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Transformed human bronchial epithelial cells (beas-2b) alter the growth and morphology of normal human bronchial epithelial cells in vitro

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Normal human bronchial epithelial cells (BE) and adenovirus-12 SV40 hybrid virus transformed, non-tumorigenic human bronchial epithelial cells (BEAS-2B) were cultured for 7 days in a serum free hormone supplemented medium. BE cells after 3 days in culture were exposed to conditioned medium (CMt) from confluent BEAS-2B cells. By day 7, CMt-treated BE cells exhibited a lower colony forming efficiency (CFE), fewer cells per colony, and a reduced mitotic index (MI) and BrdU (bromodeoxyuridine) labeling index. CMt also enhanced the expression of a terminally differentiated squamous phenotype in BE cells. Cell free lysates from BEAS-2B cells (CFLt) had effects similar to CMt on the MI and morphology of BE cells. In contrast, CMt and CFLt did not inhibit the growth, or alter the morphology of BEAS-2B cells. Conditioned medium from BE cells (CMn) did not reduce the growth of BEAS-2B cells, and had little effect on the morphology of BE cells. In co-cultures

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Abbreviations

BE:

normal bronchial epithelial cells

BEAS-2B:

adenovirus-12 SV40 hybrid virus transformed bronchial epithelial cells

CMn :

conditioned medium from BE cells

CMt :

conditioned medium from BEAS-2B cells

CFn :

cell free lysate from BE cells

CFLt :

cell free lysate from BEAS-2B cells

BrdU:

bromodeoxyuridine

KGM:

keratinocyte growth medium

TGF-β:

transforming growth factor type β

NCI-LHC:

National Cancer Institute-Laboratory of Human Carcinogenesis

References

  • ALBRIGHT, C.D., HAY, R., JONES, R.T. and RESAU, J.H. (1989). Discrimination of normal and transformed cells in vitro by cytologic and morphologic analysis. Cytotechnology 2:187–201.

    Google Scholar 

  • ALBRIGHT, C.D., KEENAN, K.P., COLOMBO, K.L. and RESAU J.H. (1990). Morphologic identification of epithelial cell types of the human tracheo-bronchus in cell culture. J. Tissue Culture Methods, in press.

  • AUERBACH, O., GERE, J.B. and FORMAN, J.B. (1957). Changes in the bronchial epithelium in relation to smoking and cancer of the lung. N. Engl. J. Med. 256: 97–102.

    Google Scholar 

  • BARRETT, L.A., McDOWELL, E.M., FRANK, A.L., HARRIS, C.C. and TRUMP, B.F. (1976). Long-term culture of human bronchial epithelium. Cancer Res. 36: 1003–1010.

    Google Scholar 

  • BONFIL, R.D., REDDELL, R.R., URA, H., REICH, R., FRIDMAN, R., HARRIS, C.C. and KLEIN-SZANTO, A.J.P. (1989). Invasive and metastatic potential of a vHa-ras-transformed human bronchial epithelial cell line. J. Natl. Cancer Inst. 81: 587–594.

    Google Scholar 

  • BONNARD, C., LECHNER, J.F., GERWIN, B.I., FUJIKI, H. and HARRIS, C.C. (1988). Effects of palytoxin or oubain on growth and squamous differentiation of human bronchial epithelial cells in vitro. Carcinogenesis 9:2245–2249.

    Google Scholar 

  • BOONE, C.W., SANFORD, K.K., FROST, J.K., MANTEL, N., GILL, G.W. and JONES, G.M. (1986). Cytomorphologic evaluation of the neoplastic potential of 28 cell culture lines by a panel of diagnostic cytopathologists. Int. J. Cancer 38:361–367.

    Google Scholar 

  • BYERS, T., VENA, J.E., METTLIN, C., SWANSON, M. and GRAHAM, S. (1984). Dietary vitamin A and lung cancer risks: An analysis by histologic subtypes. Am. J. Epidemiology 120:769–776.

