靶向沉默先天性心脏病致病基因NKX2-5  

Targets silencing congenital heart disease gene NKX2-5

作　　者：张帆 宗渊 厍广岩 钟浩[2] 武建英[2] Zhang Fan;Zong Yuan;She Guang-yan;Zhong Hao;Wu Jian-ying(Graduate School of Qinghai University,Xining 810016,China;Department of Cardiovascular Surgery,Affiliated Hospital of Qinghai University,Xining 810000,China)

机构地区：[1]青海大学研究生院,青海西宁810016 [2]青海大学附属医院心血管外科,青海西宁810000

出　　处：《兰州大学学报（医学版）》2022年第2期4-8,共5页Journal of Lanzhou University（Medical Sciences）

基　　金：国家自然科学基金面上资助项目(31571231);青海省临床医学研究中心项目(2019-SF-L2)。

摘　　要：目的 了解NKX2-5基因在细胞中的沉默表达模式。方法 构建NKX2-5基因沉默体外表达载体,将载体转染人脐静脉内皮细胞(HUVEC株),利用Western blotting和定量聚合酶链反应技术分析NKX2-5体外表达载体在细胞中的转录水平。结果 慢病毒载体在5μg/mL Polybrene条件下阳性细胞比例基本达到高峰,转染效率可达80%以上。Western blotting结果显示实验组PLVE2641表达量低于对照组PLVT7。定量聚合酶链反应结果显示实验组PLVE2641靶基因NKX2-5受到沉默,mRNA表达量显著下降,抑制率> 72%,差异有统计学意义(P<0.05)。结论 成功构建NKX2-5基因体外沉默模型,为进一步研究先天性心脏病形成奠定实验基础,为基因治疗提供理论依据。Objective To understand the silent expression pattern of NKX2-5 gene in cells. Methods Constructing NKX2-5 gene silencing in vitro expression vector, and transfecting the vector into human umbilical vein endothelial cells(HUVEC). Analyzing the transcription level of NKX2-5 in vitro expression vector by Western blotting and quantitative PCR techniques. Results The proportion of positive cells in the lentiviral vector reached a peak under the condition of 5 μg/mL Polybrene, the transfection efficiency could reach over80%. Western blotting results showed that the expression of PLVE2641 in the experimental group was lower than PLVT7 in the control group. The quantitative PCR results showed that the PLVE2641 target gene NKX2-5 in the experimental group was silenced, the mRNA expression was significantly decreased, and the inhibition rate > 72%, which was statistically significant(P < 0.05). Conclusion The successful construction of NKX2-5 gene silencing model in vitro will lay an experimental foundation for further research into the formation of congenital heart disease and provide a theoretical basis for gene therapy.

关 键 词：RNA干扰 基因沉默 先天性心脏病 

分 类 号：R446.1[医药卫生—诊断学]