MiR ‐30a‐5p Regulates the Proliferation of Wilms Tumor Cells Through CyclinE2

OBJECTIVES: To explore the role and mechanism of miR-30a-5p in the progression of Wilms tumor. METHODS: TARGET-WT and GSE50505 matrix files were downloaded to analyze the differences of miRs between Wilms tumor and normal renal tissues. miRDB, miRTarBase, and TARGET-WT databases were applied to predict the target genes of miR. qRT-PCR was applied to determine the expression of the target miR in Wilms tumor/adjacent normal renal tissues as well as G401/WiT49/HEK-293T cells. The interaction of miR-30a-5p with cyclinE2 (CCNE2) was confirmed by correlation analysis, qRT-PCR, Western blot, and double luciferase assay respectively. CCK-8, colony formation, cell cycle assay, Transwell, and wound healing assay were applied to determine the influence of miR-30a-5p on the proliferation or migration of G401/WiT49 cells. Furthermore, a mouse transplanted tumor model was constructed to explore the function of miR-30a-5p on Wilms' tumor growth in vivo. RESULTS: miR-30a-5p was down-regulated and CCNE2 was up-regulated in Wilms tumor tissues and G401/WiT49 cells compared to the control, respectively. Double luciferase reporter assay showed that miR-30a-5p could target the 3'UTR of CCNE2. Over-expression or knockdown of miR-30a-5p modulated the CCNE2 mRNA and protein expressions as well as the expressions of p-Rb and E2F1 downstream of CCNE2 in G401/WiT49 cells. The rescue experiment indicated that miR-30a-5p regulates the proliferation of G401/WiT49 through CCNE2. Furthermore, upregulation of miR-30a-5p restrained the growth rate of transplanted tumors in nude mice. CONCLUSIONS: miR-30a-5p participates in regulating the proliferation of Wilms tumor cells by suppressing the CCNE2/p-Rb/E2F1 axis.