MicroRNA-301a-3p Suppressed The Progression Of Hepatocellular Carcinoma Via Targeting VGLL4

Objective:Dysregulation of micro RNAs(miRNAs)is involved in a variety of biological process including tumorigenesis.MiR-301a-3p has been reported to be an onco-miRNA in various types of cancer,like breast cancer,malignant melanoma,and pancreatic cancer.However,the role of miR-301a-3p in hepatocellular carcinoma(HCC)remains largely incomplete.The current study is to investigate the biological function and clinical significance of miR-301a-3p in HCC.Method:1.Quantitative Real-time PCR(qRT-PCR)was used to detect the mRNA expression of miR-301a-3p in a panel of HCC cell lines and in 36 pairs of fresh HCC tissue and adjacent noncancerous tissues,and analysed the relationship between the expression of miR-301a-3p and clinical pathological features in HCC.2.The miR-301a-3p mimics/inhibitors and non-target scrambles were transfected into Hep G2 and SMMC-7721 cells using lipofectamine 2000,when cell confluence reached 50–60%,according to the manufacturer’s instructions.3.The CCK-8 assay was conducted to evaluate the impact of miR-301a-3p on cell proliferation.4.The colony formation assay was conducted to evaluate the impact of miR-301a-3p on cell sensitivity to oxaliplatin.5.Transwell assay was conducted to detect the ability of invasion in HCC cells after modulating miR-301a-3p expression level.6.Dual-luciferase reporter assay was conducted to detect the target of miR-301a-3p,western blotting and site-directed mutagenesis assays were performed to further verify it.7.A series of rescue experiments were conducted to testify the involvement of VGLL4 in the function of miR-301a-3p in promoting HCC progression.Results:1.The expression of miR-301a-3p significantly associated with lymph node metastasis status in HCC.2.MiR-301a-3p was upregulated in HCC tissues and cell lines,and higher miR-301a-3p expression predicted poor prognosis in HCC patients.3.MiR-301a-3p overexpression enhanced the ability of proliferation,invasion,and chemoresistance in HCC cell lines.4.VGLL4 is a direct miR-301a-3p target.5.MiR-301a-3p upregulates the transcription activity of TEAD via modulating VGLL4.6.VGLL4 is involved in the biological function of miR-301a-3p in promoting the progression of HCC.Conclusion:The results of the present study indicated that miR-301a-3p displayed an oncogenic function during HCC development,via direct binding to the 3’-UTR of VGLL4,thereby enhanced the transcription activity of TEAD.Our findings suggested that the miR-301a-3p-VGLL4 signaling axis might be a potential therapeutic target for HCC..