南京医科大学学报（自然科学版）2012，Vol.32Issue(5)：626-630,5.
小鼠11β-HSD1基因过表达的前成骨细胞系的建立
Establishment of mouse 11β-HSD1 gene overexpression model in a MC3T3-E1 preosteoblast cell lines
摘要
Abstract
Objective:To establish a mouse preosteoblast cell lines overexpressing l1β-HSDl gene. Methods; A lentiviral vector overexpressing mouse llβ-HSDl was constructed. Lentivirus particles were generated in 293FT cells,followed by concentration and purification. MC3T3-E1 preosteoblast cells were infected by the lentivirus,which were later selected by BSD. The mRNA and protein expression of 11β-HSD1 was analyzed by real-time PCR and Western blot,respectively. MC3T3-E1 osleogenic differentiation was detected using Alizarin red staining. Results;The lentiviral vector of mouse llβ-HSDl overexpression was successfully constructed,and the MC3T3-E1 preosteoblast cell lines were infected efficiently by the lentivirus. The mRNA and protein levels of 11β-HSD1 were higher in MC3T3-E1 cells infected by lβ-HSDl overexpression lentivirus than that of cells infected by the negative control lentivirus. Alizarin red staining showed that the differentiation ability toward osteoblasts of MC3T3-EI cells was not influenced by lentivirus infection. Conclusion: Our findings showed that 11β-HSD1 gene was effectively overexpressed in MC3T3-E1 preosteoblast cells using lentivirus infection,which provides a useful tool for further studies of 11P-HSD1 function in oateoblasls.关键词
慢病毒载体/11β-HSD1/MC3T3-E1前成骨细胞系Key words
lenlivirus vector/ l1β-HSDl1 MC3T3-E1 preosteoblasl cell lines分类
生物科学引用本文复制引用
祁寒梅,吴琳,王龙,毕建华,丁国宪..小鼠11β-HSD1基因过表达的前成骨细胞系的建立[J].南京医科大学学报（自然科学版）,2012,32(5):626-630,5.基金项目
国家自然科学基金青年基金项目(30900505) （30900505）
