[Effect of ceramide on apoptosis of human colon cancer HT-29 cells].
OBJECTIVE
To investigate the effect of apoptosis and mRNA expression of apoptosis-related genes of human colon cancer HT-29 cells induced by exogenous ceramide.


METHODS
After HT-29 cells exposed to C2-ceramide for different time, transmission electron microscope and fluorescence microscope were used to observe the morphology change of cells. DNA agarose gel electrophoresis was applied to detect DNA fragmentation of HT-29 cells. The mRNA expression of apoptosis-related genes was determined by reverse transcription polymerase chain reaction (RT-PCR) analysis.


RESULTS
Under the contribution of C2-ceramide, typical apoptosis appearance, such as nuclear chromatin breakage, apoptotic body etc and DNA ladder were observed. Exposing to 50 micromol/L C2-ceramide for 12h and 24h, the rate of apoptotic cells were 64.7% and 81.3% respectively, which appeared time-dose-relation. Meanwhile, ceramide increased the expression of Bax, Bad and Bid mRNA, and decreased the expression of Bcl-2 and Bcl-xl mRNA.


CONCLUSIONS
Ceramide may induce apoptosis of human colon cancer HT-29 cells via up-regulating or down-regulating the expression of apoptosis-related genes.
Zhang
Xiao-Feng
XF
Department of Toxicology, Public Health College of Harbin Medical University, Harbin 150086, China.
Li
Bai-Xiang
BX
Ren
Rui
R
D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000378:metabolism / Q000473:pathology*; D015536:Down-Regulation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D015854:Up-Regulation
D004791:Enzyme Inhibitors; C064769:N-acetylsphingosine; D019253:Proto-Oncogene Proteins c-bcl-2; D013110:Sphingosine