[Influence of human telomerase reverse transcriptase gene transfection on the proliferation of human embryonic fibroblasts].
OBJECTIVE
To investigate the influence of human telomerase reverse transcriptase (hTERT) gene transfection on the proliferation of human embryonic fibroblasts (hEF).


METHODS
hEFs were cultured in vitro. Sense recombinant eukaryotic plasmid (pIRES2-EGFP-hTERT) and pIRES2-EGFP vacant vector were transfected into hEF respectively with Lipofectin reagent, and were named as hEF-hTERT and hEF-EGFP. The hTERT, Id1, PCNA and I, III type collagen expression in these cells were detected by Western blot. Then the cell cycle and growth curve were measured and plotted with flow cytometry and MTT method, respectively.


RESULTS
1. The expression of hTERT, Id1, PCNA, type I and III collagen in hEF-hTERT were much higher than that in hEF and hEF-EGFP. 2. As shown in the growth curve, the OD value of hEF-hTERT at 4 to 6 days after culture was obviously higher than that of hEF and hEF-EGFP (P < 0.05), while no difference existed between hEF and hEF-EGFP from 1 to 6 days after culture (P > 0.05). 3. The cell number in G0/G1 phase in hEF-hTERT was less than that in hEF and hEF-EGFP. The cell number of hEF-hTERT in S and G2/M phase and its proliferation index (57.47%) increased when compared with that in hEF-EGFP (13.13%) and hEF (17.38%), but there was no difference between hEF and hEF-EGFP.


CONCLUSIONS
Exogenous hTERT gene transfection could promote the proliferative capacity of hEF.
Liang
Guang-Ping
GP
Institute of Burn Research, Southwest Hospital, State key Laboratory of Trauma, Burns and Combined Injury, The Third Military Medical University, Chongqing 400038. P.R. China.
Luo
Xiang-Dong
XD
Yang
Zong-Cheng
ZC
D049109:Cell Proliferation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D019098:Telomerase / Q000235:genetics*; D014162:Transfection
C509186:TERT protein, human; D019098:Telomerase