Abstract
Differentiation leads to the cessation of cellular proliferation, but little is known about the molecular mechanisms of growth arrest. We compared the effect of two differentiation inducers, 12-o-tetradecanoyl 13-acetate (TPA) and dimethyl sulfoxide (DMSO) on both the cell-cycle and the modulation of G2-related genes in synchronized HL60 cells. TPA treatment of HL60 cells resulted in G1 arrest within 24 h. In contrast, the cell cycling of DMSO-treated cells was initially accelerated and they progressed to the second cycle before accumulating in the G1 phase. Expression of cyclin B, cdc25, wee1 and cdc2 was studied during cell cycle arrest by Northern blot hybridization. Expression of cyclin B, cdc25 and cdc2 fluctuated in association with cell cycle progression towards the G2/M phase, while wee1 expression remained constant in untreated cells. These four genes were highly expressed in TPA-treated cells for the first 12 h, but drastic down-regulation was seen at 18 h and expression became undetectable after 24 h. In contrast, no remarked changes of gene expression were seen in DMSO-treated cells. These findings suggest that cell cycle progression along with the initial process of differentiation in response to TPA differs from the response to DMSO and that the down-regulation of cdc2 expression by TPA-treated HL60 cells contributes to endorsement of G1 arrest.
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Grosso LE, Pitot HC: Modulation of c-myc expression in HL60 cell line. Biochem Biophys Res Commun 119: 473–480, 1984
Gaynor R, Simon K, Koeffler P: Expression of c-jun during macrophage differentiation of HL-60 cells. Blood 77: 2618–2633, 1991
Cobrinik D, Dowdy SF, Hinds PW, Mittnacht S, Weinberg RA: The retinoblastoma protein and the regulation of cell cycling. TIBS 17: 312–315, 1992
Hihara 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
Akiyama T, Toyoshima K: Marked alteration in phosphorylation of the RB protein during differentiation of human promyelocytic HL60 cells. Oncogene 5: 179–183, 1990
Nurse P: Universal control mechanism regulating onset of M-phase. Nature 344: 503–508, 1990
Riabowol K, Draetta G, Brizuela L, Vandre D, Beach D: The cdc2 kinase is a nuclear protein that is essential for mitosis in mammalian cells. Cell 57: 393–401, 1989
Pines J, Hunter T: Isolation of a human cyclin cDNA: Evidence for cyclin mRNA and protein regulation in the cell cycle and for interaction with p34cdc2. Cell 58: 833–846, 1989
Gautier J, Minshull J, Lohka M, Glotzer M, Hunt T, Maller JL: Cyclin is a component of maturation-promoting factor from Xenopus. Cell 60: 487–494, 1990
Gautier J, Matsukawa T, Nurse P, Maller J: Dephosphorylation and activation of Xenopus p34cdc2 protein kinase during the cell cycle. Nature 339: 626–629, 1989
Morla AO, Draetta G, Beach D, Wang JYJ: Reversible tyrosine phosphorylation accompanies activation during entry into mitosis. Cell 58: 193–203, 1989
Krek W, Nigg EA: Differential phosphorylation of vertebrate p34cdc2 kinase at the G1/S and G2/M transitions of the cell cycle: Identification of major phosphorylation sites. EMBO J 10: 305–316, 1991
Lundgren K, Walworth N, Booher R, Dembski M, Kirschner M, Beach D: mik1 and wee1 cooperate in the inhibitory tyrosine phosphorylation of cdc2. Cell 64: 1111–1122, 1991
Kumagai A, Dunphy WG: The cdc25 protein controls tyrosine dephosphorylation of the cdc2 protein in a cell-free system. Cell 64: 903–914, 1991
Dunphy WG, Kumagai A: The cdc25 protein contains an intrinsic phosphatase activity. Cell 67: 189–196, 1991
Gautier J, Solomon MJ, Booher RN, Bazan JF, Kirschner MW: cdc25 is a specific tyrosine phosphatase that directly activates p34cdc2. Cell 67: 197–211, 1991
Galaktionov K, Beach D: Specific activation of cdc25 tyrosine phosphatases by B-type cyclins: Evidence for multiple roles of mitotic cyclins. Cell 67: 1181–1194, 1991
Jessus C, Beach D: Occilation of MPF is accompanied by periodic association between cdc25 and cdc2-cyclin B. Cell 68: 323–332, 1992
Lübbert M, Harrmann F, Koeffler HP: Expression and regulation of myeloid-specific genes in normal and leukemic myeloid cells. Blood 77: 909–924, 1991
Heintz N, Sive HL, Roeder RG: Regulation of human histone gene expression: Kinetics of accumulation and changes in the rate of synthesis and in the half-lives of individual histone mRNAs during the HeLa cell cycle. Mol Cell Biol 3: 539–550, 1983
Maniatis T, Fritsch EF, Sambrook J: Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, New York, 1982, pp 187–210
Yamada H, Sakamoto H, Taira M, Nishimura S, Shimosato Y, Terada M, Sugimura T: Amplifications 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 77: 370–375, 1986
Lee MG, Nurse P: Complementation used to clone a human homologue of the fission yeast cell cycle control gene cdc2. Nature 327: 31–35, 1987
Nagata A, Igarashi M, Jinno S, Suto K, Okayama H: An additional homolog of the fission yeast cdc25+ gene occurs in humans and is highly expressed in some cancer cells. The new biologist 3: 959–968, 1991
Igarashi M, Nagata A, Jinno S, Suto K, Okayama H: Wee1+-like gene in human cells. Nature 353: 80–83, 1991
Pardee AB: G1 events and regulation of cell proliferation. Science 246: 603–608, 1989
Murray AW, Kirschner MW: Dominoes and clocks: The union of two views of the cell cycle. Science 246: 614–621, 1989
McGowan CH, Russell P, Reed SI: Periodic biosynthesis of the human M-phase promoting factor catalytic subunit p34 during the cell cycle. Mol Cell Biol 10: 3847–3851, 1990
Draetta G, Luca F, Westendorf J, Brizuela L, Ruderman J, Beach D: cdc2 protein kinase is complexed with both cyclin A and B: Evidence for proteolytic inactivation of MPF. Cell 56: 829–838, 1989
Enoch T, Nurse P: Mutation of fission yeast cell cycle control genes abolishes dependence of mitosis on DNA replication. Cell 60: 665–673, 1990
Rowley R, Hudson J, Young PG: The wee1 protein kinase is required for radiation-induced mitotic delay. Nature 356: 353–355, 1992
Smythe C, Newport JW: Coupling of mitosis to the completion of S phase in Xenopus occurs via modulation of the tyrosine kinase that phosphorylates p34cdc2. Cell 68: 787–797, 1992
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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). https://doi.org/10.1007/BF00926850
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DOI: https://doi.org/10.1007/BF00926850