JNK/SAPK信号转导系统在三氧化二砷诱导肝癌细胞凋亡中的作用
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李航宇, 钟鑫平, 隋春阳, 孔凡民, 董 明, 张 浩, 刘金钢
三氧化二砷;肝癌;MTT法;流式细胞术;Westernblot法,李航宇,刘金钢,钟鑫平,隋春阳,孔凡民,董明,张浩,李航宇,通讯作者:,RoleofJNK,SPARKsignaltransductionsystemi
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李航宇, 刘金钢, 中国医科大学附属第二医院普外科 辽宁省沈阳市 110004
钟鑫平, 第二军医大学附属东方肝胆医院胆道二科 上海市 200433
隋春阳, 孔凡民, 董明, 张浩, 中国医科大学附属第一医院普外科 辽宁省沈阳市 110001
李航宇, 医学博士, 讲师, 主治医师, 主要从事肝胆恶性疾病的基础与临床研究.
国家自然基金资助项目, No. 30572114
通讯作者: 李航宇, 110004, 辽宁省沈阳市, 中国医科大学附属第二医院普外科. li_hangyu@126.com
电话: 024-83955279
收稿日期: 2006-12-20 接受日期: 2007-01-10
Role of JNK/SPARK signal transduction system in the apoptosis of hepatocellular carcinoma cell line HepG2 induced by arsenic trioxide
Hang-Yu Li, Xin-Ping Zhong, Chun-Yang Sui, Fan-Min Kong, Ming Dong, Hao Zhang, Jin-Gang Liu
Hang-Yu Li, Jin-Gang Liu,Department of General Surgery, the Second Affiliated Hospital of China Medical University, Shenyang 110004, Liaoning Province, China
Xin-Ping Zhong,the Second Department of Biliary Tract Surgery, Dongfang Hepatobilliary Hospital Affiliated to the Second Military Medical University, Shanghai 200433, China
Chun-Yang Sui, Fan-Min Kong, Ming Dong, Hao Zhang,Department of General Surgery, the First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
Supported byNation Natural Science Foundation of China, No. 30572114
Correspondence to:Hang-Yu Li, Department of General Surgery, the Second Affiliated Hospital of China Medical University, Shenyang 110004, Liaoning Province, China. li_hangyu@126.com
Received:2006-12-20 Accepted:2007-01-10
Abstract
AIM: To explore of the role of JNK/SAPK signal transduction system in the apoptosis of human hepatocellular carcinoma cell line HepG2 induced by arsenic trioxide (As2O3).
METHODS: Human hepatocellular carcinoma cell line HepG2 was treated with different concentrations of As2O3. MTT assay was used to observe the proliferation of HepG2 cells, and flow cytometry was used to examine the apoptosis rate and growth cycles of the cells. The expression of p-MEK4, JNK, p-JNK, Caspase-3 and poly (ADP ribose) polymerase (PARP) were detected by Western blot before and after JNK signal was blocked by SP600125, a JNK inhibitor.
RESULTS: As2O3 inhibited the proliferation of HepG2 cells in a concentration- and time-dependent manner. Flow cytometry showed that As2O3 induced the apoptosis of HepG2 cells in a time-dependent way, and the cells were mainly blocked in G2/M phase (7.22% ± 1.50%, 11.56% ± 0.73%, 33.8% ± 1.62%, and 46.02% ± 0.11% for 0, 24, 48, and 72 hours, respectively). Western blotting exhibited that As2O3 induced the activation of Caspase-3 and PARP cleavage. The expression of p-MEK4 and p-JNK were detected 10 minutes after As2O3 treatment, reached the peak at the 20th minute, but decreased gradually from the 30th minute, and preceded apoptosis ......
李航宇, 刘金钢, 中国医科大学附属第二医院普外科 辽宁省沈阳市 110004
钟鑫平, 第二军医大学附属东方肝胆医院胆道二科 上海市 200433
隋春阳, 孔凡民, 董明, 张浩, 中国医科大学附属第一医院普外科 辽宁省沈阳市 110001
李航宇, 医学博士, 讲师, 主治医师, 主要从事肝胆恶性疾病的基础与临床研究.
国家自然基金资助项目, No. 30572114
通讯作者: 李航宇, 110004, 辽宁省沈阳市, 中国医科大学附属第二医院普外科. li_hangyu@126.com
电话: 024-83955279
收稿日期: 2006-12-20 接受日期: 2007-01-10
Role of JNK/SPARK signal transduction system in the apoptosis of hepatocellular carcinoma cell line HepG2 induced by arsenic trioxide
Hang-Yu Li, Xin-Ping Zhong, Chun-Yang Sui, Fan-Min Kong, Ming Dong, Hao Zhang, Jin-Gang Liu
Hang-Yu Li, Jin-Gang Liu,Department of General Surgery, the Second Affiliated Hospital of China Medical University, Shenyang 110004, Liaoning Province, China
Xin-Ping Zhong,the Second Department of Biliary Tract Surgery, Dongfang Hepatobilliary Hospital Affiliated to the Second Military Medical University, Shanghai 200433, China
Chun-Yang Sui, Fan-Min Kong, Ming Dong, Hao Zhang,Department of General Surgery, the First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China
Supported byNation Natural Science Foundation of China, No. 30572114
Correspondence to:Hang-Yu Li, Department of General Surgery, the Second Affiliated Hospital of China Medical University, Shenyang 110004, Liaoning Province, China. li_hangyu@126.com
Received:2006-12-20 Accepted:2007-01-10
Abstract
AIM: To explore of the role of JNK/SAPK signal transduction system in the apoptosis of human hepatocellular carcinoma cell line HepG2 induced by arsenic trioxide (As2O3).
METHODS: Human hepatocellular carcinoma cell line HepG2 was treated with different concentrations of As2O3. MTT assay was used to observe the proliferation of HepG2 cells, and flow cytometry was used to examine the apoptosis rate and growth cycles of the cells. The expression of p-MEK4, JNK, p-JNK, Caspase-3 and poly (ADP ribose) polymerase (PARP) were detected by Western blot before and after JNK signal was blocked by SP600125, a JNK inhibitor.
RESULTS: As2O3 inhibited the proliferation of HepG2 cells in a concentration- and time-dependent manner. Flow cytometry showed that As2O3 induced the apoptosis of HepG2 cells in a time-dependent way, and the cells were mainly blocked in G2/M phase (7.22% ± 1.50%, 11.56% ± 0.73%, 33.8% ± 1.62%, and 46.02% ± 0.11% for 0, 24, 48, and 72 hours, respectively). Western blotting exhibited that As2O3 induced the activation of Caspase-3 and PARP cleavage. The expression of p-MEK4 and p-JNK were detected 10 minutes after As2O3 treatment, reached the peak at the 20th minute, but decreased gradually from the 30th minute, and preceded apoptosis ......
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