纳米脂质体介导酪氨酸激酶受体3配体基因转染树突状细胞对结肠癌细胞凋亡的作用  

Effect of nano-liposome-mediated FL gene transfection on apoptosis of colon cancer cells

作　　者：孙念峰[1] 孟庆义[1] 田爱玲[1] 王瑞华[1] 刘兆轩[1] 胥莉[1] 

机构地区：[1]山东大学附属济南市中心医院普外科,济南250013

出　　处：《中华实验外科杂志》2011年第11期1912-1914,共3页Chinese Journal of Experimental Surgery

基　　金：教育部新教师基金资助项目（200804221081）;山东省自然科学基金资助项目（Y2008C08）;中国博士后科学基金资助项目（20100481271）

摘　　要：目的观察利用阳性纳米脂质体介导的基因治疗联合免疫治疗对结肠癌细胞的治疗作用。方法应用基因克隆技术分别构建含有绿色荧光蛋白报告基因的重组质粒（pEGFP），并依据静电吸附原理制备携带基因的阳离子纳米脂质体，再将转染阳性的树突状细胞（DC）移植作用于结肠癌细胞。结果成功构建含有绿色荧光蛋白报告基因的重组质粒，并得到酶切鉴定和序列测定证实。携带酪氨酸激酶受体3配体（FL）基因的阳离子纳米脂质体成功转染DC，发现基因在细胞内得到很好表达，且随着培养时间的增加，蛋白表达量有所增加，72h时表达量最高。基因移植后发现对照组肿瘤体积增长最快，FL基因组肿瘤体积增长较慢；FL基因组肿瘤细胞凋亡率和死亡率明显高于对照组（P〈0．05）；而pEGFP组和对照组差异无统计学意义（P〉0．05）。结论阳离子纳米脂质体作为基因载体具有一定优越性。基因治疗联合免疫治疗在结肠癌治疗中具有重要意义。Objective To investigate the therapeutic effects of cationic nano-liposome-mediated gene therapy combined with immunotherapy for colon cancer. Methods Recombinant plasmids containing green and red fluorescent protein reporter genes were constructed using gene cloning methods. Gene-carrying cationic nano-liposomes were prepared based on the electrostatic adherence principle and then transfected into dendritic cells （DC） that were transplanted into colon cancer cells. Results Recombinant plasmids containing green or red fluorescent protein reporter genes were successfully constructed using gene cloning methods and confirmed by restriction enzyme digestion and sequencing. Gene-carrying cationic nano-liposomes were transfected into colon cancer cells, and gene expression was detected. The apoptosis and cell death was observed in the tyrosine kinase receptor 3 ligand （FL） group, and the lowest level was detected in the control group. The results revealed significantly higher level of apoptisis and lower level of proliferation in the FL group than in the control groups （ P 〈 0. 05）. However, the lowest level of tumor cell apoptisis and the highest level of cell proliferation were observed in the control group. Conclusion Cationic nano-liposomes have the advantages of being gene carriers. Gene therapy combined with immunotherapy may be important for colon cancer.

关 键 词：阳性纳米脂质体 FL基因 基因治疗 结肠癌 

分 类 号：R735.35[医药卫生—肿瘤]