WT1-AS promotes cell apoptosis in hepatocellular carcinoma through down-regulating of WT1

J Exp Clin Cancer Res. 2015 Oct 13:34:119. doi: 10.1186/s13046-015-0233-7.

Abstract

Background: The antisense of the tumor suppressor gene WT1 (WT1-AS) is a long non-coding RNA. The role of WT1-AS in the development of hepatocellular carcinoma (HCC) has not yet been elucidated.

Methods: Quantitative real-time PCR and western blot analyses were used to measure levels of WT1-AS and its related genes in tumor and corresponding adjacent tumor tissues of HCC patients. The effect on HCC cell proliferation and apoptosis was assessed by EdU incorporation assays and PI-Annexin-V staining, respectively. ShRNA and dual-luciferase assays were used to investigate the regulatory relationship between WT1-AS and WT1 in cell lines.

Results: WT1-AS expression correlated negatively with WT1 expression in HCC tumor tissue. Kaplan-Meier curve analysis revealed that WT1-AS expression is a reliable indicator of HCC prognosis. The downregulation of WT1 expression by WT1-AS promoted cell apoptosis by suppressing the JAK/STAT3 signaling pathway. Bioinformatics analysis showed that WT1-AS downregulates WT1 by binding to the TATA region of the WT1 promotor. WT1-AS was also able to reverse WT1-mediated resistance to Dox based chemotherapy in HCC cells.

Conclusions: WT1-AS downregulates WT1 expression in HCC tumors and promotes apoptosis by binding to the promoter region of WT1. Our findings suggest that WT1-AS may function as a tumor suppressor in HCC by reversing the oncogenic effects of WT1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / genetics*
  • Carcinoma, Hepatocellular / genetics*
  • Carcinoma, Hepatocellular / mortality
  • Cell Line, Tumor
  • Cell Proliferation / genetics
  • DNA, Antisense / biosynthesis
  • DNA, Antisense / genetics*
  • Down-Regulation / genetics
  • Female
  • Gene Expression Regulation, Neoplastic
  • Genes, Wilms Tumor*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Janus Kinases / antagonists & inhibitors
  • Janus Kinases / metabolism
  • Liver Neoplasms / genetics*
  • Liver Neoplasms / mortality
  • Male
  • Middle Aged
  • Prognosis
  • Promoter Regions, Genetic / genetics
  • RNA Interference
  • RNA, Long Noncoding / genetics*
  • RNA, Small Interfering
  • STAT3 Transcription Factor / antagonists & inhibitors
  • STAT3 Transcription Factor / metabolism
  • WT1 Proteins / biosynthesis
  • WT1 Proteins / genetics*

Substances

  • DNA, Antisense
  • RNA, Long Noncoding
  • RNA, Small Interfering
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • WT1 Proteins
  • WT1 protein, human
  • Janus Kinases