Enhanced control of proliferation in telomerized cells
Tóm tắt
Clones of telomerized fibroblasts of adult human skin have earlier been obtained. It was shown that despite their fast growth in mass cultures, these cells poorly form colonies. Conditioned medium, antioxidants, and reduced partial oxygen pressure enhanced their colony formation, but not to the level characteristic of the initial cells. The conditioned medium of telomerized cells enhanced colony formation to a much greater extent than that of the initial cells. A study of proteome of the telomerized fibroblasts has revealed changes in the activities of tens of genes. A general trend consists in weakening and increased lability of the cytoskeleton and in activation of the mechanisms controlling protein degradation. However, these changes are not very pronounced. During the formation of immortal telomerized cells, selection takes place, which appears to determine changes in the expression of some genes. It was proposed that a decrease in the capacity of telomerized cells for colony formation is due to increased requirements of these cells to cell-cell contacts. The rate of cell growth reached that characteristic of mass cultures only in the largest colonies. In this respect, the telomerized fibroblasts resembled stem cells: they are capable of self-maintenance, but “escape” to differentiation in the absence of the corresponding microenvironment (niche), which is represented by other fibroblasts. Nondividing cells in the test of colony formation should be regarded as differentiated cells, since they have no features of degradation, preserve their viability, actively move, grow, phagocytize debris, etc. It was also shown that telomerization did not prevent differentiation of myoblasts and human neural stem cells. Thus, the results obtained suggest the existence of normal mechanisms underlying the regulation of proliferation in the telomerized cells, which opens possibilities of their use in cell therapy, especially in the case of autotransplantation to senior people, when the cell proliferative potential is markedly reduced and accessibility of stem cells is significantly restricted.
Tài liệu tham khảo
DuBridge, R.B., Tang, P., Hsia, H.C., et al., Analysis of Mutation in Human Cells by Using an Epstein-Barr Virus Shuttle System, Mol. Cell. Biol., 1987, vol. 7, pp. 379–387.
Dull, T., Zufferey, R., Kelly, M., et al., A Third-Generation Lentivirus Vector with a Conditional Packaging System, J. Virol., 1998, vol. 72, pp. 8463–8471.
Fossel, M.B., Cells, Aging, and Human Disease, New York: Oxford Univ., 2004.
Kang, H.J., Choi, Y.S., Hong, S.-B., et al., Ectopic Expression of the Catalytic Subunit of Telomerase Protects against Brain Injury Resulting from Ischemia and NMDA-Induced Neurotoxicity, J. Neurosci., 2004, vol. 24, pp. 1280–1287.
Kanzaki, Y., Onoue, F., Ishikawa, F., et al., Telomerase Rescues the Expression Level of Keratinocyte Growth Factor and Insulin-Like Growth Factor-II in Senescent Human Fibroblasts, Exp. Cell Res., 2002, vol. 279, pp. 321–329.
Kanzaki, Y., Onoue, F., Sakurai, H., et al., Telomerase Upregulates Expression Levels of Interleukin (IL)-1alpha, IL-1beta, IL-6, IL-8, and Granulocyte-Macrophage Colony-Stimulating Factor in Normal Human Fibroblasts, Biochem. Biophys. Res. Commun., 2003, vol. 305, pp. 150–154.
Lindvall, C., Hou, M., Komurasaki, T., et al., Molecular Characterization of Human Telomerase Reverse Transcriptase-Immortalized Human Fibroblasts by Gene Expression Profiling: Activation of the Epiregulin Gene, Cancer Res., 2003, vol. 63, pp. 1743–1747.
Mathew, S., Arandjelovic, S., Beyer, W.F., et al., Characterization of the Interaction between Alpha2-Macroglobulin and Fibroblast Growth Factor-2: The Role of Hydrophobic Interactions, Biochem. J., 2003, vol. 374, pp. 123–129.
Meyerson, M., Counter, C.M., Eaton, E.N., et al., HEST2, the Putative Human Telomerase Catalytic Subunit Gene, Is Up-Regulated in Tumor Cells and during Immortalisation, Cell, 1997, vol. 90, pp. 785–795.
Naldini, L., Blomer, U., Gallay, P., et al., In vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector, Science, 1996, vol. 272, pp. 263–267.