    Google Scholar 

  • CAVENY, S. (1985). The role of gap junctions in development. Ann. Rev. Physiol. 47: 319–335.

    Google Scholar 

  • CHANG, L.-Y., WU, R. and NETTESHEIM, P. (1985). Morphological changes in rat tracheal cells during the adaptive and early growth phase in primary cell culture. J. Cell Sci. 74:283–301.

    Google Scholar 

  • CHOPRA, D.P., SULLIVAN, J., WILLE, J. and SIDDIQUI, K.M. (1987). Propagation of differentiating normal human tracheobronchial epithelial cells in serum-free medium. J. Cell Physiol. 130:173–181.

    Google Scholar 

  • DANIELPOUR, D., DART, L.L., FLANDERS, K.C., ROBERTS, A.B. and SPORN, M.G. (1989). Immunodetection and quantitation of the two forms of transforming growth factor-beta (TGF-β1 and TGF-β2) secreted by cells in culture. J. Cell Physiol. 138: 79–86.

    Google Scholar 

  • DAVIDSON, J.S., BAUMGARTEN, I.M. and HARLEY, E.H. (1985). Studies on the mechanism of phorbol ester-induced inhibition of intercellular junctional communication. Carcinogenesis 6:1353–1358, 1985.

    Google Scholar 

  • DE BUYSSCHER, E., KENNEDY, J. and MENDICINO, J. (1984). Synthesis of mucin glycoproteins by epithelial cells isolated from swine trachea by specific proteolysis. In Vitro Cell Dev. Biol. 20:433–446.

    Google Scholar 

  • DE MELLO, W.C. (1976). Modulation of junctional permeability. Fed Proc 43: 2692–2696, 1984. In: Mortality in Relation to Smoking: 20 Years' Observation on Male British Doctors (R. Doll and R. Peto, eds.). Br. Med. J. 32:1525–1536.

    Google Scholar 

  • DVORAK, H.F. (1986). Tumors: Wounds that do not heal. N. Engl. J. Med. 315: 1650–1659.

    Google Scholar 

  • FILDERMAN, A.E., SHAW, C. and MATTHAY, R.A. (1986). Etiology, pathology, natural history, manifestations, and diagnostic techniques. Invest. Radiol. 21:80–90.

    Google Scholar 

  • FROST, J.K. (1986). The Cell in Health and Disease. S. Karger, Basel.

    Google Scholar 

  • FROST, J.K., EROZAN, Y.S., GUPTA, P.K. and CARTER, D. (1984). Cytopathology. In: Early Lung Cancer, pp. 33–76. Igaku-Shoin, Inc., New York.

    Google Scholar 

  • FURCHT, L.T. (1986). Critical factors controlling angiogenesis: Cell products, cell matrix and growth factors. Lab Invest. 55:505–509.

    Google Scholar 

  • GAFFNEY, M. and ALTSHULER, B. (1988). Examination of the role of cigarette smoke in lung carcinogenesis using multistage models. J. Natl. Cancer Inst. 80:925–931.

    Google Scholar 

  • GRATZNER, H.C. (1982). Monoclonal antibody to 5-bromo- and 5-iododeoxy-uridine: A new reagent for detection of DNA replication. Science 218:474–475.

    Google Scholar 

  • HARRIS, H. and BRAMWELL, M.E. (1987). The suppression of malignancy by terminal differentiation: Evidence from hybrids between tumor cells and keratinocytes. J. Cell Sci. 87:383–388.

    Google Scholar 

  • HOUCK, P.C. (1976). Putative bronchial chalone. In: Chalones (J.D. Houck, ed.), pp. 394–400.: North-Holland and American Elsevier, Amsterdam.

    Google Scholar 

  • INAYAMA, Y., HOOK, G.E.R., BRODY, A.R., CAMERON, G.S., JETTEN, A.M., GILMORE, L.B., GRAY, T. and NETTESHEIM, P. (1988). The differentiation potential of tracheal basal cells. Lab. Invest. 58:706–717.

