RNA干扰抑制大鼠肌腱细胞中V型胶原基因的表达  

Inhibition of Gene Expression of Type V Collagen in Rat Tendon Cells by RNA Interference

作　　者：张国荣[1] 孙冬梅[1] 宋佰慧[1] 陈娟[1] 朱莉[1] 

机构地区：[1]长春医学高等专科学校,吉林长春130031

出　　处：《中国医学创新》2013年第28期1-3,共3页Medical Innovation of China

基　　金：吉林省教育厅"十二五"科学技术研究项目(吉教科合字[2011]第364号)

摘　　要：目的:利用RNA干扰技术(RNAi)下调肌腱细胞中V型胶原蛋白的表达,拟为后续探讨V型胶原在损伤肌腱修复过程中的作用提供可行的试验方法。方法:分别设计干扰大鼠V型胶原[COL5(1)2(2)]两个亚基COL5 1和COL5 2表达的siRNA序列,转染大鼠肌腱细胞。通过实时荧光定量PCR和免疫荧光法检测V型胶原在基因和蛋白水平的表达。结果:转染特定的siRNA后,V型胶原两个亚基的基因表达被抑制约70%,蛋白表达也被明显抑制。结论:RNAi法可有效地抑制大鼠肌腱细胞中V型胶原的基因和蛋白水平表达。为进一步研究V型胶原在肌腱损伤修复过程中的作用提供了可行的试验方法。Objective： Down regulate type V collagen expression in tendon cells using RNA interference （ RNAi ） to provide a viable test method for investigating the role of type V collagen in the injured tendon repair process. Method： Two target siRNA sequences for mRNA of the al and a2 chains in rat type V collagen were designed and transfected into rat tendon cells. Real time PCR and immunofluorescence were used to detect type V collagen expression in gene and protein level. Result： The results showed that the interference effect of the mRNA expression was about 70 % and protein level was siguificantly inhibited after specific siRNA interfection. Conclusion： RNAi method can effectively inhibit the gene and protein expression of type V collagen in rat tendon cells. This study provides a viable test method to further study the role of type V collage in injured tendon repair process.

关 键 词：肌腱细胞 V型胶原 RNA干扰技术 基因表达 

分 类 号：R346[医药卫生—基础医学]