Abstract
BackgroundThis article focuses on the roles and mechanism of lncRNA CRNDE on the progression of HCC.MethodsWe used qRT-PCR to detect the expression of lncRNA CRNDE in HCC cells, normal cells and clinical tissues. MTT assay, FCM analysis, Transwell migration and invasion assay were used to detect the effects of lncRNA CRNDE on cell viability, apoptosis, migration and invasion of HCC cells. The expression of apoptosis-related proteins Bcl-2, Bax, Cleaved Caspase 3, Cleaved Caspase 9, EMT epithelial marker E-cadherin and mesothelial marker Vimentin were analyzed by Western blot. Online prediction software was used to predict the binding sites between lncRNA CRNDE and miR-539-5p, or miR-539-5p and POU2F1 3’UTR. Dual luciferase reporter assay, qRT-PCR and RNA pulldown were used to detect target-relationship between lncRNA CRNDE and miR-539-5p. Dual luciferase reporter assay, qRT-PCR, Western blot and Immunofluorescence were used to detect target-relationship between miR-539-5p and POU2F1. qRT-PCR was used to detect the expression of miR-539-5p and POU2F1 in clinical tissues. Rescue experiments was used to evaluate the association among lncRNA CRNDE, miR-539-5p and POU2F1. Finally, we used Western blot to detect the effects of lncRNA CRNDE, miR-539-5p and POU2F1 on NF-κB and AKT pathway.ResultslncRNA CRNDE was highly expressed in HCC cells and HCC tissues compared with normal cells and the corresponding adjacent normal tissues. lncRNA CRNDE promoted the cell viability, migration and invasion of HCC cells, while inhibited the apoptosis and promoted the EMT process of HCC cells. lncRNA CRNDE adsorbed miR-539-5p acts as a competitive endogenous RNA to regulate POU2F1 expression indirectly. In HCC clinical tissues, miR-539-5p expression decreased and POU2F1 increased compared with the corresponding adjacent normal tissues. lncRNA CRNDE/miR-539-5p/POU2-F1 participated the NF-κB and AKT pathway in HCC.ConclusionlncRNA CRNDE promotes the expression of POU2F1 by adsorbing miR-539-5p, thus promoting the progression of HCC.
Highlights
This article focuses on the roles and mechanism of Long non-coding RNA (lncRNA) Colorectal Neoplasia Differentially Expressed (CRNDE) on the progression of Hepatocellular carcinoma (HCC)
Results lncRNA CRNDE was upregulated in HCC cells and clinical tissues To indentify the roles of lncRNA CRNDE on the development of HCC, we detected the expression level of lncRNA CRNDE in HCC clinical tissues and cells. qRTPCR showed that compared with the adjacent normal tissues, lncRNA CRNDE was highly expressed in HCC tissues (Fig. 1a)
QRT-PCR showed that compared with the normal hepatocytes cells (HL-7702), lncRNA CRNDE was highly expressed in HCC cells (HepG2, Huh-7 and QGY-7703) (Fig. 1b)
Summary
This article focuses on the roles and mechanism of lncRNA CRNDE on the progression of HCC. Hepatocellular carcinoma (HCC) which has a poor prognosis and high mortality rate, is one of the most malignant cancer worldwide. About 750,000 new cases and 700,000 death cases occur every year in the world [1, 2]. HCC ranks as the third leading cause of cancer-related death worldwide [3]. HCC lacks typical clinical symptom in the early stage and is difficult to diagnose early. Once the typical symptom appears, the tumor is in the advanced stage with a poor prognosis and low 5-year survival rate [4]. A deep understanding of the pathogenesis and molecular mechanism are contribute to the diagnosis and treatment of patients with HCC
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