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Effects of Serum, Transforming Growth Factor Type ß, or 12-O-Tetradecanoylphorbol-13-acetate on Ionized Cytosolic Calcium Concentration in Normal and Transformed Human Bronchial Epithelial Cells

Laboratory of Human Carcinogenesis, Division of Cancer Etiology, National Cancer Institute, Bethesda, Maryland 20892 [M. M., J. C. W., J. F. L., C. C. H.], and Department of Pathology, University of Maryland School of Medicine, Baltimore, Maryland 21201 [M. W. S., B. F. T.]

Fluctuations in ionized cytosolic calcium ([Ca²⁺]_i) are considered important signals for induction of growth or differentiation in mammalian cells. The resting concentrations of [Ca²⁺]_i in normal and adenovirus 12-SV40 hybrid virus-transformed (BEAS-2B) human bronchial epithelial cells were 63 ± 15 nM (SD) and 44 ± 15 nM, respectively. Eight % calcium-free fetal bovine serum rapidly caused a significant increase in [Ca²⁺]_i, while causing both cell types to undergo squamous differentiation. When treated with 8% calcium-free fetal bovine serum, a serum-sensitive subclone of BEAS-2B cells exhibited a higher elevation of [Ca²⁺]_i than a serum-resistant (i.e., not stimulated to differentiate by serum) subclone. However, a serum component involved in the induction of squamous differentiation, transforming growth factor type ß, did not increase [Ca²⁺]_i in either normal cells or BEAS-2B cells. 12-O-Tetradecanoylphorbol-13-acetate, an exogenous inducer of squamous differentiation and activator of protein kinase C, did not increase [Ca²⁺]_i, but did attenuate serum-induced elevation of [Ca²⁺]_i. These results suggest that while an increase in [Ca²⁺]_i is associated with serum-induced squamous differentiation, a cytosolic ionized calcium signal is not required for the initiation of the squamous differentiation pathway induced by either transforming growth factor type ß or 12-O-tetradecanoylphorbol-13-acetate.

¹ To whom requests for reprints should be addressed, at Building 37, Room 2C01, National Cancer Institute, Bethesda, MD 20892.

Received 2/23/88. Revised 8/18/88. Accepted 10/ 5/88.

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