摘要
目的 研究反义RNA对成纤维细胞胶原蛋白的调控作用 ,探讨基因治疗增生性瘢痕的可能性。方法 将α1 (Ⅰ )型前胶原基因片段反向克隆至非整合型哺乳动物表达载体pREP9的多克隆位点中 ,以构建α1 (Ⅰ )型前胶原基因反义RNA表达载体pREP9 AScoll,并籍脂质体 lipofectamine将其导入人胚肺HLF细胞中 ,运用MTT试验、电镜、3H 脯氨酸掺入法检测反义RNA表达载体对HLF细胞的影响。结果 外源重组体pREP9 AScoll对胶原蛋白的合成有明显抑制作用 ,但对细胞增殖无任何影响。结论 α1 (Ⅰ )型前胶原基因反义重组质粒能有效调控胶原蛋白的合成 ,从而有在基因水平防治瘢痕的前景。
Objective It has demonstrated that fibroblasts from hypertrophic scar overexpressed collagen mRNA and protein , especially type Ⅰ collagen. This work was to investigate the effect of antisense RNA of α-1 (Ⅰ) collagen on human lung fibroblasts. Methods The recombinant antisense expressive plasmid of α-1 (Ⅰ) precollagen gene was constructed by inserting the partial sequence of this gene into the multiple cloning sites of vector pREP 9 inversely. Then, it was transferred into human lung fibroblasts (HLF) through lipofectamine mediation. The results were obtained from the MTT test, electron microscope and 3H|Proline incorporation. Results The data indicated that the antisense RNA of α-1 (Ⅰ) precollagen could inhibit the synthesis of α-1 (Ⅰ) collagen, but there was no significant change in their growth curves. Conclusion α-1 (Ⅰ) collagen gene expression can be inhibited by antisense nucleic acids.;
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2001年第3期157-160,共4页
Chinese Journal of Plastic Surgery
基金
广东省重点攻关项目基金!资助 (资助号 :982 78172 )
