Abstract
Differentiation induction by 12-o-tetradecanoyl 13-acetate (TPA) results in the growth arrest of HL60 cells in the G1 phase. However, little is known about the changes of cell cycle-regulating genes during this differentiation process. We investigated the changes of mRNA for various cyclins (A, C, D1, D2, D3 and E) and cdk2. Synchronized HL60 cells began to proliferate immediately after release from cell cycle block and cell cycle synchrony was obvious until the second S phase. TPA-treated cells accumulated in G1 phase within 24 h and most of the cells were arrested in this phase at 36 h. The expression of cyclins and cdk2 was studied by Northern blot hybridization or the reverse-transcription polymerase chain reaction (RT-PCR). TPA treatment altered the expression of all genes studied. The expression of cdk2 and cyclin A mRNA was markedly down-regulated. Cyclin E mRNA expression was also prominently down-regulated from 12 h to 36 h, at which time a second increase of its expression was observed in control cells. In contrast, the expression of cyclin D1 mRNA was induced by TPA, while its expression in control cells was undetectable by Northern blot hybridization throughout the cell cycle. Cyclin C expression was faint and fluctuated irrelevant of cell cycle, but its expression in both control and TPA-treated cells was higher than at baseline. Cyclin D2 expression remained stable in control cells and TPA treatment resulted in slight down-regulation at 12 h, but no difference was observed after 24 h. Cyclin D3 mRNA expression was slightly induced at 6 h, a time when its expression was down-regulated in control cells. At 48 h, these cyclins (C, D2, and D3) showed almost same level of expression as the control. These findings suggest that the down-regulation of cyclin A and cdk2 expression contributes to the G1 arrest of HL60 cells during monocytic differentiation induced by TPA and that cyclin D1 plays an additional role other than the regulation of cell cycle progression.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Avvisati G, Cate JW, Mandelli F: Acute promyelocytic leukaemia. Br J Haemat 81: 315–320, 1992
Leglise MC, Dent GA, Ayscue LH, Ross DW: Leukemic cell maturation: Phenotypic variability and oncogene expression in HL60 cells: A review. Blood Cells 13: 319–337, 1988
Nishizuka Y: The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature 334: 661–665, 1988
Sherr CJ: Mammalian G1 cyclins. Cell 73: 1059–1065, 1993
Pardee AB: G1 events and regulation of cell proliferation. Science 246: 603–608, 1989
Horiguchi-Yamada J, Yamada H: Differing responses of G2-related genes during differentiation of HL60 cells induced by TPA or DMSO. Mol Cell Biochem 119: 29–34, 1993
Maniatis T, Fritsch EF, Sambrook J: Molecular cloning: A Laboratory manual. Cold Spring Harbor Laboratory, New York, 1982, pp 187–210
Southern EM: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517, 1975
Yamada H, Sakamoto H, Taira M, Nishimura S, Shimosato Y, Terada M, Sugimura S: Amplification of both c-Ki-ras with a point mutation and c-myc in a primary pancreatic cancer and its metastatic tumors in lymph nodes. Jpn J Cancer Res (Gann) 77: 370–375, 1986
Ninomiya-Tsuji J, Nomoto S, Yasuda H, Reed SI, Matsumoto K: Cloning of a human cDNA encoding a CDC2-related kinase by complementation of a budding yeast cdc28 mutation. Proc Natl Acad Sci USA 88: 9006–9010, 1991
Pines J, Hunter T: Human cyclin A is adenovirus E1A-associated protein p60 and behaves differently from cyclin B. Nature 346: 760–763, 1990
Lew DJ, Dulic V, Reed SI: Isolation of three novel human cyclins by rescue of G1 cyclin (Cln) function in yeast. Cell 66: 1197–1206, 1991
Motokura T, Bloom T, Kim HG, Jüppner H, Ruderman JV, Kronenberg HM, Arnold A: A novel cyclin encoded by a bcl1-linked candidate oncogene, Nature 350: 512–515, 1991