    Google Scholar 

  • INAYAMA, Y., HOO, G.E., BRODY, A.R., JETTEN, A.M., GRAY, T., MAHLER, J. and NETTESHEIM, P. (1989). In vitro and in vivo growth and differentiation of clones of tracheal basal cells. Am. J. Pathol. 134:539–549.

    Google Scholar 

  • JANIK, P., ASSOF, A. and BERTRAM, J.S. (1980). Inhibition of growth of primary and metastatic Lewis lung carcinoma cells by the phophodiesterase inhibitor isobutylmethylxanthine. Cancer Res. 40:1950–1954.

    Google Scholar 

  • JEFFERY, P.K. (1983). Morphologic features of airway surface epithelial cells and glands. Am. Rev. Resp. Dis. 128:S14-S20.

    Google Scholar 

  • JOHNSTON, W.W. (1986). Cytologic diagnosis of lung cancer: Principles and problems. Path. Res. Pract. 181:1–36.

    Google Scholar 

  • JONES, R.T. and ELLIGET, K.E. (1987). Method for the culture of human bronchial epithelial cells from tissues obtained at post-mortem intervals. In Vitro Cell Dev. Biol. 23:36A.

  • KE, Y., REDDEL, R.R., GERWIN, B.I., MIYASHITA, M., McMENAMIN, M., LECHNER, J.F. and HARRIS, C.C. (1988). Human bronchial epithelial cells with integrated SV40 virus T-antigen genes retain the ability to undergo squamous differentiation. Differentiation 38:60–66.

    Google Scholar 

  • KEENAN, K.P., SAFFIOTTI, U., STINSON, S.F., RIGGS, C.W. and McDOWELL, E.M. (1989). Multifactorial hamster respiratory carcinogenesis. Interdependent effects of cannula-induced mucosal wounding, saline, ferric oxide, benzo[a]pyrene and N-methyl-N-nitrosourea. Cancer Res. 49:1528–1540.

    Google Scholar 

  • LECHNER, J.F., HAUGEN, A., AUTRUP, H., McCLENDON, I.A., TRUMP, B.F. and HARRIS, C.C. (1981). Clonal growth of epithelial cells from normal adult human bronchus. Cancer Res. 41:2294–2304.

    Google Scholar 

  • LECHNER, J.F., HAUGEN, A., McCLENDON, I.A. and PETTIS, E.W. (1982). Clonal growth of normal adult human bronchial epithelial cells in serum-free medium. In Vitro Cell Dev. Biol. 18:633–642.

    Google Scholar 

  • LECHNER, J.F. and KAIGN, M.E. (1979). Application of the principles of enzyme kinetics to clonal growth rate assays: An approach for delineating interactions among growth promoting agents. J. Cell Physiol. 100:519–530.

    Google Scholar 

  • LECHNER, J.F., McCLENDON, I.A., LA VECK, M.A., SHAMSUDDIN, A.M. and HARRIS, C.C. (1983). Differential control by platelet factors of squamous differentiation in normal and malignant human bronchial epithelial cells. Cancer Res. 43:5915–5921.

    Google Scholar 

  • LIKE, B. and MASSAGUé, J. (1986). The anti-proliferative effect of type beta transforming growth factor occurs at a level distal from receptors for growth-activating factors. J. Biol. Chem. 261:13426–13429.

    Google Scholar 

  • LOEB, L.A. (1989). Endogenous carcinogenesis: Molecular oncology into the twenty-first century—presidential address. Cancer Res. 49:5489–5496.

    Google Scholar 

  • LOEB, L.A., ERNSTER, V.L., WARNER, K., ABBOTTS, J. and LASZLO, J. (1984). Smoking and lung cancer. An overview. Cancer Res. 44:5849–5958.

    Google Scholar 

  • MASUI, T., LECHNER, J.F. YOAKUM, G.H., WILEY, J.C. and HARRIS, C.C. Growth and differentiation of normal and transformed human bronchial epithelial cells. J. Cell Physiol. (Suppl) 4:73–81.

