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| 靶向VEGF发夹状RNA对宫颈癌HeLa细胞抑制作用的研究 | |
| 引用本文： | 庞然然,邢丽娜. 靶向VEGF发夹状RNA对宫颈癌HeLa细胞抑制作用的研究[J]. 中华肿瘤防治杂志, 2011, 18(16) |
| 作者姓名： | 庞然然 邢丽娜 |
| 作者单位： | 哈尔滨医科大学附属第二医院肿瘤放疗科,黑龙江哈尔滨,150086 |
| 基金项目： | 哈尔滨市科技攻关计划项目(2005AA9CS116-10) |
| 摘 要： | 目的:运用RNAi技术下调血管内皮生长因子(VEGF)在HeLa细胞中的表达,观察其对肿瘤细胞凋亡的影响,为人宫颈癌治疗提供理论依据。方法:设计并构建针对VEGF的携带绿色荧光蛋白(GFP)发夹状RNA(shRNA)质粒表达载体(PGPU6/GFP/Neo-shRNA),脂质体法转染HeLa细胞;荧光显微镜观察GFP的表达,并计算转染效率;RT-PCR检测HeLa细胞VEGF的表达,筛选出靶序列;再用流式细胞仪法检测细胞凋亡。结果:构建的PGPU6/GFP/Neo载体成功转入HeLa细胞;转染48h后,HeLa-shVEGF1组HeLa细胞VEGFmRNA表达的抑制率为75.0%;与HeLa组和HeLa-shNC组相比,HeLa-shVEGF1组HeLa细胞凋亡率明显增加,P<0.01。结论:本研究构建的PGPU6-shRNA表达载体携带GFP便于观察细胞的转染情况,且不影响U6启动子的转录,同时有效沉默了VEGF基因,明显增加HeLa细胞的凋亡,为未来肿瘤的治疗提供新途径。 |
| 关 键 词： | 血管内皮生长因子 RNA干扰 发夹状RNA HeLa细胞 凋亡 |
Inhibitory effect of small hairpin RNA targeting at VEGF on cervical carcinoma cell line HeLa |
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| PANG Ran-ran,XING Li-na. Inhibitory effect of small hairpin RNA targeting at VEGF on cervical carcinoma cell line HeLa[J]. Chinese Journal of Cancer Prevention and Treatment, 2011, 18(16) | |
| Authors: | PANG Ran-ran XING Li-na |
| Affiliation: | PANG Ran-ran,XING Li-na Department of Radiotherapy,Second Affiliated Hospital of Harbin Medical University,Harbin 150086,P.R.China |
| Abstract: | OBJECTIVE: To study apoptosis of the tumour cells by RNA interference(RNAi) technology to depress human vascular endothelial growth factor(VEGF) gene expression in HeLa cells,and provide a theoretical basis for the treatment of human cervical carcinoma.METHODS: Short hairpin RNA targeting at VEGF gene was designed and synthesized,which was integrated into plasmid expression vector(PGPU6) with green fluorescent protein(GFP),then transfected into HeLa cells by lipofectamine.The expression of GFP and calculate... |
| Keywords: | vascular endothelial growth factor RNA interference short hairpin RNA HeLa cell apoptosis |
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