Restored expression of the tumor suppressor gene RUNX3 reduces cancer stem cells in hepatocellular carcinoma by suppressing Jagged1-Notch signaling

Shin Ichi Nishina, Hidenori Shiraha, Yutaka Nakanishi, Shigetomi Tanaka, Minoru Matsubara, Nobuyuki Takaoka, Masayuki Uemura, Shigeru Horiguchi, Junro Kataoka, Masaya Iwamuro, Takahito Yagi, Kazuhide Yamamoto

Research output: Contribution to journalArticle

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Abstract

Runt-related transcription factor 3 (RUNX3) is a candidate tumor suppressor gene that is downregulated in various cancers. In the present study, we analyzed the regulatory function of RUNX3 on Jagged-1 (JAGl) expression and cancer stem cell (CSC) signaling in hepatocellular carcinoma (HCC). Eleven HCC cell lines and 30 human HCC tissues were used. RUNX3 and JAG1 expression levels were analyzed by immunoblotting and immunohistochemistry. Ectopic RUNX3 expression was induced by introducing RUNX3 cDNA into the RUNX3-negative HCC cell line Hep3B and Huh7 cells. Furthermore endogenous RUNX3 expression was knocked down by RUNX3 siRNA in SK-Hep-1 cells. In order to analyze JAGl transcriptional regulation, we conducted reporter assays, chromatin immunoprecipitation (ChIP) assays and electrophoretic mobility shift assays (EMSAs). Tumorigenicity was analyzed using a SCID mouse liver injection model. An inverse correlation was observed between RUNX3 expression and JAG1 expression in most HCC cell lines and tissues. Restoring RUNX3 expression decreased the expression of JAG1 in Hep3B and Huh7 cells, whereas JAG1 expression was upregulated in RUNX3 siRNA-treated SK-Hep-1 cells. Reporter assays, ChIP assays and EMSAs revealed that RUNX3 directly bound to the transcriptional regulatory region of JAG1 and suppressed JAG1 transcription. Moreover, RUNX3 restoration downregulated CSCs by suppressing JAG1-mediated Notch signaling. The tumorigenic capacity of RUNX3-expressing Hep3B cells was lower compared to that of control Hep3B cells. RUNX3 expression suppressed JAG1 expression and resulted in downregulation of tumorigenesis by suppression of JAG1-mediated CSCs.

Original languageEnglish
Pages (from-to)523-531
Number of pages9
JournalOncology Reports
Volume26
Issue number3
DOIs
Publication statusPublished - Sep 2011

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Neoplastic Stem Cells
Tumor Suppressor Genes
Hepatocellular Carcinoma
Down-Regulation
Chromatin Immunoprecipitation
Electrophoretic Mobility Shift Assay
Cell Line
Small Interfering RNA
Transcription Factor 3
SCID Mice
Nucleic Acid Regulatory Sequences
Immunoblotting
Carcinogenesis
Complementary DNA
Immunohistochemistry
Injections
Liver
Neoplasms

Keywords

  • Cancer stem cell
  • Hepatocellular carcinoma
  • Jagged-1
  • Notch signaling
  • Runt-related transcription factor 3

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Restored expression of the tumor suppressor gene RUNX3 reduces cancer stem cells in hepatocellular carcinoma by suppressing Jagged1-Notch signaling. / Nishina, Shin Ichi; Shiraha, Hidenori; Nakanishi, Yutaka; Tanaka, Shigetomi; Matsubara, Minoru; Takaoka, Nobuyuki; Uemura, Masayuki; Horiguchi, Shigeru; Kataoka, Junro; Iwamuro, Masaya; Yagi, Takahito; Yamamoto, Kazuhide.

In: Oncology Reports, Vol. 26, No. 3, 09.2011, p. 523-531.

Research output: Contribution to journalArticle

Nishina, Shin Ichi ; Shiraha, Hidenori ; Nakanishi, Yutaka ; Tanaka, Shigetomi ; Matsubara, Minoru ; Takaoka, Nobuyuki ; Uemura, Masayuki ; Horiguchi, Shigeru ; Kataoka, Junro ; Iwamuro, Masaya ; Yagi, Takahito ; Yamamoto, Kazuhide. / Restored expression of the tumor suppressor gene RUNX3 reduces cancer stem cells in hepatocellular carcinoma by suppressing Jagged1-Notch signaling. In: Oncology Reports. 2011 ; Vol. 26, No. 3. pp. 523-531.
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