Proteomic analysis of human bone marrow mesenchymal stem cells transduced with human telomerase reverse transcriptase gene during proliferation

Abstract

Objectives: Previous studies have reported immortalization and tumorigenicity of human mesenchymal stem cells (hMSCs) transduced with exogenous human telomerase reverse transcriptase (hTERT). We also have established a line of hMSCs transduced with hTERT (hTERT-hMSCs) and we have cultured these cells for 290 population doublings (PDs) during which they demonstrated a large proliferation potential but with no tumorigenicity. The aim of this study was to investigate the protein expression profile of hTERT-hMSCs with two-dimensional gel electrophoresis and peptide mass fingerprinting by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, to be able to analyse the effects of exogenous hTERT on protein expression in hMSCs. Materials and methods: We generated proteome maps of primary hMSCs and hTERT-hMSCs at PD 95 and PD 275. Results: A total of 1543 ± 145 protein spots in gels of primary MSCs at PD 12, 1611 ± 186 protein spots in gels of hTERT-hMSCs at PD 95 and 1451 ± 126 protein spots in gels of hTERT-hMSCs at 275 PD were detected. One hundred of these were successfully identified, including 20 which were differentially expressed. Conclusions: The results suggest that sustaining levels of prohibitin and p53 expression along with differential expression of proteins in hTERT-hMSCs provide an insight into lack of transforming activity of hTERT-hMSCs during cell proliferation. © 2008 The Authors.