Erythroleukemia cell lines hel and K‐562: Changes in isoenzyme profiles and morphology during induction of differentiation

Abstract

The effects of the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) and of hemin on the human erythroleukemia cell lines HEL and K-562 were studied using the parameters cellular morphology, cell proliferation, positivity in the nitro-blue tetrazolium (NBT)-reduction test, adherence to surface, development of pseudopodia, and isoenzyme profiles of carboxylic esterase, acid phosphatase, hexosaminidase and lactate dehydrogenase (LDH) separated by isoelectric focusing on horizontal polyacrylamide gels. TPA and hemin induced similar morphological changes in HEL and K-562, erythrocyte-like cells were seen to be the predominant population after exposure to hemin. Both TPA and hemin led to cessation of cell proliferation and to a dose-dependent decrease in the number of viable nucleated cells. Whereas HEL cells treated with TPA became positive in the NBT-reduction test, adhered to plastic surface and showed pseudopodia, HEL cells exposed to hemin and K-562 cells cultured with TPA or hemin did not show these markers indicative of the macrophage-cell lineage. TPA-treated HEL cells exhibited isoenzyme patterns which were characteristically seen in cells of the monocyte-macrophage complex. 'Myeloid isoenzyme profiles' were found in TPA-treated K-562 cells. As evidenced by the above described phenotypical changes, HEL cells appeared to differentiate along the macrophage and erythroid cell lineages. Induced K-562 cells displayed myeloid and erythrocytic markers.

Establishment of an erythroid cell line from primary CD36+ erythroid progenitor cells

“…Hemin, DMSO, and PMA are chemicals known to induce differentiation of various hematopoietic cell lines. CD36E cells treated with hemin, PMA, or DMSO did not exhibit the desired result of differentiation as reported with some erythroleukemic cell lines, such as K562 [22,25], HEL [25] UT7 [15], U937 [24], and HL-60 [24]. DMSO rather negatively affected cell viability.…”

PMA‐induced megakaryocytic differentiation of HEL cells is accompanied by striking modifications of protein kinase C catalytic activity and isoform composition at the nuclear level

“…Several pluripotent haemopoietic cell lines are able to differentiate towards the megakaryocytic phenotype following treatment with PMA or other chemical compounds (Lozzio & Lozzio, 1975;Martin & Papayannopoulou, 1982;Tabilio et al, 1984;Drexler et al, 1986;Marks et al, 1987;Alitalo et al, 1990;Long et al, 1990;Dai & Murphy, 1993;Murate et al, 1993). This suggests that the genetic defects responsible for the leukaemic phenotype are at least partially reversible.…”

Hemoglobin as a Direct Inhibitor of Cartilage Growth In Vitro

“…The Hb-induced growth inhibition can be attributed to the contained heme. Heme proteins have been implicated as growth inhibitors of hamster ovary cells (Lin, Al-Dakan and Gibson 1986), erythroleukemic cells (Drexler, Gaedicke and Minowada 1986), cell membranes (Shaklai, Avissar, Rabizadeh and Shaklai 1986), and normal hematopoetic cells (Malik, Agam and Djaldetti 1979). The mechanism of heme action remains speculative at this time; although we can postulate it induces oxidant stress at the tissue level, this remains to be elucidated.…”