High-throughput sequencing approach for the identification of lncRNA biomarkers in hepatocellular carcinoma and revealing the effect of ZFAS1/miR-150-5p on hepatocellular carcinoma progression

PeerJ

article

Peng Zhu¹  Yongyan Pei²  Jian Yu³  Wenbin Ding¹  Yun Yang¹  Fuchen Liu¹  Lei Liu¹  Jian Huang¹  Shengxian Yuan¹  Zongyan Wang¹  Fangming Gu¹  Zeya Pan¹  Jinzhong Chen⁴  Jinrong Qiu⁵  Huiying Liu⁵ 

[1] Department of Hepatic Surgery ,(III), The Third Affiliated Hospital of Naval Medical University;School of Chemistry and Chemical Engineering, Guangdong Pharmaceutical University;Department of General Surgery, The Third Affiliated Hospital of Naval Medical University;State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University;Department of Biotherapy, The Third Affiliated Hospital of Naval Medical University

关键词: Long non-coding RNA;    Hepatocellular carcinoma;    RNA sequencing;    Progression;    ZFAS1/miR-150-5p;   

DOI  :  10.7717/peerj.14891

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

Aims To screen abnormal lncRNAs and diagnostic biomarkers in the progression of hepatocellular carcinoma through high-throughput sequencing and explore the underlying mechanisms of abnormal lncRNAs in the progression of hepatocellular carcinoma. Methods The transcriptome sequencing was used to analyze the RNA expression profile and identify differentially expressed RNAs. Hub lncRNAs were screened by combining (WGCNA, ceRNA regulatory network, PPI, GO and KEGG analyses, Kaplan-Meier curve analysis, Cox analysis, risk model construction and qPCR). Thereafter, the correlation between the expression of hub lncRNAs and tumor clinicopathological parameters was analyzed, and the hub lncRNAs were analyzed by GSEA. Finally, the effects of hub RNAs on the proliferation, migration and invasion of HepG2 cells were investigated in vitro. Results Compared with the control group, a total of 610 lncRNAs, 2,593 mRNAs and 26 miRNAs were screened in patients with hepatocellular carcinoma. Through miRNA target prediction and WGCNA, a ceRNA was constructed, comprising 324 nodes and 621 edges. Enrichment analysis showed that mRNAs in ceRNA were involved mainly in cancer development progression. Then, the ZFAS1/miR-150-5p interaction pair was screened out by Kaplan Meier curve analysis, Cox analysis and qPCR analysis. Its expression was related to tumor stage, TNM stage and patient age. ROC curve analysis showed that it has a good predictive value for the risk of hepatocellular carcinoma. GSEA showed that ZFAS1 was also enriched in the regulation of immune response, cell differentiation and proliferation. Loss-of-function experiments revealed that ZFAS1 inhibition could remarkably suppress HepG2 cell proliferation, migration and invasion in vitro. Bioinformatic analysis and luciferase reporter assays revealed that ZFAS1 directly interacted with miR-150-5p. Rescue experiments showed that a miR-150-5p inhibitor reversed the cell proliferation, migration and invasion functions of ZFAS1 knockdown in vitro. Conclusion ZFAS1 is associated with the malignant status and prognosis of patients with hepatocellular carcinoma, and the ZFAS1/miR-150-5p axis is involved in hepatocellular carcinoma progression.

【 授权许可】

CC BY