College of Basic Medicine, Hubei University of Chinese Medicine, Wuhan, China.
China Key Laboratory of TCM Resource and Prescription, Hubei University of Chinese Medicine, Wuhan, China.
J Cell Biochem. 2020 Feb;121(2):1504-1513. doi: 10.1002/jcb.29385. Epub 2019 Sep 9.
MicroRNA-96 (miR-96) is a vertebrate conserved microRNA which plays important roles in various cancers including renal cell carcinoma (RCC). However, its function and mechanism in RCC are still unclear. In this study, miR-96 was found to be downregulated in RCC based on The Cancer Genome Atlas datasets analyses, and its target genes, which predicted by TargetScan, were investigated. Among these target genes, neuronal pentraxin 2 (NPTX2) was upregulated more than 15-fold in RCC, and moreover, closely related to patient survival. To validate its targeting of NPTX2 experimentally, reverse transcription polymerase chain reaction, Western blot analysis, and dual-luciferase assays were performed, and results of these assays demonstrated that miR-96 inhibited expression of NPTX2 through a single 3'-untranslated region targeting site. Furthermore, transfection assays in RenCa and 786-O cells showed miR-96 and small interfering RNA of NPTX2 inhibited cell proliferation, migration, and invasion and overexpression of NPTX2 recovered the inhibition of miR-96. In conclusion, the present study reveals a novel regulatory mechanism of miR-96 on NPTX2 expression in RCC, and the potential of miR-96 as a RCC tumor repressor deserves further investigation.
miR-96(微小 RNA-96)是一种脊椎动物保守的 microRNA,在包括肾细胞癌(RCC)在内的各种癌症中发挥重要作用。然而,其在 RCC 中的功能和机制尚不清楚。本研究基于癌症基因组图谱数据集分析发现 miR-96 在 RCC 中下调,并且研究了其靶基因,这些靶基因是通过 TargetScan 预测的。在这些靶基因中,神经元五肽 2(NPTX2)在 RCC 中上调超过 15 倍,并且与患者的生存密切相关。为了实验验证其对 NPTX2 的靶向作用,进行了逆转录聚合酶链反应、Western blot 分析和双荧光素酶报告基因检测,这些检测结果表明 miR-96 通过单一 3'非翻译区靶位点抑制 NPTX2 的表达。此外,在 RenCa 和 786-O 细胞中转染实验表明,miR-96 和 NPTX2 的小干扰 RNA 抑制细胞增殖、迁移和侵袭,而过表达 NPTX2 恢复了 miR-96 的抑制作用。总之,本研究揭示了 miR-96 在 RCC 中对 NPTX2 表达的新调控机制,miR-96 作为 RCC 肿瘤抑制剂的潜力值得进一步研究。