BikDD肿瘤靶向基因治疗抑制淋巴瘤细胞生长研究  

Tumor-Targeted BikDD Gene Therapy Inhibits Lymphoma Cells Growth

作　　者：曹军[1] 罗宁[2] 李建璜[2] 

机构地区：[1]中南大学湘雅医学院附属肿瘤医院胸部内二科,长沙410013 [2]中南大学湘雅医院肿瘤科,长沙410008

出　　处：《成都医学院学报》2015年第1期16-20,共5页Journal of Chengdu Medical College

基　　金：湖南省自然科学基金课题资助(NO:09jj5016)

摘　　要：目的构建含突变型Bik(BikDD)、Survivin启动子及VP16-GAL4-WPRE整合型系统性放大系统(VISA)的淋巴瘤基因治疗载体S-VISA-BikDD,研究其在淋巴瘤细胞中抑制细胞生长的作用机制。方法应用基因重组技术构建载体S-VISA-BikDD,用电转染法将其转入淋巴瘤细胞株Ramos、Raji和U937,Western blot检测转染前后Bik蛋白表达,通过流式细胞仪检测Raji细胞凋亡情况,MTT实验观察细胞生长抑制状况。结果通过PCR法及酶切鉴定证明载体S-VISA-BikDD构建成功,转染后淋巴瘤细胞Bik表达水平有所提高,同时可促进淋巴瘤细胞凋亡,并抑制其生长。结论 S-VISA-BikDD是一种有潜力的淋巴瘤基因治疗载体,S-VISA-BikDD载体的成功构建及对其促凋亡作用的研究,为淋巴瘤基因治疗找到了新的思路及靶标,具有重要的理论意义和应用前景。Objective To construct a Lymphoma specific targeted gene therapy vector S-VISA-BikDD, composed of mutant Bik（BikDD）, the survivin promoter,and the VP16-GAL4-WPRE integrated systemic amplifier system（VISA）, to investigate its role of growth inhibition in Lymphoma cell lines. Methods S-VISA-BikDD was constructed by the way of gene recombination technology. The lymphoma ceil lines Raji, Ramos and U937 were transfected with S-VISA-BikDD. Western blot was used to assess the different expression of Bik in lymphoma ceil lines before and after the transfection was conducted. The flow cytometry was carried out to detect pro apoptotic effects and the MTT assay was carried out to detect growth inhibition in lymphoma cell lines. Results S-VISA- BikDD was proved to be constructed correctly by PCR and enzyme digestion. The expression level of BikDD in lymphoma cells was enhanced after the transfection was performed. It enhanced cell apoptosis and inhibited cell proliferation in lymphoma cells. Conclusion S-VISA BikDD is a potent Lymphoma Gene Therapy vector, which provides a new method and target for further study of lymphoma gene therapy.

关 键 词：BikDD SURVIVIN启动子 VISA S-VISA-BikDD 淋巴瘤 

分 类 号：R733.1[医药卫生—肿瘤]