胆固醇衍生物阳离子脂质体复合EGFP-IL-1Ra质粒体外转染HELA细胞系的研究(1)
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[摘要] 目的 研究胆固醇衍生物阳离子脂质体复合EGFP-IL-1Ra质粒体外转染HELA细胞系的效率，为类风湿性关节炎治疗提供新的选择。 方法 制备新型胆固醇衍生物脂质体及EGFP-IL-1Ra质粒，两者结合转染HELA细胞系，通过体外凝胶阻滞试验检测阳离子脂质体与质粒DNA的最佳配比关系，MTT法检测脂质体细胞毒性，荧光免疫法检测HELA细胞的转染效率。 结果 实验中所用脂质体的IC50 为650 μg/mL，较对照组有更低的细胞毒性。脂质体/DNA 质量比为50∶1作为最适比例，荧光显微镜观测脂质体结合EGFP-IL-1Ra质粒转染 HELA荧光强度高于单纯EGFP-IL-1Ra质粒转染组。 结论 胆固醇衍生物阳离子脂质体复合EGFP-IL-1Ra质粒可高效的转染HELA细胞系，有望成为类风湿性关节炎治疗的新方法。
[关键词] 胆固醇衍生物；阳离子脂质体；HELA细胞系；类风湿性关节炎
[中图分类号] R-33 [文献标识码] C [文章编号] 1674-4721(2012)07(a)-0005-03
The transgene effect of cholesterol derivative-cationic liposome EGFP-IL-1Ra plasmid complex in HELA cell line
CAI Wenhong1 SUN Baodong1 ZHANG Baofeng2 CHENG Wenxiang3 YUE Ye3 HU Yiping3 LI Jinchao3 ZHANG Peng3
1.Department of Rheumatology, Shenzhen People′s Hospital of Guangdong Province, Shenzhen 518020, China; 2.Department of Nutrition, Shenzhen People′s Hospital of Guangdong Province, Shenzhen 518039, China; 3.Center for Translational Medicine R & D, Shenzhen Institutes of Advanced Technology, Chinese Academy of Science, Guangdong Province, Shenzhen 518055, China
[Abstract] Objective To observe the transgene effect of cholesterol derived cation liposome combine EGFP-IL-1Ra plasmid in HELA cell line, exploring new therapeutic methods for the treatment of rheumatoid arthritis. Methods Cholesterol derived cation liposome as well as EGFP-IL-1Ra plasmid were prepared and combined to transfer the HELA cell line. Gel retarding was performed to select the appropriate liposome/DNA ratio, MTT was used to observe the cell toxicity of this complex.The transfer efficiency was observed using fluorescence microscope ......
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