CXCR4-shRNA纳米复合微粒对肾癌细胞增殖的抑制作用  

Inhibitory effect of polyamidoamine dendrimer-CXCR4-shRNA nanoparticles on the proliferation of renal carcinoma cells

作　　者：董勤[1] 叶春林[1] 高烽[1] 崔杰[1] 陈培 王少良[1] 蔡晨[2] 

机构地区：[1]武警上海总队医院肾内科,上海201103 [2]第二军医大学长海医院特需诊疗科,上海200433

出　　处：《中国肿瘤生物治疗杂志》2015年第1期57-61,共5页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81202019);上海市卫生局青年科研资助项目(No.2011Y197)~~

摘　　要：目的:制备聚酰胺胺型树枝状分子(polyamidoamine dendrimer,PAMAM-D)载送CXCR4-shRNA的纳米复合微粒(PAMAM-shRNA),研究其在体外抑制人肾癌A498细胞增殖的效应。方法:将第7代的PAMAM-D室温下与CXCR4-shRNA混合(质量比为1∶0.73),制备成纳米树形分子与CXCR4-shRNA的纳米复合物PAMAM-shRNA,采用透射电镜观察PAMAMshRNA的形态结构,激光粒径仪测定其粒径。分别以PAMAM-shRNA、CXCR4-shRNA和PAMAM-D转染A498细胞,MTT法检测PAMAM-shRNA对肾癌细胞A498增殖的抑制作用,流式细胞术检测PAMAM-shRNA诱导A498细胞的凋亡情况,Real-time PCR分析转染后肾癌A498细胞CXCR4 mRNA的表达水平。结果:成功制备的新型纳米复合微粒PAMAM-shRNA分散性好,不粘连,其平均粒径为(176.5±25.48)nm。PAMAM-shRNA可以有效地抑制人肾癌细胞A498的增殖,且随着PAMAM-shRNA浓度和药物作用时间的增加,细胞增殖抑制的效果越明显,其最高增殖抑制率达(66.5±2.7)%;其还可加强诱导肾癌A498细胞凋亡。Real-time PCR检测结果表明,与CXCR4-shRNA组相比,PAMAM-shRNA组的CXCR4 mRNA的表达水平明显下降[(0.29±0.035)vs(0.70±0.084),P<0.05]。结论:PAMAM-D能高效递送CXCR4-shRNA进入A498细胞,PAMAM-shRNA以剂量和时间依赖方式显著抑制肾癌细胞增殖和诱导肾癌细胞凋亡,其在肿瘤基因治疗中具有潜在的应用价值。Objective: To optimize the preparation of polyamidoamine dendrimer-CXCR4-shRNA nanoparticles( PAMAM-shRNA) and study the inhibitory effect of PAMAM-shRNA nanoparticles on the proliferation of renal carcinoma cells in vitro. Methods: The seventh generation of polyamidoamine dendrimer( PAMAM-D) and CXCR4-shRNA were mixed at a mass ratio of 1∶ 0. 73) at room temperature. The morphology and structure of PAMAM-shRNA by transmission electron microscopy,and determined the size of the nanoparticles were analyzed by laser particle size analyzer. Human renal carcinoma A498 cells were transfected with PAMAM-shRNA,CXCR4-shRNA and PAMAM-D respectively. After transfection,cell viability was assessed by MTT assays,cell apoptosis by flow cytometry,and CXCR4 mRNA abundance by Real-time PCR. Results: The prepared PAMAM-shRNA nanoparticles were evenly distributed and non-adherent with a mean diameter of 176. 5 ± 25. 48 nm. PAMAM-shRNA effectively inhibited A498 proliferation in time- and dose-dependent manners,and the highest proliferation inhibition rate was up to( 66. 5 ± 2. 7) %. PAMAM-shRNA induced A498 cell apoptosis.CXCR4 mRNA abundance was significantly decreased( P < 0. 05) in cells transfected with PAMAM-shRNA( 0. 29 ±0. 035) than in cells transfected with CXCR4-shRNA( 0. 70 ± 0. 084). Conclusions: PAMAM dendrimers may efficiently mediate the entry of CXCR4-shRNA into renal carcinoma cells,where they exert proliferation-inhibitory and apoptosis-promoting activities. These observations suggest that PAMAM-shRNA nanoparticles may have a potential clinical application in gene therapy of renal cancer.

关 键 词：聚酰胺胺型树枝状分子 CXCR4 短发夹RNA 肾癌 

分 类 号：R737.11[医药卫生—肿瘤]