Narushima, M., Kobayashi, N., Okitsu, T., et al., A Human Beta-Cell Line for Transplantation Therapy to Control Type 1 Diabetes, Nat. Biotech., 2005, vol. 23, pp. 1274–1282.
Pear, W.S., Nolan, G.P., Scott, M.L., et al., Production of High-Titer Helper-Free Retroviruses by Transient Transfection, Proc. Natl. Acad. Sci. USA, 1993, vol. 90, pp. 8392–8396.
Rea, S.L., Wu, D., Cypser, J.R., et al., A Stress-Sensitive Reporter Predicts Longevity in Isogenic Populations of Caenorhabditis elegans, Nat. Genet., 2005, vol. 37, pp. 894–898.
Rieske, P., Krynska, B., and Azizi, S.A., Human Fibroblast-Derived Cell Lines Have Characteristics of Embryonic Stem Cells and Cells of Neuro-Ectodermal Origin, Differentiation, 2005, vol. 73, pp. 474–483.
Sablina, A.A., Budanov, A.V., Ilyinskaya, G.V., et al., The Antioxidant Function of the P53 Tumor Suppressor, Nat. Med., 2005, vol. 11, pp. 1306–1313.
Scadden, D.T., The Stem-Cell Niche as an Entity of Action, Nature, 2006, vol. 441, pp. 1075–1079.
Shevchenko, A., Wilm, M., Vorm, O., et al., Mass Spectrometric Sequencing of Proteins Silver-Stained Polyacrylamide Gels, Anal. Chem., 1996, vol. 68, pp. 850–858.
Torres, C., Lewis, L., and Cristofalo, V.J., Proteasome Inhibitors Shorten Replicative Life Span and Induce a Senescent-Like Phenotype of Human Fibroblasts, J. Cell Physiol., 2006, vol. 207, pp. 845–853.
Walter, M., Davies, J.P., and Ioannou, Y.A., Telomerase Immortalization Upregulates Rab9 Expression and Restores LDL Cholesterol Egress from Niemann-Pick C1 Late Endosomes, J. Lipid Res., 2003, vol. 44, pp. 243–253.
Yegorov, Ye.E., Terekhov, S.M., Vishnyakova, K.S., et al., Immortalization of Normal Fibroblasts of Adult Human Skin by Incorporation of the Gene of Telomerase Catalytic Component, Biol. Membr., 2002, vol. 19, pp. 483–490.
Yegorov, Ye.E., Terekhov, S.M., Vishnyakova, K.S., et al., Telomerization is a Way of Production of Immortal Human Cells Preserving Their Normal Properties, Ontogenez, 2003, vol. 34, pp. 183–192.
Yegorov, Ye.E., Moldaver, M.V., Terekhov, S.M., et al., Telomerization Does Not Enhance the Capacity of Human Fibroblasts to Colony Formation, Biol. Membr., 2004, vol. 21, pp. 298–305.
Yegorov, Ye.E., Moldaver, M.V., Vishnyakova, K.S., et al., Effects of Oxygen on Culture of Human Fibroblasts, Biol. Membr., 2005a, vol. 22, pp. 43–51.
Yeorov, Ye.E., Moldaver, M.V., Vishnyakova, K.S., et al., Evidence in Favor of Differentiation of Immortal Human Fibroblasts in vitro under Sparse Planting, Biol. Membr., 2005b, vol. 22, pp. 458–465.
Young, J.I., Sedivy, J.M., and Smith, J.R., Telomerase Expression in Normal Human Fibroblasts Stabilizes DNA 5-Methylcytosine Transferase I, J. Biol. Chem., 2003, vol. 278, pp. 19 904–19 908.
Zhang, H., Pan, R.H., and Cohen, S.N., Senescence-Specific Gene Expression Fingerprints Reveal Cell-Type-Dependent Physical Clustering of Up-Regulated Chromosomal Loci, Proc. Natl. Acad. Sci. USA, 2003, vol. 100, pp. 3251–3256.
Zhang, H., Herbert, B.S., Pan, R.H., et al., Disparate Effects of Telomere Attrition on Gene Expression during Replicative Senescence of Human Mammary Epithelial Cells Cultured under Different Conditions, Oncogene, 2004, vol. 23, pp. 6193–6198.