Interactions of dimethyl sulfoxide and granulocyte‐macrophage colony‐stimulating factors on the growth and maturation of HL‐60 cells

Abstract

We have studied the interactions of dimethyl sulfoxide (DMSO), Giant Cell Tumor (GCT) cell-conditioned medium (GCT CM), and highly purified granulocyte-macrophage colony-stimulating factors (GM-CSF) on the growth and maturation of a highly passaged population of HL-60 cells. DMSO produced dose-dependent inhibition of HL-60 growth in liquid and semisolid media. Growth was partially to completely restored by the addition of GCT CM to cultures. Experiments in which cell volume, cell cycle kinetics, tritiated thymidine (3HTdr) incorporation, cell number, and nitroblue tetrazolium (NBT) reduction were compared during culture indicated that DMSO inhibited the spontaneous increase in cell volume and flow of cells through the cell cycle which occurred in the first day of culture, the increase in 3HTdr incorporation which was detectable by day 2; and the increment in cell counts which occurred by day 3. These effects were opposed by GCT CM. In contrast, the DMSO-induced increase in NBT reduction which occurred by day 6 was not influenced by GCT CM. The major principle opposing DMSO was GM-CSF, since (1) highly purified GM-CSF from GCT cells and recombinant GM-CSF from COS cells transfected with the Mo cell GM-CSF gene overcame greater than 50% of DMSO inhibition; and (2) conditioned media from cells not producing CSF, G-CSF from GCT cells, and recombinant G-CSF from Escherichia coli transfected with the G-CSF gene from 5,637 cells were inactive. DMSO had little or no effect on the elaboration of autostimulatory activity by HL-60 cells. DMSO is a useful agent for inhibiting the spontaneous growth of HL-60 cells and restoring their dependence on GM-CSF, a property which may be mediated through the effects of DMSO on cell cycle kinetics and/or maturation.

Multiple mechanisms for the inhibition of rRNA synthesis during HL‐60 leukemia cell differentiation

“…The ter stationary phase (Sollner-Webb and Tower, 1986; cellular changes that eventually lead to growth arrest Grummt et al, 1977;Grummt et al, 1976; Cavanaugh begin early in the differentiation program (Fibach et al, et al, 1984). Changes in rRNA transcription appear to 1982; von Melchner and Hoffken, 1985) and are partially be more closely related to the cell growth cycle than to reversible by the growth factor GM-CSF (Schwartz and the synthesis of DNA or the immediate entry of cells Maher, 1988;Brennan et al, 1987). into S phase (Sollner-Webb and Tower, 1986).…”

Interactions of dimethyl sulfoxide and granulocyte‐macrophage colony‐stimulating factor on the cell cycle kinetics and phosphoproteins of G1‐enriched HL‐60 cells: Evidence of early effects on lamin B phosphorylation

“…Two HL-60 populations were used in these studies: the highly passaged line, HL-60 UR, has been in continuous culture in this laboratory for 10 years, whereas HL-60 passage 76 cells were thawed from frozen stocks and used from passages 76-100. Both were maintained in McCoy's 5A Medium (GIBCO) with 10% fetal bovine serum (Hyclone) ("standard medium") and passaged twice weekly as previously described (Brennan et al, 1987). Cells from the late exponentialearly plateau phase of growth (-1 x lo6 cells/ml for passage 76 and -2 x 106 cells/ml for UR) were washed in buffer and resuspended in standard medium (growth studies), 3% serum-medium (elutriation), or phosphatefree, serum-free medium ([32Pl-orthophosphate labeling).…”

“…HL-60 cell proliferation and maturation 1 X lo5 cells/ml were cultured at 37°C in 25 cm2 tissue culture flasks containing standard medium, 0-100 uiml recombinant human GM-CSF (RH-CSF, Genzyme, specific activity 5 x lo7 CFU-GM/mg protein), and 0-1.5% DMSO (Fisher Scientific) for 7 days. Cell viability was assessed by trypan blue exclusion; cell concentrations and volume distributions were quantified with an electronic particle counter-sizer (Coulter), and maturation was assessed by nitroblue tetrazolium (NBT) reduction, as previously described (Brennan et al, 1987).…”

“…We have previouslv reported that DMSO inhibits the autonomous growth" of -unsynchronized, highly passaged HL-60 myeloid leukemia cells and that this growth inhibition is reversed by addition of recombi-nant human GM-CSF to cultures (Brennan et al, 1987). Since DMSO-induced granulocytic differentiation (NBT reduction) is not antagonized by GM-CSF, their conjoint presence in cultures produces a facsimile of normal development where renewal and differentiation are balanced.…”

All-trans retinoic acid induces p62DOK1 and p56DOK2 expression which enhances induced differentiation and G0 arrest of HL-60 leukemia cells

“…HL-60 cells are cytokine-dependent myelomonocytic precursor cells [31,32] that retain their bipotent ability to differentiate along either the myeloid or the monocytic lineages when treated with all-trans retinoic acid (atRA) or 1,25-dihydroxyvitamin D3(VD3), respectively [33][34][35]. They express an ensemble of atRA-or VD3-induced or regulated cell surface receptors such as the insulin receptor [36,37] and c-FMS receptor tyrosine kinases [38,39], FcgRIIA [40], BLR1 [41], and CD38 [42,43].…”