siRNA干扰对人软骨细胞Skp2表达的影响  

Effect of small interference RNA on expression of the Skp2 in human chondrocytes cell

作　　者：袁长深[1] 李彦宏 刘晋邑 廖书宁 梅其杰[1] 徐文飞 段戡[1] YUAN Chang-shen;LI Yan-hong;LIU Jin-yi;LIAO Shu-ning;MEI Qi-jie;XU Wen-fei;DUAN Kan(Orthopedic Department of the Limbs,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Guangxi University of Chinese Medicine,Nanning 530000,China)

机构地区：[1]广西中医药大学第一附属医院四肢骨伤科,广西南宁530023 [2]广西中医药大学研究生院,广西南宁530000

出　　处：《海南医学院学报》2023年第13期983-987,996,共6页Journal of Hainan Medical University

基　　金：国家自然科学基金(82060875);国家自然科学基金(82160912);广西自然科学基金(2022JJA141229)。

摘　　要：目的:研究表达载体介导的siRNA对Skp2在人软骨细胞中表达的抑制效果。方法:利用在线siRNA设计工具设计3条Skp2序列siRNA-59、siRNA-318、siRNA-504作为靶位点,采用基因重组技术体外成功构建Skp2 siRNA表达载体,检测重组质粒转染对Skp2 mRNA表达的影响,并采用DNA电泳对其结果进行验证。结果:测序表明Skp2干扰序列正确,siRNA-59、siRNA-318、siRNA-504转染组的Skp2 mRNA表达明显低于空载组和NC组(P<0.05),Skp2 mRNA在siRNA-59、siRNA-318、siRNA-504中的抑制率分别为60%、41%、64%,且以siRNA-504转染组抑制率最高。结论:成功构建Skp2干扰真核表达载体,siRNA重组体能有效抑制人软骨细胞Skp2 mRNA表达,可为骨关节炎治疗提供有力证据。Objective:To study the inhibitory effect of siRNA mediated by expression vectors on Skp2 expression in human chondrocytes.Method:Three Skp2 sequences siRNA-59,siRNA-318 and siRNA-504 were designed as target sites using online siRNA design tools.Skp2 siRNA expression vectors were successfully constructed in vitro by gene recombination technology,and the influence of recombinant plasmid transfection on Skp2 mRNA expression was detected.DNA electrophoresis was used to verify the results.Results:The sequence of Skp2 interference was correct by sequence analysis.The expression of Skp2 mRNA in siRNA-59,siRNA-318,siRNA-504 transfection group was significantly lower than that in no-load group and NC group(P<0.05),the inhibition rates of Skp2 mRNA in siRNA-59,siRNA-318 and siRNA-504 were respectively 60%,41% and 64%,and the siRNA-504 transfection group had the highest inhibition rate.Conclusion:The siRNA eukaryotic expression vector of Skp2 gene was constructed successfully which effectively inhibit Skp2 mRNA expression in human chondrocytes cell,and can provide strong evidence for the treatment of osteoarthritis.

关 键 词：S期激酶相关蛋白2 骨关节炎 小干扰RNA 基因表达 

分 类 号：R684.3[医药卫生—骨科学]