Long non-coding RNA LINC00858 exerts a tumor-promoting role in colon cancer via HNF4α and WNK2 regulation

Cell Oncol (Dordr). 2020 Apr;43(2):297-310. doi: 10.1007/s13402-019-00490-8. Epub 2019 Dec 28.

Abstract

Background: Long non-coding RNAs (lncRNAs) are known to be frequently dysregulated in many types of human cancer. As yet, however, their roles in colon carcinogenesis have not been fully elucidated. In the current study, we assessed whether lncRNA LINC00858 may be involved in the progression of colon cancer and, in addition, investigated its downstream targets.

Methods: LINC00858 expression in patient-derived colon cancer tissues and in colon cancer cell lines was determined using RT-qPCR. Also, relationships between LINC00858 expression and various clinicopathological characteristics were analyzed. The subcellular localization of LINC00858 was determined using fluorescence in situ hybridization. Interactions between LINC00858 and its downstream targets were first predicted by bioinformatic analysis and, subsequently, confirmed by RNA pull-down, RNA immunoprecipitation, chromatin immunoprecipitation and dual luciferase reporter assays. After in vitro upregulation of LINC00858 and/or silencing of WNK2 and hepatocyte nuclear factor 4α (HNF4α), the biological behavior of colon cancer cells was assessed using 5-ethynyl-2'-deoxyuridine (EdU) incorporation, Transwell invasion and tube formation assays. In vivo cancer growth was evaluated in nude mice.

Results: We found that LINC00858 was highly expressed in primary colon cancer tissues and colon cancer cell lines, and was mainly located in the nucleus. High LINC00858 expression was found to correlate with a poor differentiation, advanced TNM stages and lymph node metastasis. Exogenous overexpression of LINC00858 promoted cell proliferation, invasion and migration of colon cancer cells, and facilitated angiogenesis and tumor growth. In addition, we found that LINC00858 can bind to and upregulate the nuclear transcription factor HNF4α, leading to WNK2 expression downregulation. This, in turn, resulted in the promotion of colon cancer cell growth.

Conclusions: From our data we conclude that LINC00858 acts as a tumor-promoting lncRNA in colon cancer by upregulating HNF4α and downregulating WNK2. Our results may provide novel targets for the treatment for colon cancer.

Keywords: Angiogenesis; Colon cancer; HNF4α; Invasion; LINC00858; Migration; WNK2.

MeSH terms

  • Adult
  • Aged
  • Animals
  • Carcinogenesis / genetics*
  • Cell Line, Tumor
  • Colonic Neoplasms / genetics*
  • Colonic Neoplasms / pathology
  • Female
  • Gene Expression Regulation, Neoplastic*
  • Gene Knockdown Techniques
  • HCT116 Cells
  • HT29 Cells
  • Hepatocyte Nuclear Factor 4 / genetics*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Male
  • Mice, Nude
  • Middle Aged
  • Protein-Serine-Threonine Kinases / genetics*
  • RNA, Long Noncoding / genetics*
  • Xenograft Model Antitumor Assays / methods

Substances

  • Hepatocyte Nuclear Factor 4
  • RNA, Long Noncoding
  • WNK2 protein, human
  • Protein-Serine-Threonine Kinases