This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Willey, J. C. Articles by Harris, C. C. Search for Related Content PubMed PubMed Citation Articles by Willey, J. C. Articles by Harris, C. C.

Biochemical and Morphological Effects of Cigarette Smoke Condensate and Its Fractions on Normal Human Bronchial Epithelial Cells in Vitro

Laboratory of Human Carcinogenesis, National Cancer Institute, NIH, Bethesda, Maryland 20892 [J. C. W., C. E. M., C. O., C. C. H.]; and Department of Toxicology, Karolinska Institute, S-10401 Stockholm, Sweden [R. C. G., K. S.]

We investigated the effect of cigarette smoke condensate (CSC), two basic fractions (B_Ia and B_Ib) of CSC, the ethanol-extracted weakly acidic fraction (WA_e), and the methanol-extracted neutral fraction (N_meoh) on the clonal growth rate, plasminogen activator (PA) activity, cross-linked envelope (CLE) formation, and ornithine decarboxylase activity, epidermal growth factor (EGF) binding, thiol levels, and DNA single strand breaks in cultured human bronchial cells. Neither CSC nor any of the fractions were mitogenic over the range 0.01–100 µg/ml. All were growth inhibitory at higher concentrations. The 40% growth inhibitory concentrations for CSC, B_Ia, B_Ib, WA_e, and N_meoh were 10, 10, 10, 3, and 1 µg/ml, respectively. Effects on CLE formation, morphology, PA, and ornithine decarboxylase activities, EGF binding, and thiol levels were evaluated using 40% growth inhibitory concentrations. We found that CSC and all fractions caused an increased formation of CLEs, from a baseline of 0.5% in the untreated cells to a maximum increase of 25% induced by N_meoh. A squamous morphological change was observed within 1 h after exposure to N_meoh, WA_e, and CSC. The B_Ia and B_Ib fractions had little effect. Only N_meoh increased PA significantly, from 2.5 ± 0.4 to 5.1 ± 0.3 units/mg cellular protein. CSC and the WA_e and N_meoh (N_meoh > WA_e > CSC) fractions caused a decrease in EGF binding, in each case reaching a maximum effect after a 10–12-h incubation. This effect on EGF binding was further characterized in the case of N_meoh. In untreated normal human bronchial epithelial cells, by Scatchard analysis the k_d was 2.0 nM and there were 1.2 x 10⁵ receptors/cell. In cells incubated in medium containing N_meoh (3 µg/ml) the k_d was 3.2 nM and there were 1.1 x 10⁵ receptors/cell. Thus, inhibition of EGF binding by N_meoh was due primarily to a decrease in the affinity. At the 40% growth inhibitory concentrations neither CSC nor any of the fractions significantly affected intracellular thiol levels. While a 3-h incubation in medium containing CSC caused significant DNA single strand breaks only at a concentration of 100 µg/ml, N_meoh caused a marked effect at 5 µg/ml. Neither CSC nor any of the fractions had an effect on ornithine decarboxylase activity. Due to the effects of the N_meoh fraction on growth, morphology, EGF binding, PA activity, and formation of single strand breaks we consider it to be the most likely portion of CSC to contain compounds with actions similar to those of the phorbol ester, indole alkaloid, and polyacetate tumor promoters.

¹ Supported in part by the Swedish Cancer Society, the Swedish National Board for Laboratory Animals, the Workers Environment Health Fund, and the Swedish Tobacco Company.

² To whom requests for reprints should be addressed, at Building 37, Room 2C09, National Cancer Institute, NIH, Bethesda, Maryland 20892.

Received 7/22/86. Revised 6/10/86. Revised 1/13/87. Accepted 1/16/87.

This article has been cited by other articles:

				W. R. Fields, R. M. Leonard, P. S. Odom, B. K. Nordskog, M. W. Ogden, and D. J. Doolittle Gene Expression in Normal Human Bronchial Epithelial (NHBE) Cells Following In Vitro Exposure to Cigarette Smoke Condensate Toxicol. Sci., July 1, 2005; 86(1): 84 - 91. [Abstract] [Full Text] [PDF]

				A. Arora, C. A. Willhite, and D. C. Liebler Interactions of {beta}-carotene and cigarette smoke in human bronchial epithelial cells Carcinogenesis, August 1, 2001; 22(8): 1173 - 1178. [Abstract] [Full Text] [PDF]