三阴性乳腺癌IL-17基因Real-time RT-qPCR检测法的建立与应用  

Establishment of Real-time quantitative RT-qPCR assays for detecting IL-17 of three negative breast cancer

作　　者：孟庆杰[1] 巫姜[1] 石文龙[1] 崔风强[1] 樊菁[1] 李南林[1] 王廷[1] 凌瑞[1] 

机构地区：[1]第四军医大学西京医院甲乳血管外科,陕西西安710032

出　　处：《现代肿瘤医学》2017年第15期2357-2361,共5页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:81472598);陕西省社会发展科技公关项目(编号:2016SF-298)

摘　　要：目的:建立Real-time RT-qPCR(实时荧光定量RT-PCR)检测IL-17的方法,应用该方法检测三阴性乳腺癌(three negative breast cancer,TNBC)组织IL-17 mRNA的水平。方法:以TNBC组织为实验组,纤维腺瘤旁正常乳腺组织为对照组,并经HE染色病理分析确诊,Trizol法提取总RNA并反转录为c DNA。选β-actin作为内参,建立SYBR GreenⅠReal-time RT-qPCR检测法。利用该方法检测两组IL-17和β-actin的初始模板量,IL-17/β-actin计算IL-17 mRNA的相对表达量。结果:IL-17扩增效率为98.6%,相关系数0.997,溶解曲线为特异单峰,变异系数小于2.0%,IL-17 mRNA在TNBC组[(0.64±0.12)×10^(-2)]的相对表达高于正常对照组[(0.43±0.07)×10^(-2)],差异有统计学意义(P=0.025)。结论:成功建立了人源IL-17的Real-time RT-qPCR检测方法,TNBC组IL-17的高表达提示其可能与TNBC有一定关系,为研究TNBC的发病机制奠定了理论基础。Objective： To establish a Real-time quantitative RT-qPCR detection method of IL-17 and detect the IL-17 mRNA level of human three negative breast cancer（ TNBC）. Methods： TNBC was the experimental group while the fibroadenoma adjacent normal breast was control group. Both groups were stained by HE in order to confirme. Total RNA was extracted by Trizol and reverse transcribed into c DNA. β-actin was chose as an internal control. Establish SYBR Green I Real-time RT qPCR method and detect the initial template amount of two groups of IL-17. The relative expression of mRNA IL-17 was calculated by IL-17/β-actin. Results： IL-17 amplification efficiency was 98. 6%,correlation coefficient was 0. 997. Melting curve was specific unimodal. Coefficient of variation was less than 2. 0%. IL-17 mRNA expressed in each group as follow： TNBC group [（ 0. 64 ± 0. 12） × 10^（-2）] was higher than normal control group [（ 0. 43 ± 0. 07） × 10^（-2）]. The difference was statistically significant（ P = 0. 025）.Conclusion： The RT-qPCR Real-time detection method was successfully established. The high expression of IL-17 in the TNBC group suggested that it may be related to the TNBC,which laid a theoretical foundation for the study of the pathogenesis of TNBC.

关 键 词：三阴性乳腺癌 IL-17 实时荧光定量RT-PCR SYBR Green I 

分 类 号：R737.9[医药卫生—肿瘤]