鞘磷脂及神经酰胺对人结肠癌细胞(HT-29)的影响

Influence of sphingomyelin and ceramide on human colon HT-29 cells

目的 研究鞘磷脂及其代谢产物神经酰胺对人结肠癌细胞的影响。方法 采用噻唑蓝 (MTT)显色法、细胞核分裂指数和3H TdR掺入试验方法 ,所设剂量分别为鞘磷脂 5、10、2 0、40 μg ml和神经酰胺 6 2 5、12 5、2 5、5 0 μmol L ,分别以无水乙醇和二甲基亚砜 (DMSO)为溶剂对照。结果 鞘磷脂对HT 2 9细胞的生长无明显作用 ,神经酰胺对HT 2 9细胞的生长有明显抑制作用。在MTT试验中 ,不同浓度神经酰胺对HT 2 9细胞的 7d抑制率分别为 3 9%、85 %、98%和98% ;在核分裂指数试验中 ,随着神经酰胺剂量的增高 ,其核分裂指数明显降低 ;在3H TdR掺入试验中 ,可见随着神经酰胺剂量的增高 ,3H TdR掺入到HT 2 9细胞中明显的减少 ,与阴性对照组相比 ,差异有显著性 (P <0 0 5 )。结论 鞘磷脂对HT 2 9细胞生长无明显作用 ,神经酰胺对HT 2 9细胞的生长增殖有明显抑制作用 ,这可能是鞘磷脂抑制结肠癌的机制之一。

Objective To study the influence of sphingomyelin and ceramide on the growth of human colon HT-29 cells in vitro .Methods Using cell culture tests(MTTtest,index of cellular mitosis,and 3H-TdR incorporation test),we examined the growth of HT-29 cells at four concentrations 5,10,20,40 μg/ml of Sphingomyelin and 6.25、12.5、25、50 μmol/L of ceramide,with a vehicle control(anhydrous alcohol and DMSO v/v).Results Sphingomyelin has shown no effect on the growth of HT-29 cells,however,ceramide can inhibit the growth of HT-29 cells.After treatment with ceramide at various concentrations mentioned above for 7 days,the inhibition rates were 39%,85%,98% and 98% respectively.With the increase of ceramide concentration, 3H-TdR incorporation was decreased in HT-29 cells and the amount of 3H-TdR incorporation was significantly decreased as compared with the vehicle control( P <0.05). Conclusion Ceramide can significantly inhibit the growth of HT-29 cells and it is probably one of the preventive mechanisms of sphingomyelin in colon cancer.