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Adenoviral HTERT Transfection Results in Altered Phospho‐erk Activity in Human Dermal Fibroblasts
Author(s) -
Lu Leonard,
Rosenberg David S.,
Mogford Jon E.,
Mustoe Thomas A.
Publication year - 2004
Publication title -
wound repair and regeneration
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.847
H-Index - 109
eISSN - 1524-475X
pISSN - 1067-1927
DOI - 10.1111/j.1067-1927.2004.0abstractar.x
Subject(s) - telomerase reverse transcriptase , telomerase , telomere , transfection , mapk/erk pathway , microbiology and biotechnology , biology , cell growth , phosphorylation , chemistry , cancer research , cell culture , dna , gene , biochemistry , genetics
: Telomeres are nucleoprotein structures at the ends of each chromosome. Due to the inability of DNA polymerase to replicate the full length of the chromosome, up to 50–200 base pairs of the telomere are lost during each successive round of cell division. In adult human somatic cells, telomerase is not active resulting in progressive loss of telomere length and entry into replicative senescence as observed in cell culture. hTERT is the catalytic subunit of telomerase, an enzyme which maintains telomere length. Transfection of human dermal fibroblasts (HDFs) by hTERT has been shown to reverse the senescent phenotype seen in aging HDFs in vitro . ERK (p44/42) is a MAP kinase which functions as a critical intermediary in the determination of cell growth and differentiation. Activation of ERK occurs through phosphorylation of threonine and tyrosine residues.
Methods : In order to delineate some of the cellular mechanisms by which hTERT functions, we treated adenoviral hTERT (Ad‐hTERT) transfected HDFs with TGFB1, and assayed phosphorylated ERK activity by Western blotting.
Results : Ad‐hTERT treated HDFs demonstrated a 2–3 fold increase in phospho‐ERK activity. In addition, our preliminary findings show that Ad‐hTERT transfected HDFs have increased TGFB1, TGFB1‐Receptor I and II, and COL1A1 gene expression by real‐time rtPCR.
Conclusions : Increased phosphor‐ERK activity as well as increased TGFB1, TGFB1‐Receptor I and II, and COL1A1 gene expression is seen in hTERT transfected HDF’s. Further studies will focus on defining other intermediary changes resulting from Ad‐hTERT tranfection. Funding source: Geron Corporation