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World J Gastroenterol. 2006 Jun 14;12(22):3581-4.

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

Author information

1
Department of Toxicology, Public Health College, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081, Heilongjiang Province, China.

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 micromol/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.

PMID:
16773715
PMCID:
PMC4087574
DOI:
10.3748/wjg.v12.i22.3581
[Indexed for MEDLINE]
Free PMC Article

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