Participations of TGFBRl and FGF9 in the process of miRNA-140-5p suppressing the proliferation of Wilms’ tumor

Abstract

Objective To explore the functions of miRNA-140-5p in modulating the proliferation of Wilms’ tumor and elucidate the roles of TGFBRl and FGF9. Methods miRNA array assay was conducted for exploring the miRNA expression profile of Wilms’ tumor. Quantitative real-time polymerase chain reaction (PCR) was performed for verifying the expression of miRNA-140-5p in Wilms' tumor tissue and adjacent non-tumor tissues. MTS assay was used for cell proliferation. Bioinformatics was used for predicting target genes for miRNA-140-5p and dual-luciferase reporting system for verifying the target gene. Western blot was performed for detecting the expression of TGFBRl. And the supernatant concentration of FGF9 was determined by enzyme-linked immunosorbent assay (ELISA). Results The relative expression of miRNA-140-5p was 1.69±0.83 and 6.37±3.25 in Wilms' tumor tissue and adjacent non-tumor tissues respectively. The expression of miRNA-140-5p decreased significantly (P<0.05). The proliferation of G401 and SK-NEP-1 cells was inhibited by an over-expression of miRNA-140-5p. When the proliferation of miRNA-con transfected (MOCK) cells was equal to 100%, the relative proliferation rate of G401 with highly miRNA-140-5p expression to MOCK was (87±15)% vs (100±20)% and for SK-NEP-1 was (91±12)% vs (100±9)% (P<0.05). TGFBRl and FGF9 were direct target genes for miRNA-140-5p. Highly expression of miRNA-140-5p could suppress TGFBRl expression in G401 and SK-NEP-1 cells. And the concentrations of FGF9 were (478.66±66.32) vs (535.64±78.53) pg/ml for miRNA-140-5p transfected group versus MOCK group; (486.57±71.01) vs (601.26±77.68) pg/ml for SK-NEP-1 (P<0.05). Conclusions miRNA-140-5p can inhibit the proliferation of Wilms' tumor. And TGFBRl and FGF9 may participate in the process of tumor suppression by miRNA-140-5p. Key words: Wilms’ tumor; Gene expression regulation; Proliferation