实用肿瘤学杂志 ›› 2014, Vol. 28 ›› Issue (6): 492-497.doi: 10.11904/j.issn.1002-3070.2014.06.003

• 论著 • 上一篇    下一篇

C2-神经酰胺对人间皮瘤细胞凋亡的诱导作用

张天华1, 李永春2, 李宪伟1, 迟德财1   

  1. 1.哈尔滨医科大学附属第二医院普外五科(哈尔滨 150086);
    2.牡丹江医学院红旗医院普外科
  • 收稿日期:2014-03-29 出版日期:2014-12-28 发布日期:2015-01-08
  • 通讯作者: 张天华,E-mail:hunterzth@163.com
  • 作者简介:张天华,男,(1977-),博士,副主任医师,从事肿瘤凋亡与防治的研究
  • 基金资助:

    黑龙江省留学归国科学基金项目(LC2009C17)

C2-ceramide induced apoptosis of malignant mesothelioma cells in vitro

ZHANG Tianhua1,LI Yongchun2,LI Xianwei1,CHI Decai1   

  1. 1.Department of General Surgery,The Second Affiliated Hospital of Harbin Medical University,Harbin 150086,China;
    2.Department of General Surgery,Hongqi Hospital of Mudanjiang
  • Received:2014-03-29 Online:2014-12-28 Published:2015-01-08

摘要:

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。

关键词: 神经酰胺, 间皮瘤, 细胞凋亡

Abstract:

Objective In this research,the effects of exogenous C2-ceramide on the induction of apoptosis of mesothelioma cells in vitro and related important proteins are investigated.Methods Mesothelioma cells were treated with various doses of C2-ceramide for different duration.Cell viability were analyzed by cell counting 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 significantly 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 ERK1/2 kinase was decreased.Conclusion Our results indicate that C2-ceramide induced apoptosis of malignant mesothelioma cells in vitro.This anti-tumor affect is achieved by adjusting related important proteins.

Key words: Ceramide, Mesothelioma, Apoptosis

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