Department of Urology, Tongde Hospital of Zhejiang Province, Hangzhou, China.
Eur Rev Med Pharmacol Sci. 2018 Oct;22(19):6300-6307. doi: 10.26355/eurrev_201810_16040.
To investigate the effect of miR-425 on the proliferation and apoptosis of clear cell renal carcinoma (ccRCA) cells, and to explore the underlying mechanism.
A total of 80 pairs of human clear cell renal carcinoma (ccRCA) and cancer-adjacent normal tissue samples were collected in this study. Human ccRCA cell line (786-O) and normal human kidney cell line (HK-2) were used in cellular research. The expression level of miR-425 was detected in ccRCA tissues and cells, respectively. Target genes of miR-425 were predicted by bioinformatics and verified by luciferase reporter gene assay. Moreover, the role of miR-425 in regulating E2F6 as well as its effect on the proliferation and apoptosis of ccRCA cells were detected.
Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) results showed that the expression of miR-425 was significantly decreased in ccRCA tissues and cells. The proliferation ability and cell cycle of 786-O cells were significantly inhibited after miR-425 overexpression. The percentage of cells in G0/G1 phase was remarkably increased, while the percentage of cells in S and G2/M phases was significantly decreased. Besides, the number of apoptotic cells was significantly increased in the miR-425 intervention group. On-line target gene prediction software indicated that E2F6 was the potential downstream target gene of miR-425. RT-PCR, Western blotting and luciferase reporter gene assay demonstrated that the expression of E2F6 was negatively regulated by miR-425. In addition, subsequent experiments showed that the up-regulation of E2F6 could suppress the inhibitory effect of miR-425 on the proliferation and apoptosis of ccRCA cells.
Our research demonstrated the inhibitory function of miR-425 in ccRCA. Therefore, the miR-425/E2F6 axis was expected to be one of the targets of ccRCA targeted therapy.
研究 miR-425 对肾透明细胞癌（ccRCA）细胞增殖和凋亡的影响，并探讨其作用机制。
本研究共收集 80 对人肾透明细胞癌（ccRCA）组织及其癌旁正常组织标本。采用细胞实验研究，检测 miR-425 在 ccRCA 组织和细胞中的表达水平。采用生物信息学预测 miR-425 的靶基因，并通过荧光素酶报告基因实验进行验证。此外，还检测了 miR-425 对 E2F6 的调控作用及其对 ccRCA 细胞增殖和凋亡的影响。
实时荧光定量聚合酶链反应（qRT-PCR）结果显示，miR-425 在 ccRCA 组织和细胞中的表达明显降低。过表达 miR-425 后，786-O 细胞的增殖能力和细胞周期明显受到抑制，G0/G1 期细胞比例明显增加，S 期和 G2/M 期细胞比例明显减少。此外，miR-425 干预组细胞凋亡数量明显增加。在线靶基因预测软件提示，E2F6 是 miR-425 的潜在下游靶基因。实时荧光定量聚合酶链反应（qRT-PCR）、Western blot 和荧光素酶报告基因实验证实，E2F6 的表达受 miR-425 的负调控。此外，后续实验表明，上调 E2F6 可抑制 miR-425 对 ccRCA 细胞增殖和凋亡的抑制作用。
本研究表明 miR-425 对 ccRCA 具有抑制作用，因此 miR-425/E2F6 轴可能成为 ccRCA 靶向治疗的靶点之一。
