RCC 细胞的增殖和侵袭。
