| 实时荧光定量PCR 检测人肿瘤细胞NKG2D 配体基因表达 |
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| 引用本文： | 刘枫,郑冰蓉,杨举伦,王力,陈玥,赵稳兴.实时荧光定量PCR 检测人肿瘤细胞NKG2D 配体基因表达[J].现代生物医学进展,2011,11(19):3621-3624. |
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| 作者姓名： | 刘枫 郑冰蓉 杨举伦 王力 陈玥 赵稳兴 |
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| 作者单位： | 1. 云南大学生命科学学院 云南昆明650091
2. 解放军昆明总医院病理科 云南昆明650032 |
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| 摘 要： | 目的:建立人肿瘤细胞NKG2D配体基因(MICA、MICB、ULBP1、ULBP2、ULBP3)表达的实时荧光定量PCR(real-timefluorescence quantitative PCR)检测方法。方法:根据NCBI基因库中NKG2D配体基因序列,设计合成引物。用Trizol法从培养的肿瘤细胞(BEC-7402、HeLa、MDA-MB-435、XWLC-05)中提取总RNA,逆转录成cDNA,建立实时荧光定量PCR检测NKG2D配体基因表达的方法,并检测NKG2D配体在肿瘤细胞株中的表达。结果:经过琼脂糖凝胶电泳、熔解曲线和标准曲线分析,用所设计的引物和SYBR Green I能够特异扩增和定量检测NKG2D配体基因的表达。该方法成功检测4种肿瘤细胞NKG2D配体基因的表达。结论:建立了人NKG2D配体基因表达的实时荧光定量PCR检测方法,为进一步研究人NKG2D配体在肿瘤免疫中的作用提供了有效手段。
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| 关 键 词： | NKG2D配体 实时荧光定量PCR 肿瘤细胞 |
Real-Time Fluorescence Quantitative PCR Assay for Human NKG2D Ligand
mRNA Quantification |
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| LIU Feng,ZHENG Bing-rong,YANG Ju-lun,WANG Li,CHEN Yue,ZHAO Wen-xing.Real-Time Fluorescence Quantitative PCR Assay for Human NKG2D Ligand
mRNA Quantification[J].Progress in Modern Biomedicine,2011,11(19):3621-3624. |
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| Authors: | LIU Feng ZHENG Bing-rong YANG Ju-lun WANG Li CHEN Yue ZHAO Wen-xing |
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| Institution: | LIU Feng1,ZHENG Bing-rong1,YANG Ju-lun2,WANG Li2,CHEN Yue2,ZHAO Wen-xing2(1 College of Life Science,Yunnan University,650091,Kunming,China,2 Department of pathology,Kunming general hospital of PLA,650032,China) |
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| Abstract: | Objective: This study aimed to develop a real-time fluorescence quantitative PCR assay for NKG2D ligand mRNA in human.Methods: The primers for NKG2D ligands and keeping-home gene GAPDH were designed and synthesized according to the gene sequences from NCBI,The total RNA of tumor cells was extracted by Trizol protocol,and cDNA was synthesized by reverse transcrip-tase,then the real-time fluorescence quantitative PCR assay was established using SYBR Green I.Results: The method was efficient and sensitive for ... |
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| Keywords: | NKG2D ligand Real-time fluorescence quantitative PCR Tumor cell |
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