nHAP-Mg2+纳米颗粒对肝癌HepG2细胞的基因转染的影响  

Influence of nHAP-Mg2+ nanoparticle on the gene transferring of hepatoma cell line HepG2

作　　者：李高鹏[1] 叶露 潘景升[1] 邓宝雯[1] 马健鸿[1] 王浩虎[1] 罗葆明[1] 

机构地区：[1]中山大学孙逸仙纪念医院超声科,广东广州510120 [2]广州市第八医院传染科,广东广州510230

出　　处：《岭南现代临床外科》2012年第2期99-102,共4页Lingnan Modern Clinics in Surgery

基　　金：广东省医学科研课题(B2011094);广东省科技计划社会发展(2010B031100009)

摘　　要：目的研究新型非病毒载体羟基磷灰石纳米粒子(hydroxyapatite nanoparticle,nHAP)在肿瘤基因治疗中的作用及机制。方法 1)应用氯化镁修饰nHAP,以增强型绿色荧光蛋白基因(PEGFP-N1)为报告基因,联合脂质体共同作为基因载体转染肝癌细胞HepG2,流式细胞术测定转染效率和平均荧光值;2)琼脂糖凝胶电泳研究nHAP-Mg2+与DNA的结合和保护作用;3)MTT法评价nHAP-Mg2+对细胞生长的影响。结果 1)nHAP-Mg2+单独无明显基因转移功能,联合脂质体后,可成功将绿色荧光蛋白基因转染进入HepG2肝癌细胞,其转染的平均荧光的强度和转染的效率较均单用应用脂质体组显著提高(P<0.05);2)在合适的比例时nHAP-Mg2+与DNA能够完全结合并能够耐受DNA酶的破坏消化从而起到保护作用;3)应用单纯nHAP-Mg2+或联合脂质体时,随着量的增加对细胞的生长抑制增强(P<0.05)。结论 nHAP-Mg2+具有结合和保护质粒DNA的作用。在基因治疗时,可以作为脂质体的辅助载体应用,不但可以提高转染效率和稳定表达,还可增强基因治疗的抗肿瘤效果,适用于肿瘤的基因治疗。Objective To study the role of hydroxyapatite nanoparticle in the gene transferring of hepatoma cell line HepG2 and the possible mechanisms. Methods ①Throμgh combining Magnesium chloride modified hydroxyapatite nanoparticles （nHAP） and liposome,the PEGFP-N1 plasmids were successfully transfected into HepG2 cells. Both the rate of gene transfering and the related mean fluorescence intensity were observed and analyzed by flow cytometry. ②The combination and protection of nHAP-Mg2＋ to DNA were analyzed by gelose gelatin electrophoresis. ③The effect nHAP-Mg2＋ to the growth of the cells were studied by MTT. Results ①The gene was not transferred by nHAP-Mg2＋ alone. Throμgh combining liposome and the nanoparticles, the green fluorescence protein gene was transferred extremely efficiently and its transfering rates were significantly higher than the liposome group（P0.05）. ②At certain proportion, nHAP-Mg2＋ can combine the plasmids compeletly and protected the DNA from digestion of nucleinase. ③The inhibition of cell growth were along with the concentration of nHAP-Mg2＋ whether it was used alone or with the combination of lipid（P0.05）. Conclusions nHAP-Mg2＋ has the ability of combining and protecting DNA and can be used to transfer gene as the adjunct carrier of lipid for the gene therapy of tumor cells to elevate the gene tansfering and expression rate and also enhance the anti-tumor effection.

关 键 词：羟基磷灰石纳米粒子 肝癌 作用机制 基因治疗 

分 类 号：R735.3[医药卫生—肿瘤]