Inaba T, Matsushime H, Valentine M, Roussel MF, Sherr CJ, Look AT: Genomic organization, chromosomal location, and independent expression of human cyclin D genes. Genomics 13: 565–574, 1992
Surmacz E, Reiss K, Sell C, Baserga R: Cyclin D1 messenger RNA is induced by platelet-derived growth factor in cultured fibroblasts. Cancer Res 52: 4522–4525, 1992
Soubeyran P, Cabanillas F, Lee MS: Analysis of the expression of the hybrid gene bel-2/IgH in follicular lymphomas. Blood 81: 122–127, 1993
Chen P, Scully P, Shew J, Wang JYJ, Lee W: Phosphorylation of the retinoblastoma gene product is modulated during the cell cycle and cellular differentiation. Cell 58: 1193–1198, 1989
Mihara K, Cao X, Yen A, Chandler S, Driscoll B, Murphree AL, T'Ang A, Fung YT: Cell cycle-dependent regulation of phosphorylation of the human retinoblastoma gene product. Science 246: 1300–1303, 1989
Lin BT, Gruenwald S, Morla AO, Lee W, Wang JY: Retinoblastoma cancer suppressor gene product is a substrate of the cell cycle regulator cdc2 kinase. EMBO J 10: 857–864, 1991
Lees JA, Buchkovich KJ, Marshak DR, Anderson CW, Harlow E: The retinoblastoma protein is phosphorylated on multiple sites by human cdc2. EMBO J 10: 4279–4290, 1991
Koff A, Cross F, Fisher A, Schumacher J, Leguellec K, Philippe M, Roberts JM: Human cyclin E, a new cyclin that interacts with two members of CDC2 gene family. Cell 66: 1217–1228, 1991
Xiong Y, Connolly T, Futcher B, Beach D: Human D-type cyclin. Cell 65: 691–699, 1991
Ewen ME, Shuss HK, Sherr CJ, Matsushime H, Kato J, Livingston DM: Functional interactions of the retinoblastoma protein with mammalian D-type cyclins. Cell 73: 487–497, 1993
Hinds PW, Mittnacht S, Dulic V, Arnold A, Reed SI, Weinberg RA: Regulation of retinoblastoma protein functions by expression of human cyclins. Cell 70: 993–1006, 1992
Matsushime H, Roussel MF, Sherr CJ: Novel mammalian cyclins (CYL Genes) expressed during G1. Cold Spring Harbor Symp Quant Biol 56: 69–74, 1991
Geng Y, Weinberg RA: Transforming growth factor β effects on expression of G1 cyclins and cyclin-dependent protein kinases. Proc Natl Acad Sci USA 90: 10315–10319, 1993
Ando K, Ajchenbaum-Cymbalista F, Griffin JD: Regulation of G1/S transition by cyclins D2 and D3 in hematopoietic cells. Proc Natl Acad Sci USA 90: 9571–9575, 1993
Lotem J, Cragoe EJ, Sachs L: Rescue from programmed cell death in leukemic and normal myeloid cells. Blood 78: 953–960, 1991
Koff A, Giordano A, Desai D, Yamashita K, Harper JW, Elledge S, Nishimoto T, Morgan DO, Franza BR, Roberts JM: Formation and activation of a cyclin E-cdk2 complex during the G1 phase of the human cell cycle. Science 257: 1689–1694, 1992
Bandara LR, Asdamczewski JP, Hunt T, Thangue NBL: Cyclin A and the retinoblastoma gene product complex with a common transcription factor. Nature 352: 249–251, 1991
Shirodkar S, Ewen M, DeCaprio JA, Morgan J, Livingston DM, Chittenden T: The transcription factor E2F interacts with the retinoblastoma product and a p107-cyclin A complex in a cell cycle-regulated manner. Cell 68: 157–166, 1992
Pagano M, Draetta G, Jansen-Dürr P: Association of cdk2 kinase with the transcription factor E2F during S phase. Science 255: 1144–1147, 1992
Nevins JR: EZF: A link between the Rb tumor suppressor protein and viral oncoproteins. Science 258: 424–429, 1992
Pagano M, Pepperkok R, Verde F, Ansorge W, Draetta G: Cyclin A is required at two points in the human cell cycle. EMBO J 11: 961–971, 1992
Meyerson M, Enders GH, Wu C, Su L, Gorka C, Nelson C, Harlow E, Tsai L: A family of human cdc2-related protein kinases. EMBO J 11: 2909–2917, 1992
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Horiguchi-Yamada, J., Yamada, H., Nakada, S. et al. Changes of G1 cyclins, cdk2, and cyclin A during the differentiation of HL60 cells induced by TPA. Mol Cell Biochem 132, 31–37 (1994). https://doi.org/10.1007/BF00925672
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00925672