Summary
Double histochemical staining followed by flow cytometric analysis was performed to determine whether involucrin synthesis is associated with a particular phase of the cell cycle. In low-calcium medium (0.1 mM) monolayer cultures the expression of involucrin was confined to about 8% of the total cell populations. When the concentration of calcium was increased to 1.8 mM, the percentage increased and the distribution pattern of the cell cycle changed. The addition of retinoic acid at concentrations in the range 10−8 M to 10−6 M to a high-calcium medium induced a further increase. The greatest increase in involucrin expression (up to five-fold) occurred in both the G1/G0 and G2 + M phases, while S phase cells showed a two-fold increase. The results indicate that involucrin synthesis is induced by retinoic acid, which occurs at any stage of the cell cycle, even in the S phase.
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References
Cline PR, Rice RH (1983) Modulation of involucrin and envelope competence in human keratinocytes by hydrocortisone, retinyl acetate and growth arrest. Cancer Res 43: 3203 to 3207
Dion LD, Gifford GE (1980) Retinoic acid induces a G1 cell cycle block in HeLa cell. Proc Soc Exp Biol Med 163: 510–514
Dover R, Watt FM (1987) Measurement of the rate of epidermal terminal differentiation: expression of involucrin by S-phase keratinocytes in culture and in psoriatic plaques. J Invest Dermatol 89: 349–352
Floyd EE, Jetten AM (1989) Regulation of type 1 (epidermal) transglutaminase mRNA levels during differentiation: down regulation by retinoid. Mol Cell Biol 9: 4846–4851
Green H, Watt FM (1982) Regulation by vitamin A of envelope crosslinking in cultured keratinocytes derived from different human epithelia. Mol Cell Biol 2: 1115–1117
Haddox MK, Russell DH (1979) Cell cycle specific locus of vitamin A inhibition of growth. Cancer Res 39: 2476–2480
Hennings H, Michael D, Cheng C et al. (1980) Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell 19: 245–254
Hohl D, Mehrel T, Lichti U et al. (1991) Characterization of human loricrin. J Biol Chem 266: 6626–6636
Mehrel T, Hohl D, Rothnagel JA et al. (1990) Identification of a major keratinocyte cell envelope protein, loricrin. Cell 61: 1103–1112
Pittelkow MR, Scott RE (1986) New techniques for the in vitro culture of human skin keratinocytes and perspectives on their use for grafting of patients with extensive burns. Mayo Clin Proc 61: 771–777
Regnier M, Vaigot P, Darmon M et al. (1986) Onset of epidermal differentiation in rapidly proliferating basal keratinocytes. J Invest Dermatol 87: 472–476
Rice RH, Green H (1979) Presence in human epidermal cells of a soluble protein precursor of the cross-linked envelope: activation of cross linking by calcium ions. Cell 18: 681–694
Varani J, Nickoloff BJ, Dixit VM et al. (1989) All-trans retinoic acid stimulates growth of adult human keratinocytes cultured in growth-factor deficient medium, inhibits production of thrombospondin and fibronectin, and reduces adhesion. J Invest Dermatol 93: 449–454
Watt FM (1984) Selective migration of terminally differentiating cell from the basal layer of cultured human epidermis J. Cell Biol 98: 16–21
Watt FM (1987) Influence of cell shape and adhesiveness on stratification and terminal differentiation of human keratinocytes in culture. J Cell Sci Suppl 8: 313–316
Watt FM, Green H (1982) Stratification and terminal differentiation of cultured epidermal cells. Nature 295: 434–436
Yaar M, Stanley JR, Katz SI (1981) Retinoic acid delays the terminal differentiation of keratinocytes in suspension culture. J Invest Dermatol 76: 363–366
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Nakatani, S., Okada, N., Okumura, H. et al. A two-colour flowcytometric study of cell kinetics and differentiation of human keratinocytes in culture. Arch Dermatol Res 284, 349–352 (1992). https://doi.org/10.1007/BF00372038
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DOI: https://doi.org/10.1007/BF00372038