LncRNA LEF1-AS1 silencing diminishes EZH2 expression to delay hepatocellular carcinoma development by impairing CEBPB-interaction with CDCA7.

Abstract

Hepatocellular carcinoma (HCC) is recognized for its high mortality rate worldwide. Based on intensive studies, long non-coding RNA (lncRNA) expression exerts significant effects on tumor suppression. Herein, we investigated the molecular mechanism of lymphoid enhancer-binding factor-1 antisense RNA 1 (LEF1-AS1) in HCC cells. Microarray-based gene expression analysis was adopted to predict and verify the differentially expressed genes in HCC, which predicted cell division cycle-associated 7 (CDCA7) and LEF1-AS1 to be highly expressed in HCC. The expression of LEF1-AS1, CDCA7, CCAAT/enhancer-binding protein beta (CEBPB) and enhancer of zeste homolog 2 (EZH2) was determined by means of reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. LncMap was used to predict the lncRNA-transcription factor-gene interaction in HCC. ChIP, RIP assay and dual luciferase reporter gene assay were employed to verify the relationship between the transcription factor and gene. Silencing of LEF1-AS1 could downregulate CDCA7 expression through CEBPB. Overexpression of LEF1-AS1, EZH2 and CDCA7 promoted proliferation and invasion in HCC cells. LEF1-AS1 promoted CDCA7 expression to further upregulate EZH2. Tumor formation in nude mice was assessed to verify the experimental results. Silencing of LEF1-AS1 inhibited the growth of tumors in vivo. Collectively, silencing LEF1-AS1 inhibited the proliferation and invasion of HCC cells by down-regulating EZH2 through the CEBPB-CDCA7 signaling pathway, which provides scientific evidence for the treatment of HCC.Abbreviations: HCC: Hepatocellular carcinoma; lncRNA: long non-coding RNA; LEF1-AS1: lymphoid enhancer-binding factor-1 antisense RNA 1; EZH2: enhancer of zeste homolog 2; CDCA7: cell division cycle-associated 7; GEO: Gene Expression Omnibus; NC: negative control; oe: overexpressed; RT-qPCR: reverse transcription quantitative polymerase chain reaction; PBS: phosphate buffered saline; HRP: horseradish peroxidase; OD: optical density; RIP: Radioimmunoprecipitation; ChIP: Chromatin immunoprecipitation; WT: wild type.

• Authors: Jun Gao, Chao Dai, Xin Yu, Xiang-Bao Yin, Fan Zhou
• Journal: Cell cycle (Georgetown, Tex.) (Cell Cycle) Vol. 19 Issue 8 Pg. 870-883 (04 2020) ISSN: 1551-4005 [Electronic] United States
• PMID: 32178558 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• CCAAT-Enhancer-Binding Protein-beta
• CDCA7 protein, human
• CEBPB protein, human
• LEF1 protein, human
• Lymphoid Enhancer-Binding Factor 1
• Nuclear Proteins
• RNA, Long Noncoding
• EZH2 protein, human
• Enhancer of Zeste Homolog 2 Protein

Topics

• Adult
• Aged
• Animals
• CCAAT-Enhancer-Binding Protein-beta (genetics, metabolism)
• Carcinogenesis (genetics)
• Carcinoma, Hepatocellular (genetics, metabolism)
• Cell Proliferation (genetics)
• Enhancer of Zeste Homolog 2 Protein (genetics, metabolism)
• Female
• Gene Expression Regulation, Neoplastic
• Gene Silencing
• Hep G2 Cells
• Humans
• Liver Neoplasms (genetics, metabolism)
• Lymphoid Enhancer-Binding Factor 1 (genetics)
• Male
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Middle Aged
• Nuclear Proteins (genetics, metabolism)
• RNA, Long Noncoding (genetics, metabolism)
• Signal Transduction (genetics)
• Transfection
• Tumor Burden (genetics)
• Xenograft Model Antitumor Assays
• Young Adult