慢病毒介导的E2F-1沉默对人胃癌细胞MGC803中药人参黄芪复方药物敏感性的影响  

Lentivirus-mediated E2F-1 gene silencing enhances sensitivity of gastric carcinoma MGC-803 cells to compound ginseng and astragalus

作　　者：罗文[1] 吴锟[1] 王晓通[1] 谢玉波[2] 肖强[1] 

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,广西壮族自治区南宁市530021 [2]广西医科大学第一附属医院麻醉科,广西壮族自治区南宁市530021

出　　处：《世界华人消化杂志》2013年第17期1636-1641,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目,No.30860273~~

摘　　要：目的:利用重组慢病毒介导的E2F-1基因沉默载体感染人胃癌MGC-803细胞,观察其对中药人参黄芪复方药物敏感性的影响.方法:将胃癌MGC-803细胞接种于培养板中分为4组:正常组(正常培养基)、空白对照组(中药培养基)、阴性对照LV-RNAi组(中药培养基+感染空载体慢病毒颗粒2.0×109TU/mL)、E2F-1-RNAi-LV组(中药培养基+感染E2F-1基因重组慢病毒颗粒2.0×109TU/mL),分别采用半定量逆转录-聚合酶链反应(RT-PCR)和Western blot检测E2F-1mRNA及蛋白表达;MTT方法检测细胞的增殖抑制率;Giemsa染色检测细胞的克隆形成;流式细胞仪检测细胞的凋亡;细胞划痕实验检测细胞迁移能力.结果:E2F-1基因沉默蛋白及mRNA表达水平均明显下降,差异有统计学意义(0.384±0.036vs0.883±0.051和0.923±0.049,0.220±0.056vs0.729±0.021和0.712±0.037,P<0.05).中药人参黄芪复方能抑制人胃癌MGC-803细胞生长、诱导胃癌细胞凋亡、抑制细胞克隆形成及迁移;与空白对照组及阴性对照组相比较,E2F-1基因沉默组MGC-803细胞增殖抑制率、细胞凋亡率显著提高,其细胞克隆形成、细胞迁移面积显著降低,差异均有统计学意义(0.789±0.013vs0.484±0.060和0.454±0.006,0.383±0.027vs0.114±0.038和0.089±0.051,0.024±0.003vs0.036±0.004和0.052±0.002,0.553±0.038vs0.897±0.020和0.928±0.051,P<0.05).结论:重组慢病毒载体介导的E2F-1基因沉默能有效增强中药人参黄芪复方对人胃癌MGC-803细胞的药物敏感性.AIM： To observe the effect of recombinant lentivirus-mediated E2F-1 gene silencing on sensitivity of gastric carcinoma MGC-803 cells to compound ginseng and astragalus. METHODS： MGC-803 cells were plated and divided into four groups： a normal control group （medicine medium）, a blank control group, a negative LV-RNAi control group （medicine medium ＋ infection with empty lentiviral vector particles 2.0 × 10 9 TU/mL） and an E2F-1-RNAi-LV group （medicine medium ＋ infection with recombinant E2F-1 lentiviral particles 2.0 × 10 9 TU/mL）. The mRNA and protein expression levels of E2F1 were detected by RT-PCR and Western blot. Cell proliferation was detected by MTT assay. Cell colony formation was detected by Giemsa staining. Cell apoptosis was observed by flow cytometry, and cell migration was detected by cell scratch assay. RESULTS： Infection with the recombinant lentiviral vector significantly decreased the protein and mRNA expression levels of E2F-1 compared to the two control groups （0.384 ± 0.036 vs 0.883 ± 0.051 and 0.923 ± 0.049, 0.220 ± 0.056 vs 0.729 ± 0.021 and 0.712 ± 0.037, P 0.05）. Compound ginseng and astragalus inhibited MGC-803 cell growth, induced cell apoptosis, and reduced cell colony formation and migration. Compared to the two control groups, the rates of reduced cell colony formation and cell apoptosis were significantly increased and the areas of cell colony formation and migration were significantly decreased in the E2F-1 gene silencing group （0.789 ± 0.013 vs 0.484 ± 0.060 and 0.454 ± 0.006, 0.383 ± 0.027 vs 0.114 ± 0.038 and 0.089 ± 0.051, 0.024 ± 0.003 vs 0.036 ± 0.004 and 0.052 ± 0.002, 0.553 ± 0.038 vs 0.897 ± 0.020 and 0.928 ± 0.051, P 0.05）. CONCLUSION： Recombinant lentivirus-mediated E2F-1 gene silencing effectively enhances the sensitivity of MGC-803 cells to compound ginseng and astragalus.

关 键 词：中药人参黄芪复方 E2F-1 人胃癌MGC803细胞 药物敏感性 

分 类 号：R735.2[医药卫生—肿瘤]