LncRNA H19/miR-29b-3p/PGRN Axis Promoted Epithelial-Mesenchymal Transition of Colorectal Cancer Cells by Acting on Wnt Signaling

Mol Cells. 2018 May 31;41(5):423-435. doi: 10.14348/molcells.2018.2258. Epub 2018 May 10.

Abstract

This investigation was aimed at working out the combined role of lncRNA H19, miR-29b and Wnt signaling in the development of colorectal cancer (CRC). In the aggregate, 185 CRC tissues and corresponding para-carcinoma tissues were gathered. The human CRC cell lines (i.e. HT29, HCT116, SW480 and SW620) and normal colorectal mucosa cell line (NCM460) were also purchased. Si-H19, si-NC, miR-29b-3p mimics, miR-29b-3p inhibitor, si-PGRN and negative control (NC) were, respectively, transfected into the CRC cells. Lucif-erase reporter plasmids were prepared to evaluate the transduction activity of Wnt/β-catenin signaling pathway, and dual-luciferase reporter gene assay was arranged to confirm the targeted relationship between H19 and miR-29b-3p, as well as between miR-29b-3p and PGRN. Finally, the proliferative and invasive capacities of CRC cells were appraised through transwell, MTT and scratch assays. As a result, over-expressed H19 and down-expressed miR-29b-3p displayed close associations with the CRC patients' poor prognosis (P < 0.05). Besides, transfection with si-H19, miR-29b-3p mimic or si-PGRN were correlated with elevated E-cadherin expression, decreased snail and vimentin expressions, as well as less-motivated cell proliferation and cell metastasis (P < 0.05). Moreover, H19 was verified to directly target miR-29b-3p based on the luciferase reporter gene assay (P < 0.05), and miR-29b-3p also bound to PGRN in a direct manner (P < 0.05). Finally, addition of LiCl (Wnt/β-catenin pathway activator) or XAV93920 (Wnt/β-catenin pathway inhibitor) would cause remarkably altered E-cadherin, c-Myc, vimentin and snail expressions, as well as significantly changed transcriptional activity of β-catenin/Tcf reporter plasmid (P < 0.05). In conclusion, the lncRNA H19/miR-29b-3p/PGRN/Wnt axis counted a great deal for seeking appropriate diagnostic biomarkers and treatment targets for CRC.

Keywords: EMT; PGRN; Wnt signaling; colorectal cancer; lncRNA H19; miR-29b-3p.

Publication types

  • Comparative Study

MeSH terms

  • Antigens, CD
  • Biomarkers
  • Cadherins / biosynthesis
  • Cadherins / genetics
  • Cell Line
  • Cell Line, Tumor
  • Cell Movement
  • Colon / cytology
  • Colorectal Neoplasms / pathology*
  • Epithelial-Mesenchymal Transition*
  • Follow-Up Studies
  • Gene Expression Regulation, Neoplastic
  • Genes, Reporter
  • Humans
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intercellular Signaling Peptides and Proteins / physiology*
  • MicroRNAs / genetics
  • MicroRNAs / physiology*
  • Neoplasm Invasiveness
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / physiology*
  • Prognosis
  • Progranulins
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / physiology*
  • Recombinant Proteins / metabolism
  • Snail Family Transcription Factors / biosynthesis
  • Snail Family Transcription Factors / genetics
  • Transduction, Genetic
  • Vimentin / biosynthesis
  • Vimentin / genetics
  • Wnt Signaling Pathway*

Substances

  • Antigens, CD
  • Biomarkers
  • CDH1 protein, human
  • Cadherins
  • GRN protein, human
  • H19 long non-coding RNA
  • Intercellular Signaling Peptides and Proteins
  • MIRN29 microRNA, human
  • MicroRNAs
  • Neoplasm Proteins
  • Progranulins
  • RNA, Long Noncoding
  • Recombinant Proteins
  • Snail Family Transcription Factors
  • Vimentin