ERK和SAPK/JNK通路在神经酰胺致HT-29细胞凋亡中的作用

doi:10.3969/j.issn.1004616x.2012.06.003

癌变·畸变·突变

ERK和SAPK/JNK通路在神经酰胺致HT-29细胞凋亡中的作用

白艳艳1，2/吴艳萍2/张晓峰2/李百祥2，*

1. 厦门市疾病预防控制中心，福建厦门 361021；2. 哈尔滨医科大学公共卫生学院卫生毒理学教研室，黑龙江哈尔滨 150081

收稿日期:2012-02-06 修回日期:2012-10-16 出版日期:2011-11-30 发布日期:2011-11-30
通讯作者: 李百祥，E-mail：libaix@ems.hrbmu.edu.cn
作者简介:白艳艳(1980- )，女，河北省唐山市人，硕士研究生，理化检验技师，研究方向：食品理化检验。
基金资助:
国家自然科学基金(30471447)

Mechanisms of ERK and SAPK/JNK pathways of ceramide-induced apoptosis in HT-29 cells

BAI Yan-yan1，2，WU Yan-ping2，ZHANG Xiao-feng2，LI Bai-xiang2，*

1. Xiamen Municipal Center for Disease Control and Prevention, Xiamen 361021, Fujian; 2. Department of Health Toxicology, College of Public Health, Harbin Medical University, Harbin 150081, Heilongjiang, China

Received:2012-02-06 Revised:2012-10-16 Online:2011-11-30 Published:2011-11-30
Contact: LI Bai-xiang，E-mail：libaix@ems.hrbmu.edu.cn
About author:白艳艳(1980- )，女，河北省唐山市人，硕士研究生，理化检验技师，研究方向：食品理化检验。
Supported by:
国家自然科学基金(30471447)

摘要/Abstract

摘要：

目的：探讨ERK和SAPK/JNK信号途径在C2-神经酰胺诱导人结肠癌HT-29细胞凋亡中的作用。方法：分别用不同剂量C2-神经酰胺 (0、12.5、25.0和50.0 μmol/L)处理HT-29细胞24 h，分别采用AO/EB染色法观察HT-29细胞凋亡的形态学变化，计算细胞凋亡率；Western Blotting法检测ERK、p-ERK蛋白表达。再分别用不同剂量C2-神经酰胺 (0、12.5、25.0和50.0 μmol/L)处理HT-29细胞24 h，并选择50.0 μmol/L C2-神经酰胺作用3、6、12、24 h后，用SAPK/JNK检测试剂盒结合Western Blotting方法检测phospho-c-Jun蛋白的表达。结果：与溶剂对照组比较，随着C2-神经酰胺剂量的增加，各剂量组HT-29细胞的凋亡率增加 (P＜0.05)，ERK总蛋白和p-ERK的表达呈现下降趋势，存在剂量-效应关系 (P＜0.05)。不同浓度和不同时间的C2-神经酰胺作用下，HT-29细胞中phospho-c-jun的表达无明显变化 (P＞0.05)。结论：C2-神经酰胺能够直接诱导人结肠癌HT-29细胞发生凋亡，在此过程中SAPK/JNK活性无显著变化，抑制ERK信号转导通路可能是其诱导凋亡的机制。

关键词: 神经酰胺, 亡, APK/JNK通路, RK通路

Abstract:

OBJECTIVE: To study the mechanisms of ERK and SAPK/JNK pathways of ceramide-induced apoptosis in HT-29 cells. METHODS：HT-29 cells were treated with C2-ceramide for 24 hours at different doses (0，12.5，25.0，50.0μmol/L). AO/EB fluorescent staining was used to identify the morphological changes of HT-29 cells and evaluate the rate of apoptosis in 200 cells. The protein expressions of ERK and phosphorylated ERK(p-ERK) were evaluated by Western Blotting. The activity of phospho-c-Jun in different doses and treatment time of C2-ceramide groups was assessed using SAPK/JNK assay and Western Blotting. RESULTS：Apoptosis rate was increased and the expressions of ERK and p-ERK were reduced in a dose-dependent manner (P＜0.05). The activity of phospho-c-jun in different doses and treatment time showed no obvious changes (P＜0.05). CONCLUSION：C2-ceramide could directly induce apoptosis in human colon carcinoma HT-29 cells in vitro，and C2-ceramide could inhibit the ERK pathway without activating SAPK/JNK pathway during this process.

Key words: ceramide, poptosis, APK/JNK pathway, RK pathway

白艳艳1，2/吴艳萍2/张晓峰2/李百祥2，*. ERK和SAPK/JNK通路在神经酰胺致HT-29细胞凋亡中的作用[J]. 癌变·畸变·突变, doi: 10.3969/j.issn.1004616x.2012.06.003.

BAI Yan-yan1，2，WU Yan-ping2，ZHANG Xiao-feng2，LI Bai-xiang2，*. Mechanisms of ERK and SAPK/JNK pathways of ceramide-induced apoptosis in HT-29 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004616x.2012.06.003.

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ERK和SAPK/JNK通路在神经酰胺致HT-29细胞凋亡中的作用

Mechanisms of ERK and SAPK/JNK pathways of ceramide-induced apoptosis in HT-29 cells

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