纳米微粒介导Bcl—xl小干扰RNA诱导人肺腺癌细胞凋亡  

Effect of nanoparticles containing Bcl-xl small interference RNA on apoptosis of human lung adenocarcinoma cells

作　　者：廖洪映[1] 谷力加[1] 陈秀玲[1] 翁毅敏[1] 李昀[1] 张建[1] 蔡松旺[1] 陈惠国[1] 宋江平[1] 

机构地区：[1]中山大学附属第三医院心胸外科,广州510630

出　　处：《中华生物医学工程杂志》2008年第4期265-268,共4页Chinese Journal of Biomedical Engineering

基　　金：广东省科技厅计划项目（2008B030301315）

摘　　要：目的探讨纳米微粒介导的Bcl—xl小干扰RNA（siRNA）对人肺腺癌A549细胞凋亡的影响。方法制备包载Bcl—xl siRNA的纳米微粒，体外培养人肺腺癌A549细胞。以纳米微粒介导将人工合成的Bcl—xl siRNA转染细胞后，将细胞分为4组：生理盐水组、纳米微粒组、siRNA-c-纳米微粒组及siRNA-纳米微粒组，其中siRNA-c-纳米微粒组及siRNA-纳米微粒组又分为0．1、0．2、0．4、0．8μmol／L4个浓度组。应用TUNEL法检测肺癌细胞凋亡；RT—PCR半定量检测Bcl—xl mRNA的表达；免疫细胞化学染色法检测Bcl—xl蛋白的表达。结果转染Bcl—xl siRNA后，各处理浓度组的肺腺癌细胞凋亡指数随浓度的增加而逐渐升高[0．1、0．2、0．4、0．8μmol／L组分别为（19．7±2．6）％、（32．4±5．5）％、（46．0±5．3）％和（55．4±5．9）％]，与生理盐水组、纳米微粒组及同浓度的siRNA—e-纳米微粒组相比，差异均有统计学意义（P均〈0．05）。转染浓度为0．8μmol／L时，肺腺癌细胞内Bcl—xl mRNA及其蛋白的表达明显降低（P〈0．05），Bcl—xl蛋白表达阳性单位下降至0．0787±0．0233；Bcl—xl mRNA表达下降至0．856±0．242，与其它各组差异有统计学意义（P〈0．05）。结论纳米微粒能有效介导Bcl—xl siRNA进入肺腺癌细胞。Bcl-xl siRNA能够特异性减少肺腺癌细胞的Bcl—xl的mRNA和蛋白的表达，抑制肿瘤细胞增殖及诱导细胞凋亡。Objective To study the effect of nanoparticle-mediated Bcl-xl small interference RNA （siRNA） on apoptosis of human lung adenocarcinoma A549 cell. Methods Nanoparticles containing Bcl-xl siRNA was prepared. A549 cells were cultured in vitro. The Bcl-xl siRNA was transfected into cultured cells by nanopaticles. The cells were divided into 4 groups： saline group, nanoparticles group, siRNA-c- nanoparticles group and siRNA-nanoparticles group. The latter two groups were also divided into 0.1,0.2, 0.4,0. 81,inol/L groups. Apoptosis changes after transfection was observed by TUNEL. Bcl-xl mRNA expression was assessed by RT-PCR, and Bcl-xl protein expression was measured by immunochemistry. Results After transfection with Bcl-xl siRNA, the apoptotic index of lung cancer cells were increased with concentrations[ the apoptotic index in 0. 1, 0. 2,0.4,0. 8 p, mol/L group were （19.7± 2. 6）%, （32.4 ± 5.5） % , （ 46.0 ± 5.3 ） % and （ 55, 4 ±5.9 ） % respectively ]. Compared with saline group, nanoparticles group, siRNA-c-nanoparticles group, was significantly different （ P 〈 0.05 ）. At the concentration of 0. 8 μmol/L, the expression level of Bcl-xl mRNA and protein reduced obviously. The positive unit of Bcl-xl protein expression decreased to 0. 0787 ± 0. 0233, and the Bcl-xl mRNA to 0. 856 ± 0. 242, which were significantly different compared to other groups （ P 〈 0.05 ）. Conclutions Bcl-xl siRNA is effectively transfected into cultured cells mediated by the nanopaticles. The Bcl-xl siRNA can specially down-regulate Bcl-xl mRNA and protein expression, inhibit the proliferation of tumor cells and induce cells apoptosis.

关 键 词：RNA干扰 纳米微粒 Bcl—xl 细胞凋亡 肺肿瘤 腺癌 

分 类 号：R734.2[医药卫生—肿瘤]