Modulation of phorbol ester-induced HL-60 differentiation by prostaglandin E2.
When treated with phorbol tumor promoters, HL-60 cells undergo terminal differentiation evidenced by a transition from a non-phagocytic suspension culture to an attached fibroblast-like culture with high phagocytic activity. Internalization of fluorescent particles by cells exhibiting the phagocytic positive phenotype (phag+) provides a sensitive indication of promoter-induced differentiation, and the resulting fluorescent cells can be quantitatively analyzed by flow cytometry. The current study was initiated to further test the predictive power of a flow cytometry based HL-60 differentiation assay in the detection of agents associated with tumor promotion. Specifically, experiments were designed to assess the sensitivity of the test system to co-promoters which enhance promoter activity in vivo. Prostaglandin E2 (PGE2) was chosen as a model co-promoter since it has been shown to potentiate phorbol ester (i.e. 12-O-tetradecanoyl phorbol-13-acetate; TPA) induced biological effects in vivo. Results detailed in the current report indicate that PGE2 enhances TPA-induced differentiation of HL-60 cells in a dose-dependent manner. As with in vivo co-promotion experiments, PGE2 exhibited a maximum potentiating effect when administered prior to TPA. These data indicate that HL-60 cells are not only sensitive to phorbol promoters, but also to the co-promoter PGE2. These experiments support the hypothesis that a flow cytometry based HL-60 assay may prove useful for studying chemical agents or intrinsic cellular factors that are involved in the tumor promotion phase of carcinogenesis.
Dertinger
S D
SD
Litron Laboratories, Rochester, NY 14620, USA.
Torous
D K
DK
Tometsko
A M
AM
D015197:Carcinogenicity Tests; D002273:Carcinogens / Q000633:toxicity*; D002454:Cell Differentiation / Q000187:drug effects*; D002469:Cell Separation; D002471:Cell Transformation, Neoplastic / Q000187:drug effects*; D015232:Dinoprostone / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005347:Fibroblasts; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007938:Leukemia; D010586:Phagocytes; D013755:Tetradecanoylphorbol Acetate / Q000633:toxicity*; D014407:Tumor Cells, Cultured / Q000187:drug effects
D002273:Carcinogens; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate