| siRNA表达载体对肝素酶基因的沉默作用 |
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| 引用本文： | 刘启志,王烈. siRNA表达载体对肝素酶基因的沉默作用[J]. 中国普外基础与临床杂志, 2010, 17(2): 142-146 |
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| 作者姓名： | 刘启志 王烈 |
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| 作者单位： | 1. 福建医科大学福总临床医学院,福州,350025
2. 南京军区福州总医院普通外科研究所,福州,350025 |
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| 基金项目： | 南京军区"十一五"科研规划重点课题 |
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| 摘 要： | 目的根据基因库中的人肝素酶(HPSE)基因序列,构建针对肝素酶基因的小干扰RNA(siRNA)及其表达载体,转染MDA-MB-231细胞后观察其对肝素酶基因的干扰作用。方法设计肝素酶靶向的发夹状siRNA,合成2条互补的寡核苷酸链共4对,退火后连接入pGPU6/GFP/Neo载体,转化后进行序列鉴定,转染MDA-MB-231细胞,荧光照片,RT-PCR(real-time PCR)检测肝素酶mRNA的表达情况。结果设计出4条针对肝素酶的siRNA,将退火后的双链寡核苷酸片段克隆到pGPU6/GFP/Neo载体,经过阳性菌落酶切鉴定与测序,结果正确。再将siRNA载体转染到MDA-MB-231细胞株,RT-PCR检测肝素酶mRNA表达明显降低(P0.05)。结论成功构建了针对肝素酶基因的siRNA载体,转染到MDA-MB-231细胞株后细胞肝素酶基因的表达明显降低,从而为肿瘤治疗提供了一种新的方法。
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| 关 键 词： | RNA干扰 肝素酶 质粒 MDA-MB-231细胞株 转染 实时定量PCR |
Silencing of Heparanase Gene-Targeted by Small Interfering RNA |
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| LIU Qi-zhi,WANG Lie. Silencing of Heparanase Gene-Targeted by Small Interfering RNA[J]. Chinese Journal of Bases and Clinics In General Surgery, 2010, 17(2): 142-146 |
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| Authors: | LIU Qi-zhi WANG Lie |
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| Affiliation: | LIU Qi-zhi1,WANG Lie2. 1.Fuzong Medical College of Fujian Medical University,Fuzhou 350025,China,2.Research Institute of General Surgery,Fuzhou General Hospital of Nanjing Military Region |
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| Abstract: | Objective According to heparanase's gene sequence of GenBank,to construct heparanase gene-targeted small interfering RNA (siRNA) and its expression vector and to observe its interference effect on the expression of heparanase gene in human malignant breast cancer MDA-MB-231 cell. Methods Heparanase gene-targeted hairpin siRNA was designed,two complementary oligonucleotide strands were synthesized and inserted into pGPU6/GFP/Neo vector,which was identified by sequence identify. Human malignant breast cancer ... |
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| Keywords: | RNA interference Heparanase Plasmid MDA-MB-231 cell line Transfection Real-time polymerase chain reaction |
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