  • MASUI, T., WAKEFIELD, L.M., LECHNER, J.F., LA VECK, M.A., SPORN, M.B. and HARRIS, C.C. (1986). Type β transforming growth factor is the primary differentiation-inducing serum factor for normal human bronchial epithelial cells. Proc. Natl. Acad. Sci. USA 83:2438–2442.

    Google Scholar 

  • McDOWELL, E.M., BARRETT, L.A., GLAVIN, F., HARRIS, C.C. and TRUMP, B.F.(1978). The respiratory epithelium. I. Human bronchus. J. Natl. Cancer Inst. 61: 539–549.

    Google Scholar 

  • MILLER, B.E., MILLER, F.R., WILBURN, D. and HEPPNE, G.H. (1988). Dominance of a tumor subpopulation in mixed heterogeneous mouse mammary tumors. Cancer Res. 48:5747–5753.

    Google Scholar 

  • MIOH, H. and CHEN, J.-K. (1989). Transforming growth factor-β inhibits cellular adenylate cyclase activity in cultured human arterial endothelial cells. In Vitro Cell Dev. Biol. 25:101–104.

    Google Scholar 

  • MüLLER, K.M. and MüLLER, G. (1983). The ultrastructure of preneoplastic changes in the bronchial mucosa. In: Pulmonary Diseases: Clinical pathological Correlations, pp. 233–263. Springer-Verlag, Berlin.

    Google Scholar 

  • MUSTOE, T.A., PIERCE, G.F., THOMASON, A., GIAMATES, P., SPORN, M.B. and DEUEL, T.F. (1987). Accelerated healing of incisional wounds in rats induced by transforming growth factor-β. Science 237:1333–1336.

    Google Scholar 

  • NEEDLEMAN, B.W., CHOI, J., BURROWS-MEZU, A. and FONTANA, J.A. (1990). Secretion and binding of transforming growth factor beta by normal fibroblasts in scleroderma. Arthritis and Rheumatism 33:650–656.

    Google Scholar 

  • PERACCHIA, C. and BERNARDINI, G. (1984). Gap junction structure and cell-cell coupling regulation: Is there a calmodulin involvement? Fed. Proc. 43:2681–2691.

    Google Scholar 

  • PIERCE, G.B. and WALLACE, C. (1971). Differentiation of malignant to benign cells. Cancer Res. 31:127–134.

    Google Scholar 

  • REDDELL, R.R., KE, Y., GERWIN, B.I., McMENAMIN, M.G., LECHNER, J.F., SU, R.T., BRASH, D.E., PARK, J.-B., RHIM, J.S. and HARRIS, C.C. (1988). Transformation of human bronchial epithelial cells by infection with SV40 or adenovirus-12 SV 40 hybrid virus, or transfection via strontium phosphate coprecipitation with a plasmid containing SV40 early region genes. Cancer Res. 48:1904–1909.

    Google Scholar 

  • RESAU, J.H., ELLIGET, K.E., COTTRELL, J.R. and HUDSON, E.A. (1987). Cell injury and regeneration of human epithelium in organ culture. Cell Biol. Toxicol. 3:441–458.

    Google Scholar 

  • ROBERTS, A.B., ANZANO, M.A., WAKEFIELD, L.M., ROCHE, N.S., STERN, D.F. and SPORN, M.B. (1985). Type β transforming growth factor: A bifunctional regulator of cellular growth. Proc. Natl. Acad. Sci. USA 82:119–123.

    Google Scholar 

  • SACCOMANNO, G., ARCHER, V.E., AUERBACH, O., SAUNDERS, R.P. and BRENNAN, L.M. (1974). Development of carcinoma of the lung as reflected in exfoliated cells. Cancer 32:256–270.

