重组人肿瘤坏死因子α联合变异型IL-2治疗肝癌的实验研究  

STUDY OF INHIBITION EFFECT ON RECOMBINATING THERAPYWITH rhTNF-α AND MUTANT IL-2 IN HEPATIC CARCINOMA

作　　者：曹英林[1] 马春红[1] 宋静[1] 张利宁[1] 孙汶生[1] 

机构地区：[1]山东大学医学院微生物学与免疫学教研室

出　　处：《山东医科大学学报》2001年第5期387-390,共4页Acta Academiae Medicinae Shandong

摘　　要：目的 :从大肠杆菌中获得高活性重组人肿瘤坏死α因子 (rhTNF α) ,并探索TNF α与其它细胞因子临床治疗肝癌的合理配伍方案。方法 :CaCl2 法转化大肠杆菌JM 10 9,温度诱导rhTNF α ,收获菌体 ,裂解后 ,Westernblot检测表达产物的特异性 ,薄层分析rhTNF α表达量 ,MTT 短程法测定rhTNF α对L92 9杀伤作用。以表达的rhTNF α作用于肝癌细胞株BEL74 0 2和荷瘤鼠 (肝癌腹水瘤 ) ,验证其对肝癌细胞的体内外抑制作用。采用嵌套设计 ,MTT 短程法测定rhTNF α与变异型IL 2对BEL74 0 2的协同抑制作用。结果 :Westernblot证实 pAW 711转化JM 10 9后筛得的重组菌表达产物特异性良好 ,薄层分析表明rhTNF α表达量可达菌体蛋白的 17 8% ,MTT 短程法证实表达产物具有良好生物活性 ,并可在体外明显杀伤肝癌细胞 ,其杀瘤活性优于标准对照。体内实验证实rhTNF α可使人肝癌腹水瘤荷瘤鼠生命延长率达31 5 %。rhTNF α与变异型IL 2具有协同抑癌作用 ,其协同作用具有特殊的剂量依赖性。结论 :在大肠杆菌中可获得高活性rhTNF α ,rhTNF α在体内外均可明显抑制肝癌细胞 ,变异型IL 2对此有协同作用。Objective:To express recombinant human tumor necrosis factor-α (rhTHF-α) in E.coli,and explore the best way of using rhTHF-α and mutant interleukin-2(IL-2) in hepatic carcinoma therapy.Methods:Bacillas coli JM109 was transformed with CaCl 2,rhTHF-α was expressed after temperature induced.The products were checked by Western blot.The cytotoxicity of rhTHF-α on hepatic carcinoma cell line(BEL7402) and L929 were assayed by MTT.The inhibition of rhTNF-α on hepatic carcinoma cells in vivo was tested.Also the co-inhibition effect of rhTNF-α and mutant IL-2 was measured by MTT.Results:rhTNF-α was checked out by Western blot,the protein can kill L929 cells well.Using MTT assay and animal experiment,cyotoxic effect of rhTNF-α on hepatic carcinoma cells both in vivo and in vitro.Mutant IL-2 can enhance the cytotoxicity of rhTNF-α.Conclusion:rhTNF-α expressed in Bacillas coli JM109 can inhibit the hepatic carcinoma cells,moreover,mutant IL-2 can enhance this effect.

关 键 词：肿瘤坏死因子 白细胞介素2 肝癌 治疗 实验研究 

分 类 号：R735.7[医药卫生—肿瘤] R73－362[医药卫生—临床医学]