硅纳米载体携带RNA干扰质粒逆转人结肠癌细胞多药耐药的实验研究  

Study of Silica Nanoparticles Carrying the Vector of RNAi in the Reversal of Multidrug Resistance of Colorectal Cancer Cell Line

作　　者：姜习新 杨昊 杨毅 

机构地区：[1]湖南省岳阳市二人民医院,湖南岳阳414000

出　　处：《实用预防医学》2013年第11期1385-1389,共5页Practical Preventive Medicine

基　　金：湖南省科技厅资助项目(2009FJ3115)

摘　　要：目的探讨硅纳米颗粒作为基因转染载体携带干扰质粒MRP2siRNA逆转人结肠癌耐药细胞株HT29/LOHP的多药耐药,为硅纳米作为靶向基因治疗和化疗药物的有效性打下基础。方法制备硅纳米载体,用硅纳米载体和脂质体分别将多药耐药基因MRP2的干扰重组质粒pGensil-MRP2siRNA,转染人结肠癌耐药细胞株HT29/L-OHP,观察两种载体转染人结肠癌耐药细胞株HT29/L-OHP后绿色荧光的表达,利用RT-PCR和Western-blot检测MRP2的表达,CCK-8实验测定转染干扰质粒后顺铂对人结肠癌耐药细胞株HT29/L-OHP细胞增殖抑制作用,并比较两种载体的逆转效率。结果在人结肠癌耐药细胞株HT29/L-OHP中,两种载体携带的siRNA都明显抑制了MRP2 mRNA及蛋白的表达,两种载体逆转效果差异无统计学意义(P>0.05)。在相同浓度化疗药物的作用下,两种载体的RNA干扰组细胞凋亡比例显著高于对照组(P<0.01),表明细胞耐药性显著下降,两种载体细胞凋亡差异无统计学意义(P>0.05)。结论硅纳米能有效的携带pGensil-MRP2siRNA质粒转染人结肠癌细胞人结肠癌耐药细胞株HT29/LOHP,并且对结肠癌耐药细胞的MRP2的表达有明显抑制作用,取得良好的逆转耐药效果。Objective To study the ability of silica nanoparticles carrying the plasmid of RNAi MRP2 in the reversal of multidrug resistance of colorectal cancer cell line,and observe the effect of the vector of MRP2 and the change of sensibility to chemotherapeutics in HT29/L-OHP cell line.Methods The recombinant plasmid was transfected into HT29/L-OHP cell by liposome and silica nanoparticle respectively and the difference of two vectors was compared.The MRP2 expression was analyzed by real time PCR and Western blot.CCK-8 test was applied to measure the inhibition combined with cis-platinum complexes proliferation of HT29/L-OHP strain.Results The over expression of MRP2 was suppressed efficiently by the vectors of liposome and silica nanoparticle respectively carrying small interfering RNA,which caused sequence specific gene silence,and the difference of two vectors was not significant （P ＞ 0.05）.The apoptosis rate of HT29/L-OHP cells was increased significantly in the siRNA group during the chemotherapy （P＜0.01）,which indicated the sensitivity to drugs was decreased significantly.Conclusions The silica nanoparticles can be used as the vector of plasmid of pGensil-MRP2siRNA and can efficiently transfect the multidrug resistance of cis-platinum complexes in colorectal cancer cell line.The RNAi can inhibit MRP2 expression effectively and reverse the multidrug resistance.

关 键 词：硅纳米 多药耐药相关蛋白基因 小RNA分子 

分 类 号：R735.35[医药卫生—肿瘤]