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

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（ＲＺ１ ｍｕｔ）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

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

OBJECTIVE: To explore the effect of EBNA-1 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 EBNA-1 ribozyme (RZ1) or its inactive mutant (RZ1mut) were constructed as Ad.RZ1 or Ad.RZ1mut. 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 (1 x 10(7) 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 EBNA-1 ribozyme and EBNA-1 mRNA in the tumor was analyzed by RT-PCR, and the expression of EBNA-1 protein in the tumor was detected by Western blot. RESULTS: The tumors (0.27 +/- 0.18) g formed from the Ad.RZ1-treated LCLs were much smaller than those of the untreated LCLs (1.04 +/- 0.27) g or LCLs treated with control vector of adenovirus (1.12 +/- 0.32) g (P < 0.01), and also smaller than those of Ad.RZ1mut-treated LCLs (0.76 +/- 0.28) g (P < 0.05). The expression of EBNA-1 mRNA and protein in the tumors of Ad.RZ1-treated LCLs decreased significantly than that of other groups. CONCLUSIONS: Adenovirus delivery of EBNA-1 ribozyme was able to suppress LCL tumors significantly and depress the tumor genesis ability of B lymphocyte induced by EBV.