Survivin、TGFβ3与TIMP-1慢病毒载体的构建及其人髄核细胞表达  

CONSTRUCTION OF SURVIVIN,TGFβ3 AND TIMP-1 LENTIVIRAL VECTOR AND DETECTION OF THEIR EXPRESSIONS IN HUMAN NUCLEUS PULPOSUS

作　　者：郭柱[1] 岳斌[1] 马学晓[1] 胡有谷[1] 杨堃[1] 陈伯华[1] 

机构地区：[1]青岛大学医学院附属医院脊柱外科,山东青岛266003

出　　处：《青岛大学医学院学报》2014年第5期380-382,共3页Acta Academiae Medicinae Qingdao Universitatis

基　　金：国家自然基金资助项目(81171758)

摘　　要：目的构建人生存素（Survivin）、转化生长因子03（TGFtβ3）与基质金属蛋白酶抑制剂-1（TIMP-1）三基因过表达慢病毒载体，通过一次转染，实现Survivin、TGFβ3、TIMP-1三基因在人退变椎间盘髓核细胞的稳定转染，为逆转或延缓椎间盘退变的基因治疗提供实验基础。方法应用Survivin、TGFβ3、TIMP-1慢病毒载体联合转染人退变髓核细胞，应用Real—TimePCR技术检测转染后基因表达情况。结果人退变椎间盘髓核细胞在转染后1周，Survivin、TGFβ3、TIMP-1三基因在基因水平均获得稳定过表达，并且其表达能维持细胞传至第3代或更久。Real—TimePCR检测结果显示，目的病毒组Survivin、TGFβ3、TIMP-1均过表达，其相对表达量与空白组比较，差异有显著性（F=12．96～3889．70，P〈O．05）。结论Survivin、TGFβ3与TIMP-1慢病毒载体能够稳定转染人退变椎间盘髓核细胞，并且能长时间维持其表达，从而发挥生物学效应。Objective To construct Survivin,TGFβ3 and TIMP-1 three-gene lentiviral vector and to achieve stable trans fection of the three genes in human degenerative intervertebral disc nucleus pulposus through one single transfection, providing experimental basis for gene therapy of reversing or slowing down the degeneration of intervertebral disc. Methods Transfecting human degenerative intervertebral disc nucleus pulposus by using Survivin, TGF〈3 and TIMP-1 lentiviral vector, the expressions of the three genes were detected by real-time PCR technique. Results Survivin,TGFβ3 and TIMP-1 obtained stable over expres sion after one week of transfection and their expressions could last to third generation or longer. Real-time PCR indicated that the three gens were all over expressed, the differences were significant as compared with the normal control group with regard to their relative expression （F=12.96—3 889.70,P〈0.05）. Conclusion Survivin, TGFβ3 and TIMP-1 three-gene lentiviral vector can stably transfect the nucleus pulposus cells of degenerative intervetebral disk and last for a long time to maintain the expressions, and thus develop their biological effect.

关 键 词：凋亡抑制蛋白质类 转化生长因子Β3 金属蛋白酶1组织抑制剂 慢病毒属 椎间盘 退变 

分 类 号：R681.53[医药卫生—骨科学]