MiRNA-1180 suppresses HCC cell activities via TRAF1/NF-κB signaling pathway

Feng Zheng,Zheng Wang

doi:10.1590/fst.26219

Abstract

Abstract The miRNA-1180 anti-cancer effects in hepatocellular carcinoma cell were studied. 33 hepatocellular carcinoma patients were collected. In cancer and adjacent normal tissue from patients, TRAF1 protein and miR-1180 expression was determined by HE staining, IHC and RT-PCR methods. In the cell experiments, HepG2 cells were divided into three groups: NC group, BL group and miRNA group. The cells of NC group were un-treated; BL group were transfected with empty camer; miRNA group were transfixed with miRNA-1180. We determinates cell proliferation rate of difference groups, measuring the cell apoptosis rate and cell cycle of difference groups by flow cytometry, detected cell invasion and migration abilities of difference groups by transwell and wound healing assay and evaluating relative proteins expressions by WB assay. Compared with Normal liver tissue, cell infiltrations were significantly increased. miR-1180 was negative correlation with TRAF. The cell proliferation rate of miRNA group was significantly lower than that of NC group; The cell apoptosis and G1 phase rates were significantly difference among three groups; however, P53 and P21 protein expressions were significantly increased in miRNA groups. Over-expression miRNA-1180 had effect to inhibit HepG2 cell proliferation, invasion, migration and improve HepG2 cell apoptosis by regulation TRAF1/NF-κB signaling pathway.

Highlights

Hepatocellular carcinoma (HCC) a kind of cancer that were higher death rate
HepG2 was purchased from ATCC, DMEM and Fetal Bovine Serum were purchased from HyClone; miRNA-1180 transfection group (miRNA)-1180 (Kingsy, China); LipofectamineTM RNAiMAX (Invitrogen, USA); Opti-MEM (Gibco, USA); Cell proliferation assay kit (Promega, USA); Cell apoptosis and cycle assay kit (KeyGEN, USA); Tumor necrosis factor receptor associated factor 1 (TRAF1), NF-κb, P53, P21, MMP-2 and MMP-9 anti-body (Abcam, USA)
HCC is high degree of malignancy and poor prognosis, even with treatment of surgery combined with chemotherapy, the treatment effect is still poor, postoperative HCC high recurrence rate and metastasis is a key factor affecting the treatment effect and prognosis, there is an urgent need for new and effective therapeutic targets. miRNA has a wide range of gene regulatory functions, can regulate various aspects of gene activity, participate in a series of biological processes such as embryonic development, cell proliferation, apoptosis and energy metabolism (Esquela-Kerscher & Slack, 2006)

Summary

Introduction

Hepatocellular carcinoma (HCC) a kind of cancer that were higher death rate. The HCC treatments were significant progressing, the 5 year sruvival rate remained low (Jemal et al, 2011), this was relatived with higher proliferation of HCC. MicroRNAs (miRNAs) were a novel class of RNA, conserved non coding sigle stranded small molecules weidely, in recent years, the miRNA was contained 22 nucleotide, the main effect of miRNA is targeted to mRNA, and degradating or inhibiting targeted mRNA translation, suppressing the mRNA expression and regulating targeted mRNA expression and somatic development, apoptosis, proliferation and differentiation (Volk & Shomron, 2011; Jebbawi et al, 2014; Huang et al, 2014; Chiang et al, 2015). We wanted to prove the effects of miRNA-1180 to HCC and explain the mechanism

Methods

Results

Discussion

Conclusion