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The influence of INK4 proteins on growth and self-renewal kinetics of hematopoietic progenitor cells

The influence of INK4 proteins on growth and self-renewal kinetics of hematopoietic progenitor cells
The influence of INK4 proteins on growth and self-renewal kinetics of hematopoietic progenitor cells
This study investigated the influence of expression of proteins of the INK4 family, particularly p16, on the growth and self-renewal kinetics of hematopoietic cells. First, retrovirus-mediated gene transfer (RMGT) was used to restore p16(INK4a) expression in the p16(INK4a)-deficient lymphoid and myeloid cell lines BV173 and K562, and it was confirmed that this inhibited their growth. Second, to sequester p16(INK4a) and related INK4 proteins, cyclin-dependent kinase 4 (CDK4) was retrovirally transduced into normal human CD34(+) bone marrow cells and then cultured in myeloid colony-forming cell (CFC) assays. The growth of CDK4-transduced colonies was more rapid; the cell-doubling time was reduced; and, upon replating, the colonies produced greater yields of secondary colonies than mock-untransduced controls. Third, colony formation was compared by marrow cells from p16(INK4a-/-) mice and wild-type mice. The results from p16(INK4a-/-) marrow were similar to those from CDK4-transduced human CFCs, in terms of growth rate and replating ability, and were partially reversed by RMGT of p16(INK4a). Lines of immature granulocytic cells were raised from 15 individual colonies grown from the marrow of p16(INK4a-/-) mice. These had a high colony-forming ability (15%) and replating efficiency (96.7%). The p16(INK4a-/-) cell lines readily became growth factor-independent upon cytokine deprivation. Taken together, these results demonstrate that loss of INK4 proteins, in particular p16(INK4a), increases the growth rate of myeloid colonies in vitro and, more importantly, confers an increased ability for clonal expansion on hematopoietic progenitor cells.
in-vitro, cell division, in vitro, kinetics, non-U.S.gov't, cyclin-dependent kinase inhibitor p16, london, human, cell line, physiology, bone marrow, adult, growth, bone marrow cells, proteins, protein, bone, gene expression regulation, mice, time, humans, secondary, cytology, animals, family, tumor suppressor, hematopoietic stem cells, genes, research support
0006-4971
2604-2610
Lewis, J.L.
bedc80ed-a391-46e0-8292-619f6cd4e9f4
Chinswangwatanakul, W.
6f13befc-557f-443d-bbc5-1e6673d4ceba
Zheng, B.
ac30ead0-ef10-4214-a439-9aaaae8223ff
Marley, S.B.
52ceac57-e56c-43e6-9a03-b1745dccdc03
Nguyen, D.X.
87211257-99c2-413b-9ba5-4f0a480b0121
Cross, N.C.
6b31788a-9b23-4938-ace2-5610032377a3
Banerji, L.
0080ab0e-e060-410e-ba78-0af0b2380178
Glassford, J.
18f79714-267b-4146-81ae-653094c139a3
Thomas, N.S.
df2d7c6d-2c96-4aaa-a7ef-7f7987759cf4
Goldman, J.M.
c0abeb85-851b-4680-b28e-e646d94a91f7
Lam, E.W.
99986b07-0047-40e2-9e9e-99dfcc05b6e6
Gordon, M.Y.
30181c9e-ba2f-43d5-a0e3-d0a92c33d7c0
Lewis, J.L.
bedc80ed-a391-46e0-8292-619f6cd4e9f4
Chinswangwatanakul, W.
6f13befc-557f-443d-bbc5-1e6673d4ceba
Zheng, B.
ac30ead0-ef10-4214-a439-9aaaae8223ff
Marley, S.B.
52ceac57-e56c-43e6-9a03-b1745dccdc03
Nguyen, D.X.
87211257-99c2-413b-9ba5-4f0a480b0121
Cross, N.C.
6b31788a-9b23-4938-ace2-5610032377a3
Banerji, L.
0080ab0e-e060-410e-ba78-0af0b2380178
Glassford, J.
18f79714-267b-4146-81ae-653094c139a3
Thomas, N.S.
df2d7c6d-2c96-4aaa-a7ef-7f7987759cf4
Goldman, J.M.
c0abeb85-851b-4680-b28e-e646d94a91f7
Lam, E.W.
99986b07-0047-40e2-9e9e-99dfcc05b6e6
Gordon, M.Y.
30181c9e-ba2f-43d5-a0e3-d0a92c33d7c0

Lewis, J.L., Chinswangwatanakul, W., Zheng, B., Marley, S.B., Nguyen, D.X., Cross, N.C., Banerji, L., Glassford, J., Thomas, N.S., Goldman, J.M., Lam, E.W. and Gordon, M.Y. (2001) The influence of INK4 proteins on growth and self-renewal kinetics of hematopoietic progenitor cells. Blood, 97 (9), 2604-2610.

Record type: Article

Abstract

This study investigated the influence of expression of proteins of the INK4 family, particularly p16, on the growth and self-renewal kinetics of hematopoietic cells. First, retrovirus-mediated gene transfer (RMGT) was used to restore p16(INK4a) expression in the p16(INK4a)-deficient lymphoid and myeloid cell lines BV173 and K562, and it was confirmed that this inhibited their growth. Second, to sequester p16(INK4a) and related INK4 proteins, cyclin-dependent kinase 4 (CDK4) was retrovirally transduced into normal human CD34(+) bone marrow cells and then cultured in myeloid colony-forming cell (CFC) assays. The growth of CDK4-transduced colonies was more rapid; the cell-doubling time was reduced; and, upon replating, the colonies produced greater yields of secondary colonies than mock-untransduced controls. Third, colony formation was compared by marrow cells from p16(INK4a-/-) mice and wild-type mice. The results from p16(INK4a-/-) marrow were similar to those from CDK4-transduced human CFCs, in terms of growth rate and replating ability, and were partially reversed by RMGT of p16(INK4a). Lines of immature granulocytic cells were raised from 15 individual colonies grown from the marrow of p16(INK4a-/-) mice. These had a high colony-forming ability (15%) and replating efficiency (96.7%). The p16(INK4a-/-) cell lines readily became growth factor-independent upon cytokine deprivation. Taken together, these results demonstrate that loss of INK4 proteins, in particular p16(INK4a), increases the growth rate of myeloid colonies in vitro and, more importantly, confers an increased ability for clonal expansion on hematopoietic progenitor cells.

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More information

Published date: 2001
Keywords: in-vitro, cell division, in vitro, kinetics, non-U.S.gov't, cyclin-dependent kinase inhibitor p16, london, human, cell line, physiology, bone marrow, adult, growth, bone marrow cells, proteins, protein, bone, gene expression regulation, mice, time, humans, secondary, cytology, animals, family, tumor suppressor, hematopoietic stem cells, genes, research support

Identifiers

Local EPrints ID: 59988
URI: http://eprints.soton.ac.uk/id/eprint/59988
ISSN: 0006-4971
PURE UUID: 3f8ad445-8d1c-456a-b193-5bba50fc30cc

Catalogue record

Date deposited: 02 Sep 2008
Last modified: 22 Jul 2022 21:12

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Contributors

Author: J.L. Lewis
Author: W. Chinswangwatanakul
Author: B. Zheng
Author: S.B. Marley
Author: D.X. Nguyen
Author: N.C. Cross
Author: L. Banerji
Author: J. Glassford
Author: N.S. Thomas
Author: J.M. Goldman
Author: E.W. Lam
Author: M.Y. Gordon

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