Preliminary Study On The Effect Of MiR-145-5p On Hepatoblastoma By Regulating The PI3K/AKT Signaling Pathway

Objective: To investigate the expression difference of miR-145-5p between hepatoblastoma and paracancer tissues,and to determine the effect of miR-145-5p on hepatoblastoma and its possible molecular mechanism.Methods: A total of 10 pairs of tissues and paracancarcinoma tissues that were surgically collected in our hospital after pathological examination.The differential expression of miR-145-5p between hepatoblastoma and paracancer tissues was screened out based on GEO database.The expression levels of miR-145-5p in tissues(hepatoblastoma tissue,paracarcinoma tissue)and cells(normal liver cell line Lo-2,hepatoblastoma cell line HepG2 and hepatoma cell line Huh-7)were determined by q PCR.The miR-145-5p analogs was transfected into HepG2 cell line withrelatively low expression of miR-145-5p for overexpression.The proliferation and apoptosis of transfected cells were detected by CCK8 and flow cytometry,respectively.The potential binding target genes of miR-145-5p and their cellular function and enrichment of signaling pathways were predicted by bioinformatics analysis..Western blot assay was used to detect the expression changes of key molecules and potential target genes in the PI3K/AKT signaling pathway of hepatoblastoma after miR-145-5p overexpression.Results: 1.The expression of miR-145-5p in hepatoblastoma tissues was significantly down-regulated compared with that in paracancer samples(P < 0.05);2.The expression level of miR-145-5p in HepG2 cell lines was significantly down-regulated compared with that in Lo-2 and Huh-7 cell lines(P < 0.05),which was consistent with the scenario of miR-145-5p expression trendency in collected clinical samples.3.After miR-145-5p mimic was transfecting into HepG2 cells,the proliferation of HepG2 cells was decreased and apoptosis was increased compared with control cells(P < 0.05).4.After miR-145-5p overexpression,the protein expressions of key members such as p-PI3 K and p-AKT in PI3K/AKT signaling pathway)were decreased(P < 0.05),and the m RNA expressions of its potential target genes(RPS6KB1,NRAS,ANGPT2,IRS1,and ITGB8)were decreased(P < 0.05).Conclusion: The up-regulation of miR-145-5p in hepatoblastoma may promote the apoptosis of hepatoblastoma cells and inhibit the occurrence and development of hepatoblastoma by inhibiting the activity of PI3K/AKT signaling pathway.