    Google Scholar 

  • SCHULTZ, G.S., WHITE, M., MITCHELL, R., BROWN, G., LYNCH, J., TWARDZIK, D.R. and TODARO, G.J. (1987). Epithelial wound healing enhanced by transforming growth factor-alpha and vaccinia growth factor. Science 235:350–352.

    Google Scholar 

  • SHIROEDA, O., YAMAGUCHI, N. and KAWAI, K. (1987). Stimulation of low density lipoprotein receptor activity by conditioned medium from a human cancer cell line. Cancer Res. 47:4630–4633.

    Google Scholar 

  • SPORN, M.B., ROBERTS, A.B., WAKEFIELD, L.M. and ASSOIAN, R.K. (1980). Transforming growth factor-gb. Biological functional and chemical structure. Science 233:532–534.

    Google Scholar 

  • SPORN, M.B. and TODARO, G.J. (1980). Autocrine secretion and malignant transformation of cells. N. Engl. J. Med. 303:878–880.

    Google Scholar 

  • TEETER, S.M., HOLMES, F.F. and McFARLANE, M.J. (1987). Lung carcinoma in the elderly population: Influence of histology on the inverse relationship of stage to age. Cancer 60:1331–1336.

    Google Scholar 

  • TRUMP, B.F. and BEREZESKY, I.K. (1987). The role of ion regulation in respiratory carcinogenesis. In: Lung Carcinomas (E.M. McDowell, ed.), pp. 162–174. Churchill Livingstone, Inc., New York.

    Google Scholar 

  • TRUMP, B.F., RESAU, J. and BARRETT, L.A. (1980). Methods of organ culture for human bronchus. Methods of Cell Biology 21A:1–14.

    Google Scholar 

  • VAN DER MEEREN, A., LEVY, F., BIGNON, J. and JAURAND, M-C. (1988). Growth of normal and neoplastic pleural mesothelial cells in the presence of conditioned medium from neoplastic mesothelial cells. Biol. Cell 62:293–296.

    Google Scholar 

  • VAN DUREN, B.L. and GOLDSCHMIDT, B.M. (1976). Cocarcinogenic and tumor-promoting agents in tobacco carcinogenesis. J. Natl. Cancer Inst. 56:1237–1242.

    Google Scholar 

  • VENA, J.E., BYERS, T.E., COOKPAIR, D. and SWANSON, M. (1985). Occupation and lung cancer risk. An analysis by histologic subtypes. Cancer 56:910–917.

    Google Scholar 

  • WILLEY, J.C., MOSER, C.E. Jr., LECHNER, J.F. and HARRIS, C.C. (1984). Differential effects of 12-O-tetradecanoylphorbol-13-acetate on cultured normal and neoplastic human bronchial epithelial cells. Cancer Res. 44:5124–5126.

    Google Scholar 

  • WU, R., NOLAN, E. and TURNER, C. (1985). Expression of tracheal differentiated functions in serum-free, hormone-supplemented medium. J. Cell Physiol. 125:167–181.

    Google Scholar 

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Authors and Affiliations

  1. Department of Pathology, Human Tissue Resource, University of Maryland School of Medicine, 22 South Greene Street, 21201, Baltimore, Maryland

    Craig D. Albright, Raymond T. Jones, Eric A. Hudson, Benjamin F. Trump & James H. Resau

  2. University of Maryland Cancer Center, University of Maryland School of Medicine, 22 South Greene Street, 21201, Baltimore, Maryland

    Joseph A. Fontana

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  1. Craig D. Albright
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  2. Raymond T. Jones
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  3. Eric A. Hudson
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  4. Joseph A. Fontana
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  5. Benjamin F. Trump
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  6. James H. Resau
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Contribution No. 2801 from the Pathobiology Laboratory, University of Maryland.

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Albright, C.D., Jones, R.T., Hudson, E.A. et al. Transformed human bronchial epithelial cells (beas-2b) alter the growth and morphology of normal human bronchial epithelial cells in vitro . Cell Biol Toxicol 6, 379–398 (1990). https://doi.org/10.1007/BF00120804

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