EB病毒核抗原-1特异性核酶对成淋巴细胞系肿瘤形成能力的影响  

The effect of EBNA1 ribozyme on the ability of tumor genesis of lymphoblastoid cell lines in SCID mice

作　　者：刘爱华[1] 姜勇 赵克森 黄爽 

机构地区：[1]第一军医大学南方医院呼吸科

出　　处：《中华医学杂志》1999年第7期538-541,共4页National Medical Journal of China

基　　金：国家自然科学基金

摘　　要：目的探讨腺病毒载体介导的ＥＢ病毒核抗原１（ＥＢＮＡ１）特异性核酶对成淋巴细胞系（ＬＣＬｓ）在严重的联合免疫缺陷（ＳＣＩＤ）小鼠体内致瘤性的影响。方法用分子克隆技术构建ＥＢＮＡ１核酶（ＲＺ１）及其无活性诱变体（ＲＺ１ｍｕｔ）的重组腺病毒表达载体（ＡｄＲＺ１和ＡｄＲＺ１ｍｕｔ）。通过同源重组方式在２９３细胞生成腺病毒，并通过噬斑形成进行分离建立ＬＣＬ。将感染重组腺病毒的ＬＣＬ细胞注射于小鼠颈背部皮下，１０～１４天后形成明显的肿块，６周后观察剔除的肿瘤大小和重量。用ＲＴＰＣＲ法分析肿瘤组织ＥＢＮＡ１核酶和ＥＢＮＡ１ｍＲＮＡ的表达。用Ｗｅｓｔｅｒｎ印迹法分析肿瘤组织内ＥＢＮＡ１蛋白的表达。结果用ＡｄＲＺ１处理的ＬＣＬ细胞形成的肿瘤块（０２７±０１８）ｇ明显小于对照组（１０４±０２７）ｇ和腺病毒对照组（空载体）的肿瘤块（１１２±０３２）ｇ（Ｐ＜００１），也明显小于ＡｄＲＺ１ｍｕｔ组的肿瘤块（０７６±０２８）ｇ（Ｐ＜００５）。ＡｄＲＺ１组肿瘤组织ＥＢＮＡ１ｍＲＮＡ和蛋白的表达也较其他组明显减少。结论腺病毒输送的ＥＢＮＡ１核酶能够抑制ＬＣＬ细胞在体内的增殖，降低ＥＢ病毒诱导?Objective To explore the effect of EBNA1 specific ribozyme on the tumor genesis ability of lymphoblastoid cell lines (LCLs) in severe combined immunodeficiency (SCID) mice Methods Using molecular cloning technique, recombinant adenovirus vectors expressing EBNA1 ribozyme (RZ1) or its inactive mutant (RZ1mut) were constructed as AdRZ1 or AdRZ1mut Recombinant adenoviruses were generated by homologous recombination and isolated by plaque formation LCLs were established and infected with recombinant adenovirus and then injected in SCID mice (1107 cells/animal) The SCID mice were sacrificed six weeks after injection, then the tumors were excised from these mice and their size and weight were measured The expression of EBNA1 ribozyme and EBNA1 mRNA in the tumor was analyzed by RTPCR, and the expression of EBNA1 protein in the tumor was detected by Western blot Results The tumors (027018) g formed from the AdRZ1treated LCLs were much smaller than those of the untreated LCLs(104027)g or LCLs treated with control vector of adenovirus (112032) g (P<001), and also smaller than those of AdRZ1muttreated LCLs(076028) g (P<005) The expression of EBNA1 mRNA and protein in the tumors of AdRZ1treated LCLs decreased significantly than that of other groups Conclusions Adenovirus delivery of EBNA1 ribozyme was able to suppress LCL tumors significantly and depress the tumor genesis ability of B lymphocyte induced by EBV

关 键 词：EB病毒核抗原1 特异性核酶 成淋巴细胞瘤 肿瘤 

分 类 号：R730.231.3[医药卫生—肿瘤]