重组质粒pcDNA3/sIL-1R(Ⅰ)转染牙龈成纤维细胞的瞬时表达检测

Transient Expression of Constructed Eukaryotic Vectors Carrying Encoding Gene of Soluble Human Interleukin-1 Receptor(Ⅰ) in Human Gingival Fibroblast Cells.

目的 检测以sIL 1R胞外成熟肽编码区基因作为目的基因构建的重组质粒pcDNA3/sIL 1R(Ⅰ )对人牙龈成纤维细胞的转染效率及瞬时表达量 ,为pcDNA3/sIL 1R(Ⅰ )导入动物牙龈组织提供依据。方法 人牙龈成纤维细胞体外原代培养并传代 ,重组质粒用脂质体包裹介导进行体外转染。ELISA法对重组质粒在人牙龈成纤维细胞的表达产物进行含量检测。结果 sIL 1R(Ⅰ )在基因转染 2 4h内开始表达 ,72h最高 ,转染组细胞培养液和细胞冻融液中sIL 1R表达量均显著高于对照组 (P <0 .0 5 ) ,1周后表达逐渐减弱 ,2周后仍有少量表达。结论 通过脂质体介导的基因转染方法 ,重组质粒pcDNA3/sIL 1R(Ⅰ )在人牙龈成纤维细胞系中具有表达功能 。

Objective To detect the expression of the constructed pcDNA3 carrying encoding gene of sIL-1R(Ⅰ) transiently in human gingival fibroblast cells in vitro. Methods Human gingival fibroblast cells were transfected with recombinant plasmid pcDNA3/sIL-1R(Ⅰ) by means of liposome media methods. The protein expression products were detected by ELISA. Results The expression of sIL-1R(Ⅰ) began in 24 hours after being transferred, reached maximum in 72 hours and the protein expression products could be detected in the cell plasma and the cell culture supernatant. The level of expression in experimental groups were much higher than that in uhe control groups (P<0.05), then decreased in 1 week, but there was still a weak expression for more than 2 weeks. Conclusion The constructed pcDNA3/sIL-1R(Ⅰ) can express interest protein in human gingival fibroblast cells transiently and this establishes the basis for future investigation about the expression of pcDNA3/sIL-1R(Ⅰ) in animal's gingival tissue.