人端粒酶催化亚单位基因转染人胚胎成纤维细胞后胶原的变化

The Effects of hTERT Transfection on the Collagen Expression of Human Embryonic Fibroblasts

目的 将重组正义hTERT基因荧光真核表达载体导入人胚胎成纤维细胞 (hEF)中 ,探讨外源性hTERT基因转染对hEF细胞胶原含量变化的影响。方法 应用脂质体法将正义重组pIRES2 -EGFP -hTERT质粒及空载pIRES2 -EGFP质粒分别转染至原代培养hEF中 ,westernblot检测转染细胞和未转染细胞中Ⅰ、Ⅲ型胶原表达变化 ;放免法检测转染细胞和未转染细胞培养液上清中Ⅰ、Ⅲ型胶原含量。结果 外源性hTERT基因转染细胞 (hEF -hTERT)较未转染细胞 (hEF)和空载体转染细胞 (hEF -EGFP)中Ⅰ、Ⅲ型胶原表达明显增加 ;hEF -hTERT细胞培养上清液中Ⅰ、Ⅲ型胶原含量也较hEF和hEF -EGFP细胞有显著增加。结论 外源性hTERT基因转染能使原代培养的hEF细胞胶原合成增加 。

Objective The recombinant fluorescent eukaryotic expressing vector containing hTERT cDNA was transfected into human embryonic fibroblasts (hEFs) to explore the effects of exogenous hTERT on the type I and III collagens expression in hEFs. Methods p IRES2-EGFP-hTERT plasmid and pIRES2-EGFP plasmids were transfected into primary hEFs respectively by Lipofectin reagent. Expression of type I and III collagen was determined by Western blotting and the content of type I and III collagens in the cellular medium at 3 days after transfection were examined by radio-immunoassay. Results The expression levels of type I and III collagens in hTERT gene transfected hEFs(hEF-hTERT) were obviously higher than those in untransfected hEFs and vacant vector transfected hEFs(hEF-EGFP). The content of type I and III collagens in the cellular medium in hEF-hTERT cells at 3 days after transfection was also higher than that in untransfected hEFs and hEF-EGFP cells. Conclusions The synthesis ability of type I and III collagens in hEFs could be promoted by exogenous hTERT gene transfection.