Abstract
Using a serum-free culture method, we investigated the effects of vitamin A on the proliferation of human distal airway epithelial cells. Outgrowth of epithelial cells from lung tissue explants was enhanced by treatment with all-trans retinol at concentrations of 10−8 to 10−7 M. The colony-forming activity of cells harvested from the primary culture and replated onto Swiss 3T3 fibroblastic feeders was, in contrast, significantly reduced by 10−7 M to 10−5 M retinol. When the primary cells were harvested and subcultured on Primaria plates, population expansion was also inhibited by retinol at 10−10 to 10−6 M. We further investigated the cells to determine whether there was any difference in sensitivity to the growth-inhibitory effects of vitamin A between cells from the primary culture incubated with and without retinol. The population increase in cells harvested from the primary culture was inhibited equally in retinol-treated and non-treated cells by subsequent treatment with retinol or retinoic acid, this inhibition being dose-dependent. DNA synthetic activity was also inhibited. Interestingly, both the growth rate and the colony-forming efficiency on feeders were greater in the subculture of cells from the retinol-treated primary culture than in those non-treated. When the cells in the secondary subculture were treated with retinoic acid and replated again, they showed a greater population increase rate than those non-treated. Our results showed that human distal airway epithelial cells isolated from lung tissue were sensitive to the growth-inhibitory effect of vitamin A, but the proliferative potential in some fraction of the epithelial cell population was possibly enhanced by vitamin A treatment.
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References
Chopra DP (1983) Cell dynamics in explants derived from tracheas of hamsters fed normally and vitamin A-deficient diets. Cell Tissue Kinet 16: 155–165
Chopra DP (1983) Retinoid reversal of squamous metaplasia in organ cultures of tracheas derived from hamsters-fed on vitamin A-deficient diet. Eur J Cancer Clin Oncol 19: 847–857
De Ruyter MC, De Leenheer AP (1976) Determination of serum retinol (vitamin A) by high-speed liquid chromatography. Clin Chem 22: 1593–1595
Doyle LA, Giangiulo D, Hussain A, Park H-J, Chiu Yen R-W, Borges M (1989) Differentiation of human variant small cell lung cancer cell lines to a classic morphology by retinoic acid. Cancer Res 49: 6745–6751
Emura M, Mohr U, Riebe M, Aufderheide M, Dungworth DL (1988) Regulation of growth and differentiation by vitamin A in a cloned fetal lung epithelial cell line cultured on collagen gel in hormone-supplemented medium. In Vitro Cell Dev Biol 24: 639–648
Jetten AM, Smits H (1985) Regulation of differentiation of tracheal epithelial cells by retinoids. In: Sporn MB (chairman) Retinoids, differentiation and disease, Ciba Foundation Symposium 113, Pitman, London, pp 61–76
Jetten AM, Brody AR, Deas MA, Hook GER, Rearick J, Thacher SM (1987) Retinoic acid and substratum regulate the differentiation of rabbit tracheal epithelial cells into squamous and secretory phenotype. Lab Invest 6: 654–664
Kitamura H, Shibagaki T, Inayama Y, Ito T, Kanisawa M (1990) Growth and differentiation of human distal airway epithelial cells in culture. Effects of small amounts of serum in defined medium. Lab Invest 63: 420–428
Klann RC, Marchok AC (1982) Effects of retinoic acid on cell proliferation and cell differentiation in a rat tracheal epithelial cell line. Cell Tissue Kinet 15: 473–482
Lechner JF, Haugen A, Autrup H, McClendon IA, Trump BF, Harris CC (1984) Clonal growth of epithelial cells from normal adult human bronchus. Cancer Res 41: 2294–2304
Lee T-C, Wu R, Brody AR, Barrett JC, Nettesheim P (1984) Growth and differentiation of hamster tracheal epithelial cells in culture. Exp Lung Res 6: 27–45
