Wnt5a基因慢病毒的构建与RSC96雪旺细胞的感染研究  

Study on construction of Wnt5a gene lentivirus and infection of RSC96 Schwann cells

作　　者：韦良臣[1,2] 杨琪 曾晖 翁鉴[1,2] 刘佩 康斌[1,2] 钟华戈 于斐[1,2] WEI Liangchen;YANG Qi;ZENG Hui;WENG Jian;LIU Pei;KANG Bin;ZHONG Huage;YU Fei(Department of Bone&Joint Surgery,Peking University Shenzhen Hospital,Shenzhen,Guangdong 518036,China;National&Local Joint Engineering Research Center of Orthopedic Biomaterials,Shenzhen,Guangdong 518036,China;Department of Medical Ultrasound,Peking University Shenzhen Hospital,Shenzhen,Guangdong 518036,China;Department of Gastrointestinal Surgery,Guangxi Tumor Hospital,Nanning,Guangxi 530021,China)

机构地区：[1]北京大学深圳医院骨关节科,广东深圳518036 [2]骨科生物材料国家地方联合工程研究中心,广东深圳518036 [3]北京大学深圳医院超声诊断科,广东深圳518036 [4]广西医科大学附属肿瘤医院胃肠外科,广西南宁530021

出　　处：《现代医药卫生》2022年第4期546-550,554,共6页Journal of Modern Medicine & Health

基　　金：国家自然科学基金项目(82102568,82102076,82172432,82001319);广东省科学技术厅基础与应用基础研究基金项目(2021A1515012586,2019A1515110983,2019A1515011290);白求恩·石药骨质疏松科研基金项目(G-X-2020-1107-21)。

摘　　要：目的 构建大鼠Wnt5a基因慢病毒并感染RSC96雪旺细胞,观察慢病毒感染情况及Wnt5a表达情况。方法 设计大鼠Wnt5a基因特异性RNAi序列,构建并包装成可感染细胞的慢病毒,感染RSC96雪旺细胞后通过Celigo观察细胞中荧光表达,并通过实时荧光定量聚合酶链反应(RT-q PCR)检测细胞中Wnt5am RNA的表达情况。结果 构建的Wnt5a基因慢病毒在感染RSC96雪旺细胞后可表达绿色荧光,细胞中Wnt5am RNA的平均表达量为0.206±0.056。结论 Wnt5a基因慢病毒构建成功,可为研究Wnt5a调控的周围神经损伤修复时雪旺细胞功能的相关研究提供便利。Objective To construct rat Wnt5a lentivirus and infect RSC96 Schwann cells,and to observe the infection and the expression of Wn5a lentivirus.Methods The specific RNAi sequence of rat Wnt5a gene was designed,lentivirus that could infect cells were constructed and packaged.After RSC96 Schwann cells were infected,the fluorescent expression in cells was observed by Celigo,and the expression of Wnt5a mRNA in cells was detected by Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results The constructed Wnt5a lentivirus could express green fluorescence after infecting RSC96 Schwann cells.The average expression of Wnt5a mRNA in RSC96 Schwann cells was 0.206±0.056.Conclusion Wnt5a lentivirus was successfully constructed,to facilitate the studying on the function of Schwann cells in the repairing of peripheral nerve injury regulated by Wnt5a.

关 键 词：WNT5A 慢病毒 周围神经损伤修复 RSC96雪旺细胞 细胞实验 

分 类 号：R641[医药卫生—外科学]