Growth Inhibition and Morphological Changes Caused by Lipophilic Acids in Mammalian Cells

Abstract

Human (BIeLa, Chang liver, L-132, and Intestine 407) and other mammalian (XC, SV3T3, and chick-embryo) cells in tissue culture are at least as sensitive to inhibition by lipophilic acids and nitrite as bacteria. Some of these compounds are the most frequently used antimicrobial food additives. Short-chain fatty acids (up to hexanoate) and parabens induce, at partially inhibitory concentrations, a jagged cell shape in continuous epithelial-like cell lines, such as HeLa, Chang liver, L-132, and Intestine 407. This morphological effect is not mediated or enhanced by butyryl cyclic AMP, which specifically affects fibroblasts.Fatty acids and other lipophilic acids, some of which are used as antimicrobial food additives, inhibit the growth of Bacillus subtilis and the uptake of amino and keto acids into bacterial membrane vesicles (1-3). Since these results indicated that lipophilic acids alter the properties of cell membranes, it appeared likely that they would also affect mammalian cells. This report demonstrates that they inhibit the growth of mammalian cells at least as effectively as that of bacteria. In addition, it shows that short-chain lipophilic acids induce filamentous protrusions in epithelial-like cells but not in cells with fibroblast morphology. These morphological changes are not mediated or enhanced by butyryl cyclic AMP (BucAMP), which alters the morphology of fibroblasts but not of epithelial cells (4, 5). MATERIALS AND METHODSCell Cultures. HeLa cells, strain R, were obtained from Grand Island Biological Co. (GIBCO), Grand Island, N.Y. and propagated in our laboratory. Cells of Chang liver (normal human epithelial cells), L-132 (originating from human embryonic lung), and Intestine 407 (epithelial cells originally derived from human embryonic jejunum and ileum) were bought from Flow Laboratories, Rockville, Md. XC cells (a cell line derived from a rat sarcoma induced by Rous sarcoma virus) were a gift from Dr. Janet Hartley, National Cancer Institute, Bethesda, Md. Chick-embryo cells were prepared from 9-day-old chick embryo (6). All the above cells were cultivated in 60-mm plastic petri dishes or 75-cm2 T flasks (Falcon Plastics, Los Angeles, Calif.) in Eagle's minimal essential medium (pH 7.4) containing 10% fetal-calf serum (GIBCO), 2 mM L-glutamine, 100 units/ml of penicillin, and 100 /Ag/ml of kanamycin. The medium containing all these additions will in the following be called "minimal essential medium." The cultures were grown at 370 in a humidified incubator provided with 4% CO2. Unless specified otherwise, HeLa cell cultures in 60-mm petri dishes were in- Chemicals. Stock solutions of 100 mM Na propionate and Na butyrate were prepared in minimal essential medium, while 2.5 M Na hexanoate (pH 7.0) was prepared in distilled water. 1 M solution of 'octanoic and decanoic acid, methyl-and propyl-paraben (paraben = p-hydroxybenzoic acid ester) were prepared in 99% dimethyl sulfoxide (Matheson, Coleman &-Bell, spectral grade) and diluted to the desired concentrations in minimal essential m...

Characterization of BCE-1, a Transcriptional Enhancer Regulated by Prolactin and Extracellular Matrix and Modulated by the State of Histone Acetylation

“…The constructs were stably transfected into CID-9 cells, and the activity of each was com-VOL. 18,1998 ECM-RESPONSIVE ELEMENT AND GENE EXPRESSION 2185…”

“…6A and data not shown). Since sodium butyrate treatment has been shown to induce many changes in the cell in addition to the inhibition of histone deacetylase (18,24,27, 40), we tested a more specific inhibitor of histone deacetylase, trichostatin A (62). A dose-dependent induction of transcription of up to 10-fold was observed in cells cultured with trichostatin A in the absence of ECM (Fig.…”

Histone deacetylases and cancer

“…A defect in any of these processes may result in cancer, or the uncontrolled growth of the cell. It has been appreciated for decades that histone deacetylases (HDACs) play an important role in cancer development since the first observation that sodium butyrate (NaB), later identified as an HDAC inhibitor, can cause the morphological reversion of the transformed cell phenotype (Ginsburg et al, 1973;Altenburg et al, 1976;Boffa et al, 1978). Currently, HDAC inhibitors are in clinical trials to treat a variety of malignancies.…”

Butyrate does not induce single strand breaks in friend erythroleukemia cells or in 3T6 mouse fibroblasts

“…It induces terminal differentiation in some cell systerm [1][2][3][4], inhibits the synthesis of differentiationspecific proteins in others [5 ] and generally causes a block in DNA replication and cell division [6][7][8][9]. Cells grown in the presence of butyrate accumulate acetylated forms of histones (particularly of histones H3 and H4) due to an inhibition ofhistone deacetylase [10][11][12][13].…”