Lotan R (1980) Effects of vitamin A and its analogs (retinoids) on normal and neoplastic cells. Biochim Biophys Acta 605: 33–91
Marchok AC, Cone V, Nettesheim P (1975) Induction of squamous metaplasia (vitamin A deficiency) and hypersecretory activity in tracheal organ cultures. Lab Invest 33: 451–460
McDowell EM, Keenan KP, Huang M (1984) Effects of vitamin A-deprivation on hamster tracheal epithelium. A quantitative morphological study. Virchows Arch [B] 45: 197–219
McDowell EM, Kennan KP, Huang M (1984) Restoration of mucociliary tracheal epithelium following deprivation of vitamin A. A quantitative morphologic study. Virchows Arch [B] 45: 221–240
McDowell EM, Ben T, Coleman B, Chang S, Newkirk C, De Luca LM (1987) Effects of retinoic acid on the growth and morphology of hamster tracheal epithelial cells in primary culture. Virchows Arch [B] 54: 38–51
Mehta PP, Bertram JS, Loewenstein WR (1989) The actions of retinoids on cellular growth correlate with their actions on gap junctional communication. J Cell Biol 108: 1053–1065
Nervi C, Vollberg TM, George MD, Zelent A, Chambon P, Jetten AM (1991) Expression of nuclear retinoic acid receptors in normal tracheobronchial cells and lung carcinoma cells. Exp Cell Res 195: 163–170
Niles R, Kim C, Hyman B, Christensen T, Wasano K, Brody J (1988) Characterization of extended primary and secondary cultures of hamster tracheal epithelial cells. In Vitro 24: 457–463
Niles RM, Loewy BP, Brown K (1990) The effect of retinoic acid on growth and proto-oncogene expression in hamster tracheal epithelial cells. Am J Respir Cell Mol Biol 2: 365–371
Ochiai A, Emura M, Riebe-Imre M, Mohr U, Hilfrich J, Tahara E, Dungworth DL (1991) Secretory differentiation and cell type identification of a human fetal bronchial epithelial cell line (HFBE). Virchows Arch [B] 61: 217–226
Riaz-Ul-Haq, Pfahl M, Chytil F (1991) Retinoic acid affects the expression of nuclear retinoic acid receptors in tissues of retinol-deficient rats. Proc Natl Acad Sci USA 88: 8272–8276
Rutten AAJJL, Wilmer JWGM, Beems RB (1988) Effects of all-trans retinol and cigarette smoke condensate on hamster tracheal epithelium in organ culture. I. A cell proliferation study. Virchows Arch [B] 55: 167–175
Rutten AAJJL, Jongen WMF, de Haan LHJ, Hendriksen EGJ and Koeman JH (1988) Effect of retinol and cigarette-smoke condensate on dye-coupled intercellular communication between hamster tracheal epithelial cells. Carcinogenesis 9: 315–320
Shields PA, Jeffery PK (1987) The combined effects of vitamin A-deficiency and cigarette smoke on rat tracheal epithelium. Br J Exp Pathol 68: 705–717
Shimosato Y, Kodama T, Kameya T (1982) Morphogenesis of peripheral type adenocarcinoma of the lung. In: Shimosato Y, Melamed MR, Nettesheim P (eds) Morphogenesis of lung cancer, vol I, CRC press, Boca Raton, Florida, pp 65–90
Takizawa H, Beckmann JD, Shoji S, Claassen LR, Ertl RF, Linder J, Rennard SI (1990) Pulmonary macrophages can stimulate cell growth of bovine bronchial epithelial cells. Am J Respir Cell Mol Biol 2: 245–255
Takizawa H, Beckmann JD, Yoshida M, Romberger D, Rennard SI (1991) Regulation of bovine bronchial cell proliferation and protooncogene expression by growth factors. Am J Respir Cell Mol Biol 5: 548–555
Wu R, Nolan E, and Turner C (1985) Expression of tracheal differentiated functions in serum-free hormone-supplemented medium. J Cell Physiol 125: 167–18
Wu R, Yankaskas JR, Cheng E, Knowles MR, Boucher RC (1985) Growth and differentiation of human nasal epithelial cells in culture. Serum-free, hormone-supplemented medium and proteoglycan synthesis. Am Rev Respir Dis 132: 311–320
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Shibagaki, T., Ogata, T., Kitamura, H. et al. Effects of vitamin A on proliferation of human distal airway epithelial cells in culture. Vichows Archiv A Pathol Anat 424, 525–531 (1994). https://doi.org/10.1007/BF00191439
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DOI: https://doi.org/10.1007/BF00191439