甘露糖敏感性绿脓杆菌制剂诱导肝细胞癌凋亡的机制研究  

The mechanism of mannose sensitive hamemagglutination vaccine in inducing apoptosis of hepatocellular carcinoma

作　　者：李涛[1] 汤钊猷[2] 周建炜[2] 樊嘉[2] 胡三元[1] 智绪亭[1] 

机构地区：[1]山东大学齐鲁医院普外科,济南250012 [2]复旦大学附属中山医院肝癌研究所,上海

出　　处：《中华肝胆外科杂志》2011年第10期838-841,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然基金资助（30901444,30972889）

摘　　要：目的探讨甘露糖敏感性绿脓杆菌制剂（pseudomonasaeruginosa marnose sensitive hamemagglutination vaccine, PA-MSHA）诱导肝细胞癌凋亡的机制。方法将具有肺转移潜能的人MHCC97L肝癌组织块种植于BAI，B／enu／nu雄性裸鼠肝脏，建立转移性人肝癌裸鼠原位模型。荷瘤裸鼠随机分为对照组、PA—MsHA腹腔注射组及皮下注射组。各组均于种植后第3天开始用药，于种植后第6周末处死动物。ELISA法检测血清TNF-a、IL-4、IL6、IFN-γ水平，分光光度法检测肿瘤组织裂解液中Caspase3、Caspase8、Caspase9的酶活性，Western Blot检测肿瘤组织内Fas和FasL蛋白水平。结果对照组、PA—MSHA皮下注射组和腹腔注射组裸鼠血清TNF—a分别为（25．24±3．22）pg／ml、（25．50±4．55）pg／ml（P〉0．05）和（34．22±2．42）pg／ml（P〈0．01）。三组血清IL-4、IL-6及IFN-γ水平无显著差异（P〉0．01）。腹腔注射组肿瘤组织裂解液中Caspase3、Caspase8、Caspase9的酶活性比对照组分别上调4．1倍、2．3信和1．9倍（P〈0．01）。与对照组比较，PA—MSHA皮下注射组和腹腔注射组肝癌组织内Fas和FasL蛋白的表达显著增加。结论PA—MSHA腹腔注射可通过上调TNF—a及Fas／FasL表达诱导肝细胞肝癌的凋亡，从而抑制其生长和转移。Objective To investigate the mechanism of pseudomonasaeruginosa mannose sensitive hamemagglutination vaccine （PA-MSHA） in inducing apoptosis in hepatocellular carcinoma （HCC）. Methods A metastatic model of human hepatocellular carcinoma （HCC） was established by orthotopic implantation of histologically intact human HCC tissue into the liver of nude mice. Mice bearing xenografts in liver were randomly divided into three groups： control group, PA-MSHA intrap-eritoneal administration group, and PA MSHA subcutaneous administration group. The agent was ad- ministered every day after the third day post tumor implantation. At the end of the sixth week, the mice were killed. Serum levels of TNF-a, IL-4, IL-6 and IFN y were measured by ELISA and the activities of caspase 3, caspase 8 and caspase 9 in the tumor samples were tested by spectrophotometrie method. Fas/FasL expressions were evaluated by Western blotting. Results Serum TNF a levels in the control group, PA MSHA subcutaneous administration group and PA-MSHA intraperitoneal administration group were 25.2±3.22 pg/ml, 25.50±4.55 pg/ml（P〉0. 05） and 34.22±2.42 pg/ml （P〈0.01）, respectively, while there were no significant differences in serum IL-4, IL-6 and IFN-γ among these three groups（P〉0.01）. Compared with the control group, the activities of caspase 3, caspase 8 and easpase 9 in tumors in the intraperitoneal administration group increased by 4.1 , 2.3 and 1.9 fold（P〈0.01）, respectively, and Fas/FasL expressions were significantly elevated in the intraperitoneal administration group and subcutaneous administration group. Conclusion When given intraperitoneally, PA MSHA can induce apoptosis in HCC by promoting the secretion of TNF-a and expression of Fas/FasL, thereby inhibiting HCC growth and metastasis.

关 键 词：癌 肝细胞 绿脓杆菌制剂 凋亡 肿瘤坏死因子-A 

分 类 号：R735.7[医药卫生—肿瘤]