摘要
目的探讨外源性神经酰胺诱导人结肠癌HT-29细胞的凋亡作用及对Bcl-2家族蛋白基因表达的影响。方法不同浓度C2-神经酰胺处理HT-29细胞,采用Hoechest荧光染色和琼脂糖凝胶电泳方法检测细胞凋亡,RT-PCR方法检测Bcl-2、Bcl-xl、Bax、Bad和Bid基因表达水平。结果C2-神经酰胺诱导HT-29细胞发生核染色质断裂、出现凋亡小体等典型凋亡改变,50μmol/L神经酰胺作用12和24小时凋亡细胞率分别达到64·7%和81·3%。同时神经酰胺使Bax、Bad和Bid基因表达水平增高,使Bcl-2和Bcl-xl基因表达水平减弱,Bcl-2与Bax比值下降,呈现剂量-效应关系。结论外源性神经酰胺能通过上调或下调Bcl-2家族基因表达水平诱导人结肠癌HT-29细胞发生凋亡。
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 apoptosls-related genes was determined by reverse transcription polymerase chain reaction （RT-PCR） analysis, Results Under the contribution of C2-ceramide, typical apoptosls appearance, such as nuclear chromatin breakage, apoptotic body etc and DNA ladder were observed. Exposing to 50μmol/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 inceased the expression of Bax, Bad and Bid mRNA, and decreased the expression of Bcl-2 and Bcl-xl mRNA. Conclusion Ceramide may induce apoptosis of human colon cancer HT-29 cells via up-regulating or down-regulating the expression of apoptosis-related genes,
出处
《卫生研究》
CAS
CSCD
北大核心
2006年第5期537-540,共4页
Journal of Hygiene Research
基金
国家自然科学基金资助项目(No.30471447)
