C2－神经酰胺对人间皮瘤细胞凋亡的诱导作用

摘要

Objective In this research ,the effects of exogenous C 2-ceramide on the induction of apop-tosis of mesothelioma cells in vitro and related important proteins are investigated .Mtehods Mesothelioma cells were treated with various doses of C 2-ceramide for different duration .Cell viability were analyzed by cell count-ing kit-8 assay.Morphological changes of apoptosis of mesothelioma cells were observed by Diff Quik staining . Apoptosis of mesothelioma cells was also detected by Caspase -3 assay and lactate dehydrogenase(LDH)assay. Related important proteins involved in the signal transduction of apoptosis were detected by western blot .Results In vitro,C2-ceramide demonstrated a dose -and time-dependent inhibition of cell proliferation .Apoptotic bodies were observed by Diff Quik staining .The antiproliferative effect of 80 μm C2-ceramide was paralleled with an increase in Caspase -3 activity.LDH assay showed that C2-ceramide at a concentration of 80μm signifi-cantly promoted cells death .After treated by C2-ceramide,the expressions of Bax and the phosphorylated JNK in mesothelioma cells were increased , however , the expression of phosphorylated ERK 1/2 kinase was decreased . Conclusion Our results indicate that C 2-ceramide induced apoptosis of malignant mesothelioma cells in vitro . This anti-tumor affect is achieved by adjusting related important proteins .%目的：研究C2－神经酰胺对体外培养的人间皮瘤细胞的凋亡及其相关分子的影响。方法采用CCK－8法检测不同剂量的C2－神经酰胺作用于间皮瘤后不同时间的细胞增殖情况；采用Diff Quik染色观察细胞形态学上的凋亡改变。用Fluorescent Assay 法检测间皮瘤细胞的Caspase－3活性；通过Western blot检测细胞凋亡发生过程中的关键蛋白的表达。结果 C2－神经酰胺对间皮瘤细胞的抑制呈现剂量依从性和时间依从性。 Diff Quik 染色观察到凋亡小体。 C2－神经酰胺能增加间皮瘤细胞Caspase－3的活性。 C2－神经酰胺可使间皮瘤细胞Bax蛋白、磷酸化JNK蛋白增加、磷酸化ERK1／2蛋白下降。结论 C2－神经酰胺可在体外诱导人间皮瘤细胞凋亡，并对其凋亡通路中的关键分子有影响，发挥了一定的抗肿瘤作用。