Apoptosis of human colon carcinoma HT-29 cells induced by ceramide

doi:10.3748/wjg.v12.i22.3581

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Jun 14, 2006 (publication date) through Nov 23, 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

Colorectal Cancer

World J Gastroenterol. Jun 14, 2006; 12(22): 3581-3584
Published online Jun 14, 2006. doi: 10.3748/wjg.v12.i22.3581

Apoptosis of human colon carcinoma HT-29 cells induced by ceramide

Xiao-Feng Zhang, Bai-Xiang Li, Chun-Yan Dong, Rui Ren

Xiao-Feng Zhang, Bai-Xiang Li, Rui Ren, Department of toxicology, Public Health College, Harbin Medical University, Harbin 150081, Heilongjiang Province, China

Chun-Yan Dong, Department of Digestive Medicine, Changhai Hospital, Shanghai 200433, China

Author contributions: All authors contributed equally to the work.

Supported by the National Natural Science Foundation of China, No. 30471447

Correspondence to: Bai-Xiang Li, Public Health College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, Heilongjiang Province, China. libaix@ems.hrbmu.edu.cn

Telephone: +86-451-87502830 Fax: +86-451-87502885

Received: January 13, 2006
Revised: February 8, 2006
Accepted: February 18, 2006
Published online: June 14, 2006

Abstract

AIM: To investigate the effect of exogenous ceramide-induced apoptosis on human colon carcinoma HT-29 cells.

METHODS: Light microscope, transmission electron microscope and fluorescence microscope were used to observe the morphology change of apoptosis in HT-29 cells. Agarose gel electrophoresis was performed to detect the DNA fragment. Mitochondrial function was detected by MTT assay. mRNA expression of Bcl-2 family gene members was determined by reverse transcription polymerase chain reaction (RT-PCR) assay.

RESULTS: After C₂-ceramide treatment, typical characteristics of apoptosis, such as nuclear chromatin breakage, apoptotic body and DNA ladder, could be observed. After exposure to 50 μmol/L C₂-ceramide for 12 and 24 h, cell apoptosis was 64.1% and 81.3% respectively, which had a time-and dose-effect relationship. Mitochondrial function started to decrease from 6 h after exposure to ceramide. Meanwhile, ceramide up-regulated or down-regulated the mRNA expression of Bcl-2 family gene members.

CONCLUSION: Ceramide induces apoptosis of human colon carcinoma HT-29 cells by affecting the expression of Bcl-2 family gene members and impacting the mitochondrial function.

Keywords: Ceramide; Apoptosis; Human colon carcinoma cells; Bcl-2 family gene member; Mitochondrial function