Summary
Cyclopentenone prostaglandins (PGs) such as Δ12-PGJ2 and PGA are potent inducers of growth inhibition in a variety of cultured cells, including epidermal cells. These PGs are actively transported into cells by a specific carrier on cell membrane and accumulate in cell nuclei with binding to nuclear protein. To clarify the mechanism of cytotoxicity of these PGs in epidermal cells, we examined the effects of Δ12-PGJ2 on protein synthesis and cytoskeleton in the PAM 212 transformed mouse epidermal cell line. Cycloheximide at 1 Μg/ml culture medium exhibited a protective effect on cell growth inhibition of PAM 212 cells by Δ12-PGJ2. The analysis of cell lysate protein patterns by SDS-polyacrylamide gel electrophoresis revealed that 12-h incubation with Δ12-PGJ2 increased the amount of 70 kD protein in PAM 212 cells. The amount of 70 kD protein in Δ12-PGJ2-treated cells was markedly decreased by cotreatment with cycloheximide. This 70 kD protein was also induced in PAM 212 cells with treatment at 43‡C for 90 min, indicating that this synthesized protein belongs to the heat shock protein. The addition of Δ12-PGJ2 to confluent PAM 212 cells resulted in the disappearance of action filament, as visualized by fluorescent labeled phallacidine, but in contrast, keratin filament appeared to be intact during 12-h incubation with Δ12-PGJ2 at a concentration of 5 Μg/ml culture medium. These results suggest that the cytotoxicity of cyclopentenone PGs is at least in part due to induction of the synthesis of some protein(s), probably one of the heat shock proteins, and the damage to the actin filament in transformed cultured epidermal cells.
Similar content being viewed by others
References
Barak LS, Yocum RR, Nothnagel EA, Webb WW (1980) Fluorescence staining of the actin cytoskeleton in living cells with 7-nitrobenz-2-oxa-1,3-diazole-phallacidin. Proc Natl Acad Sci USA 77:980–984
Bhuyan BK, Adams EG, Badiner GJ, Li LH, Barden K (1986) Cell cycle effects of prostaglandins A1, A2, and D2 in human and murine melanoma cells in culture. Cancer Res 46:1688–1693
Bregman MD, Funk C, Fukushima M (1986) Inhibition of human melanoma growth by prostaglandin A, D, and J analogues. Cancer Res 46:2740–2744
Hirata Y, Hayashi H, Ito S, Kikawa Y, Ishibashi M, Sudo M, Miyazaki H, Fukushima M, Narumiya S, Hayaishi O (1986) Occurrence of 9-deoxy-Δ9,Δ12-13,14-dihydroprostaglandin D2 in human urine. J Biol Chem 263:16619–16625
Hughes-Fulford M, Wu J, Kato T, Fukushima M (1985) Inhibition of DNA synthesis and cell cycle by prostaglandins independent of cyclic AMP. In: Hayaishi O, Yamamoto S (eds) Advances in prostaglandin, thromboxane, and leukotriene research, vol 15. Raven Press, New York, pp 401–404
Ikai K, Imamura S (1988) Prostaglandin D2 in the skin. Int J Dermatol 27:141–149
Ikai K, Ujihara M, Kashihara M, Fukushima M (1987) Inhibition of the proliferation of transformed epidermal cells in culture by various prostaglandins. J Invest Dermatol 89:69–72
Kato T, Fukushima M, Kurozumi S, Noyori R (1986) Antitumor activity of Δ7-prostaglandin A1 and Δ12-PGJ2 in vitro and in vivo. Cancer Res 46:3538–3542
Kikuchi Y, Miyauchi M, Oomori K, Kita T, Kizawa I, Kato K (1986) Inhibition of human ovarian cancer cell growth in vitro and in nude mice by prostaglandin D2. Cancer Res 46:3364–3366
