ABSTRACT
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.
ABSTRACT
In order to explore the effects of exogenous human telomerase reverse transcriptase (hTERT/hTRT/hEST2) on telomeric restriction fragment (TRF), telomerase activity and its subunits expression in human embryonic fibroblasts (hEFs), hTERT sense eukaryotic expression vector pIRES2 EGFP hTERT was constructed with DNA recombinant technique and then transfected into primary hEFs by Lipofectin method. TRF length, telomerase activity and changes in telomerase subunits expression were examined and evaluated in transfected and untransfected cells. The results showed that telomerase activity in pIRES2 EGFP hTERT transfected cells (hEF EGFP) was significantly higher than that in untransfected hEFs and vacant vector transfected cells (hEF EGFP) ( P