阳离子脂质体——病毒增强质粒复合物在肝细胞癌基因治疗中的应用

目的建立一种简便有效的非病毒基因治疗方法。方法用构建的含人ＩＬ┐２基因的腺相关病毒（ＡＡＶ）增强质粒（ｐＡＩ┐ｈＩＬ┐２）与新型阳离子脂质体（Ｄｏｓｐｅｒ）以适当比例混合形成复合物（Ｄｏｓｐｅｒ┐ｐＡＩ┐ｈＩＬ┐２）。体外转染小鼠肝癌细胞系ＭＭ４５Ｔ．Ｌｉ┐２，评估了人ＩＬ┐２基因表达情况。应用直接瘤体内注射方法评估了此方法介导ＩＬ┐２基因的抗肿瘤作用。结果应用Ｄｏｓｐｅｒ┐ｐＡＩ┐ｈＩＬ┐２复合物体外转染ＭＭ４５Ｔ·Ｌｉ，其ＩＬ┐２表达水平每毫升可达１７５Ｕ／４８小时，最高表达时间在转染后第４天（２２５Ｕ／ｍｌ），第１２天仍可表达３０Ｕ／ｍｌ。瘤体内直接注射Ｄｏｓｐｅｒ┐ｐＡＩ┐ｈＩＬ┐２复合物后，瘤组织内可检出ＩＬ┐２ｍＲＮＡ，治疗组肿瘤生长明显减慢，小鼠生存率（转染后３０天）明显提高（Ｐ＜０．０１）。结论Ｄｏｓｐｅｒ┐ＡＡＶ增强质粒复合物能有效介导ＩＬ┐２基因表达，产生较好的抗瘤效应，此方法简便、更适合于临床应用

APPLICATION OF CATIONIC LIPOSOME ADENO ASSOCIATED VIRUS BASED PLASMID COMPLEXES FOR GENE THERAPY OF HEPATOCELLULAR CANCER

Objective:To build a simplified efficient non virus method for gene therapy.Methods:Dosper DNA complexes were formed by mixing Dosper with adeno associated virus based plasmid(pAI hIL 2)at appropriate proportion.Human IL 2 gene expression was evaluated in MM45T.Li in vitro.Antitumor activity of Dosper pAI hIL 2 complexes was evaluated in a hepatocellular cancer model in mice with direct intratumor injection.Results:In vitro transfection of MM45T.Li with Dosper pAI hIL 2 complexes resulted in the expression of 175 U of biologically active IL2/ml.The IL2 expression reached peak level at 4 days after transfection(225 U/ml) and persisted for at least 12 days(30 U/ml).The intratumor injection of Dosper pAI hIL 2 complexes resulted in the expression of mRNA,significant reduction in tumor volume and significant increase in the percentage of surviving mice 30 days after transfection.Conclusion:Dosper AAV based plasmid complexes can achieve an efficient IL2 gene expression and induce potent antitumor effect. This method is simple and suitable for clinical cancer therapy.