内源性大麻素诱导人肝癌细胞MHCC97H凋亡中Caspase-3的作用  

Apoptosis induction of anandamide in a human hepatocellular carcinoma cell line with highly metastatic potential（MHCC97H）

作　　者：王艳红[1] 周小芸[1] 张哲[1] 沈沪佳 

机构地区：[1]复旦大学肝癌研究所中山医院,上海200032

出　　处：《中华实验外科杂志》2010年第2期153-155,共3页Chinese Journal of Experimental Surgery

基　　金：教育部留学回国人员科研基金资助项目（200314）

摘　　要：目的 检测内源性大麻素（AEA）是否诱导人肝癌高转移细胞株MHCC97H细胞凋亡、以及Caspase-3在细胞凋亡过程中的作用.方法 分别于AEA（100 μmol/L）作用细胞前后,应用Annexin V Binding方法 、流式细胞仪检测细胞凋亡率;荧光活性检测试剂盒检测细胞Caspase-3活性;半定量逆转录-聚合酶链反应（RT-PCR）方法 ,检测Caspase-3、bcl-2及bax的mRNA表达.结果 AEA作用细胞3、6、12、24 h,细胞凋亡率分别为（14.13±2.17）%、（27.04±3.21）%、（34.98±7.74）%及（81.35±9.71）%,其中24 h的细胞凋亡率与对照组（12.72±0.44）%比较,差异有统计学意义（P〈0.01）.AEA作用细胞24 h,细胞的Caspase-3活性由142.0±5.8增加至226.0±6.4（P〈0.01）;Caspase-3 mRNA的表达由0.24±0.13升高至0.54±0.32（P〈0.05）;如预先加入Caspase-3抑制剂（Z-VAD-FMK）后再加入AEA,则细胞的Caspase-3活性不再升高90.0±4.3.RT-PCR方法 未检测到bcl-2 mRNA及bax mRNA的表达.结论 AEA对人肝癌高转移细胞MHCC97H具有凋亡诱导作用,Caspase-3参与了凋亡的调控.Objective To verify whether anandamide （AEA） has a capacity of inducing apoptosis in a HCC cell line with highly metastatic potential （MHCC97H）, and investigate the role of Caspase-3 in apoptosis. Methods MHCC97H cells were treated with AEA at the different concentrations （0,50 or 100 μmol/L, respectively）, and apoptosis was tested by Annexin V-FITC Apoptosis Detection Kit. Caspase-3 ac-tivity was measured by Caspase-3 Fluorescent Assay Kit, and the Caspase-3 mRNA, bcl-2 and bax were de-tected by reverse transcription polymerase chain reaction （RT-PCR）. Results After treatment with AEA （100 μmol/L） for 24 h, apoptosis rate was （81.35±9.71）%, and Caspase-3 activity in MHCC97-H cells was increased significantly from 142.0±5.8 to 226.0±6.4 （P〈0.01）. There was significant difference in the expression of Caspase-3 mRNA between AEA group （0.54±0.32） and control group （0.24±0.13） following 24 h of treatment （P〈0.01）. The mRNA expression of bcl-2 and bax could not be detected in MHCC97-H cells between AEA group and control group. Conclusion The apoptosis of MHCC97H cells could be induced by AEA via Caspase-3-transducted signal.

关 键 词：癌 肝细胞 脱噬作用 ANANDAMIDE CASPASE-3 

分 类 号：R735.7[医药卫生—肿瘤]