RNAi沉默Rab5a基因对乳腺癌细胞MCF-7侵袭能力的的影响  

Influence of RNA interference targeting Rab5a on invasion of breast cancer cells MCF- 7

作　　者：查尼尔[1,2] 李志高[2] 谷新悦[2] 

机构地区：[1]内蒙古自治区人民医院肿瘤外科,呼和浩特010017 [2]哈尔滨医科大学附属肿瘤医院乳腺外科

出　　处：《实用肿瘤学杂志》2010年第3期212-216,共5页Practical Oncology Journal

基　　金：黑龙江省科技厅科技攻关项目（GC07C350）

摘　　要：目的探讨miRNA表达载体靶向沉默Rab5a表达对人乳腺癌细胞MCF-7侵袭能力的影响。方法设计4条miRNA干扰载体分别转染乳腺癌细胞MCF-7。应用RT—PCR和Western blot筛选出一组抑制效率最高的miRNA干扰载体进行Transwell小室实验检测细胞侵袭能力。结果转染Rab5a-miRNA-2干扰载体对MCF-7细胞Rab5a表达抑制效果最明显，RT—PCR结果显示其对Rab5a基因mRNA抑制率达到（50．01±2．69）％，Western Blot结果显示其对Rab5a蛋白抑制率达到（55．58±3．16）％。选择Rab5a—miRNA-2组干扰载体进行后续实验。Transwell实验显示，Rab5a—miRNA-2组穿膜细胞数为67．8±12．03，与阴性对照组和空白对照组之间均有显著性差异（P〈0．01）。结论RNAi靶向沉默Rab5a基因可以抑制乳腺癌细胞MCF-7的侵袭能力。Objective To explore the influence of RNA interference targeting RabSa on invasion of breast cancer cells MCF - 7. Method Four miRNA interference vectors targeting RabSa were designed, constructed and transfeeted into breast cancer cell line MCF - 7 each other. RT - PCR and Western blot were applied to seleete the most efficient group of miRNA interference suppression ,then transwell assay were applied to analyse the cell invasiveness. Results RT - PCR analysis showed that cells MCF - 7 transfected with RabSa - miRNA - 2 vector had the strongest inhibition of RabSa gene, with the inhibition rate （50.01 ± 2.69）%. Western blot analysis showed that cells transfected with Rab5a- miRNA -2 vector had the strongest inhibition of RabSa protein ,with the inhibition rate （68.49 ± 3.70）%. Then we used RabSa-miRNA -2 group for transwell assay. Transwell assay showed that the number of cells passing the membranes was 67.8 ± 12.03 at 48h since transfecting,and was significantly different from the negative controls and blank controls（P 〈0.01 ）. Conclusion RabSa silenced by miRNA expression vector can inhibit the invasiveness of breast cancer cells MCF -7.

关 键 词：RNA干扰 miRNA表达载体 RAB5A 乳腺癌 细胞侵袭 

分 类 号：R737.9[医药卫生—肿瘤]