Kitajima Y, Inoue S (1985) Alteration in the arrangement of the keratin-type intermediate filaments during mitosis in cultured human keratinocytes. Eur J Cell Biol 38:219–225
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagents. J Biol Chem 193:265–275
Narumiya S, Fukushima M (1985) Δ12-Prostaglandin J2, an ultimate metabolite of prostaglandin D2 exerting cell growth inhibition. Biochem Biophys Res Commun 127:739–745
Narumiya S, Fukushima M (1986) Site and mechanism of growth inhibition by prostaglandins. I. Active transport and intracellular accumulation of cyclopentenone prostaglandins, a reaction leading to growth inhibition. J Pharmacol Exp Ther 239:500–505
Narumiya S, Ohno K, Fujiwara M, Fukushima M (1986) Site and mechanism of growth inhibition by prostaglandins. II. Temperature-dependent transfer of a cyclopentenone prostaglandin to nuclei. J Pharmacol Exp Ther 239:506–511
Narumiya S, Ohno K, Fukushima M, Fujiwara M (1987) Site and mechanism of growth inhibition by prostaglandins. III. Distribution and binding of prostaglandin A2 and Δ12-prostaglandin J2 in nuclei. J Pharmacol Exp Ther 242:306–311
Needleman P, Turk J, Jakschick BA, Morrison AR, Lefkowith JB (1986) Arachidonic acid metabolism. Annu Rev Biochem 55:69–102
Ohno K, Sakai T, Fukushima M, Narumiya S, Fujiwara M (1988) Site and mechanism of growth inhibition by prostaglandins. IV. Effect of cyclopentenone prostaglandins on cell cycle progression of G1-enriched HeLa S3 cells. J Pharmacol Exp Ther 245:294–298
Ohno K, Fukushima M, Fujiwara M, Narumiya S (1988) Induction of 68,000-dalton heat shock proteins by cyclopentenone prostaglandins. J Biol Chem 263:19764–19770
Ruzicka T (1988) The physiology and pathophysiology of eicosanoids in the skin. Eicosanoids 1:59–72
Sakai T, Yamaguchi B, Shiroki Y, Sekiguchi M, Fujii G, Nishino H (1984) Prostaglandin D2 inhibits the proliferation of human malignant tumor cells. Prostaglandins 27:17–26
Sakai T, Aoike A, Marui N, Kawai K, Nishino H, Fukushima M (1989) Protection by cycloheximide against cytotoxicity induced by vincristine, colchicine, or Δ12-prostaglandin J2 on human osteosarcoma cells. Cancer Res 49:1193–1196
Shimizu Y, Todo S, Imashuku S (1986) Cycloheximide reduced PGD2 or Δ12-PGJ2 cytotoxicity on NCG cells. Prostaglandins 32:517–525
Shimizu Y, Todo S, Imashuku S (1987) Selective synthesis and retention of 66k protein in a human neuroblastoma cell line (NCG) treated with a cytotoxic dosage of Δ12-prostaglandin J2. Prostaglandins 34:769–781
Terranova VP, Rohrbach DH, Martin CR (1980) Role of laminin in the attachment of PAM 212 (epithelial) cells to basement membrane collagen. Cell 22:719–726
Todo S, Hashida T, Shimizu Y, Imashuku S, Takamatsu T, Fujita S (1986) Cell kinetic studies of PGD2 cytotoxicity on the in vitro growth of human neuroblastoma. Prostaglandins Leukotrienes Med 23:55–65
Yuspa SH, Hawley-Nelson P, Koehler B, Stanley JR (1980) A survey of transformation markers in differentiating epidermal cell lines in culture. Cancer Res 40:4694–4703
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ikai, K., Fukushima, M. Effects of cytotoxic prostaglandin, Δ12PGJ2 on protein synthesis and cytoskeleton in transformed epidermal cells in culture. Arch Dermatol Res 282, 131–134 (1990). https://doi.org/10.1007/BF00493472
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00493472