Inhibitory effect of tumor suppressor p33ING1b and its synergy with p53 gene in hepatocellular carcinoma

doi:10.3748/wjg.v11.i13.1903

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Apr 7, 2005 (publication date) through Nov 19, 2024

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

Liver Cancer

World J Gastroenterol. Apr 7, 2005; 11(13): 1903-1909
Published online Apr 7, 2005. doi: 10.3748/wjg.v11.i13.1903

Inhibitory effect of tumor suppressor p33^ING1b and its synergy with p53 gene in hepatocellular carcinoma

Zhi Zhu, Jing Lin, Jian-Hui Qu, Mark A. Feitelson, Can-Rong Ni, Fang-Mei Li, Ming-Hua Zhu

Zhi Zhu, Jing Lin, Jian-Hui Qu, Can-Rong Ni, Fang-Mei Li, Ming-Hua Zhu, Department of Pathology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China

Mark A. Feitelson, Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA, USA

Author contributions: All authors contributed equally to the work.

Correspondence to: Ming-Hua Zhu, Department of Pathology, Changhai Hospital, Second Military Medical University, 174 Changhai Road, Shanghai 200433, China. mhzhu2000@hotmail.com

Telephone: +86-21-25074604 Fax: +86-21-25074604

Received: July 20, 2004
Revised: July 21, 2004
Accepted: September 4, 2004
Published online: April 7, 2005

Abstract

AIM: To investigate the inhibitory effect of tumor suppressor p33^ING1b and its synergy with p53 gene in hepatocellular carcinoma (HCC).

METHODS: Recombinant sense and antisense p33^ING1b plasmids were transfected into hepatoma cell line HepG2 with lipofectamine. Apoptosis, G₀/G₁ arrest, cell growth rate and cloning efficiency in soft agar of HepG2 were analyzed after transfection. In three hepatoma cell lines with different endogenous p53 gene expressions, the synergistic effect of p33^ING1b with p53 was analyzed by flow cytometry and luciferase assay was performed to detect the activation of p53 downstream gene p21^WAF1/CIP1. In addition, the expression and mutation rates of p33^ING1b in HCC tissues were measured by immunohistochemistry and polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP).

RESULTS: Overexpression of p33^ING1b inhibited cell growth of HepG2, induced more apoptosis and protected cells from growth in soft agar. Combined transfer of p33^ING1b and p53 gene promoted hepatoma cell apoptosis, G₀/G₁ arrest and elevated expression of p21^WAF1/CIP1. Immunostaining results showed co-localized p33^ING1b with P53 protein in HCC tissues and there was a significant relation between protein expression rates of these two genes (P<0.01). Among 28 HCC samples, p33^ING1b presented a low gene mutation rate (7.1%).

CONCLUSION: p33^ING1b collaborates with p53 in cell growth inhibition, cell cycle arrest and apoptosis in HCC. Loss or inactivation of p33^ING1b normal function may be an important mechanism for the development of HCC retaining wild-type p53.

Keywords: Gene p33^ING1b; Gene p53; Apoptosis; Cell cycle arrest; Gene p21^waf1; Liver neoplasm