摘要
目的:包装重组分泌型内皮抑素腺相关病毒(rAAV-Endostatin)制备内皮抑素原位基因治疗膀胱癌的基因载体药物。方法:采用分子生物学方法构建腺相关病毒质粒载体(pAAV-IgG-Endostatin),酶切、测序鉴定;经质粒共转染方法包装rAAV-Endostatin,测定病毒颗粒滴度并电镜下鉴定;转染膀胱癌EJ细胞后,ELISA法测定重组内皮抑素的浓度。结果:pAAV-IgG-Endostatin的构建和rAAV-Endostatin的包装均获成功,病毒滴度达1.0×1012v.p/ml。感染EJ细胞后内皮抑素成功表达并分泌,上清液中浓度达54.09ng/ml。结论:rAAV-Endostatin载体的构建、包装与表达的成功为rAAV-Endostatin原位基因治疗膀胱癌的实验研究奠定了基础。
Objective： To package rAAV-Endostatin for endostatin gene therapy of bladder cancer. Methods： To construct pAAV-IgG-Endostatin and identified by enzyme digesting and sequencing method, rAAV-Endostatin was packaged by co-transfection technique then identified under electronic microscope and assayed by ELISA. EJ cells were transfected by rAAV-Endostatin and endostatin concentration in supernatant was assayed by ELISA too. Results： pAAV-IgG-Endostatin was constructed and rAAV-Endostatin was packaged successfully, virus titer is 1.0×10^12 v.p/ml, virus pattern is normal. After EJ cells being transfected, endostatin was expressed and secreted, endostatin concentration in the supernatant is 54.09 ng/ml. Conclusion： rAAV-Endostatin was packaged successfully and thus endostatin gene therapy of bladder cancer was made possible.
出处
《天津医科大学学报》
2006年第1期24-26,29,共4页
Journal of Tianjin Medical University
基金
天津市高等学校科技发展基金资助(20040227)
