摘要
目的 探讨新型非病毒载体羟基磷灰石纳米粒子（nHAP）在肿瘤基因治疗中的作用及可能机制.方法 nHAP经氯化镁修饰后,经琼脂糖凝胶电泳判断nHAP-Mg2＋与DNA的结合和保护作用.以增强型绿色荧光蛋白真核表达载体PEGFP-N1为报告基因,以脂质体作为基因载体转染到人类结肠癌SW480/M5细胞,流式细胞术测定转染效率和平均荧光值.四甲基偶氮唑蓝（MTT）比色法评价nHAP-Mg2＋对细胞生长的影响.结果 在质量比合适时,nHAP-Mg2＋与DNA能够完全结合并对DNA起到保护作用.单独应用nHAP-Mg2＋时无基因转移功能,联合脂质体共同作为基因转染的载体可成功将PEGFP-N1基因导入SW480/M5细胞,其转染效率和平均荧光强度高于单用脂质体组（P＜0.05）.单纯应用nHAP-Mg2＋和联合脂质体时,随着nHAP-Mg2＋量的增加（0 ～ 50 μg/ml）对SW480/M5细胞的生长抑制作用增强（P＜0.05）.结论 nHAP-Mg2＋具有结合和保护质粒DNA的作用.作为脂质体的辅助载体,不但可以提高基因转染效率和稳定表达,还可增强基因治疗的抗肿瘤效果.
Objective To investigate the role of hydroxyapatite nanoparticle （nHAP） in the gene transfection of human colorectal cancer cell line SW480/M5 and the possible mechanisms.Methods The combination and protection of nHAP-Mg2＋ to DNA were analyzed by gelose gelatin electrophoresis.Liposome and nHAP modified by magnesium chloride was combined,and the PEGFP-N1 plasmids were transfected into SW480/M5 cells.The gene transfection rate and the mean fluorescence intensity were observed by flow cytometry.The effect of nHAP-Mg2＋ on the growth of the cells were studied by MTT.Results At appropriate proportion,nHAP-Mg2＋ could combine the plasmids compeletly and protected the DNA.The gene could not be transferred by nHAP-Mg2＋ alone.Combining the nanoparticles and liposome,the gene could be transferred very efficiently and the transfection rates were significantly higher than the liposome （P ＜ 0.05）.The inhibition of cell growth was increased along with the concentration of nHAP-Mg2＋ wether it was used alone or with the combination of liposome （P ＜ 0.05）.Conclusions nHAP-Mg2＋ has the ability to combining and protecting DNA and can be used to transfer gene as the adjunct carrier of liposome for the gene therapy of tumor cells to elevate the gene tansfection and expression rate and also enhance the anti-tumor effection.
出处
《肿瘤研究与临床》
CAS
2013年第12期809-811,共3页
Cancer Research and Clinic
关键词
羟基磷灰石类
纳米粒
转染
结肠肿瘤
基因疗法
Hydroxyapatites
Nanospheres
Transfection
Colonic neoplasms
Gene therapy
