bromochloroacetic-acid and cobaltous-chloride

Invasion is a hallmark of malignancy. The aim of this study was to develop an in vitro model that can be used for experimental studies of cancer cell invasion. The organotypic oral cancer model was constructed by growing oral squamous cell carcinoma (OSCC) cells on a collagen matrix in which normal human fibroblasts were incorporated. Immunohistochemical staining of the model showed that the expression of invasion-related molecules such as phosphorylated extracellular signal-regulated kinases 1 and 2 (p-ERK1/2), cyclooxygenase-2 (COX-2), p75(NTR), and hepatocyte growth factor receptor (Met) was similar to that seen in OSCC. Treatment of the model with cobalt chloride (CoCl(2)) to mimic hypoxic conditions increased cancer cell invasion, defined as the appearance of cancer cell islands protruding into the matrix. Models treated with CoCl(2) showed increased expression of p75(NTR) and laminin-5 in the cancer cells, and a more pronounced fragmentation of collagen IV in the basal membrane area, in contrast to models that were left untreated. The results indicate that the present model is well suited for studies on cancer cell invasion in the matrix and that the addition of CoCl(2) on day 3 of the experiment is indicated because it markedly increases the invasion and improves the model.

Topics: Antigens, CD; Carcinoma, Squamous Cell; Cell Culture Techniques; Cells, Cultured; Cobalt; Collagen; Collagen Type IV; Culture Media; Cyclooxygenase 2; Fibroblasts; Humans; Hypoxia; Keratins; Laminin; Male; Middle Aged; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Neoplasm Invasiveness; Nerve Tissue Proteins; Proto-Oncogene Proteins c-met; Receptors, Growth Factor; Receptors, Nerve Growth Factor; Tongue Neoplasms; Young Adult

Cultured cell lines are routinely used for in vitro toxicity screens, reducing the requirement for animal studies during the development of new pharmaceutical, agrochemical and cosmetic products. The foetal rat lung epithelial (FRLE) cell line was originally derived from alveolar type II cells (ATII) of the lung. The aims of this study were to further characterise FRLE cells and investigate their potential for screening for pneumotoxins. The cells were found to have retained some of the features of their progenitor cells, namely the expression of cytokeratin proteins, specifically cytokeratin 18, and the ability to actively accumulate the non-selective contact herbicide paraquat. However, the cells have lost the ability to synthesise surfactant protein mRNA and no longer contain multiple lamellar bodies. Toxins that damage ATII cells in vivo (cadmium chloride, cobalt chloride and paraquat) were found to induce cytotoxicity in FRLE cells, as did the non-specific pneumotoxin nitrofurantoin, and hydrogen peroxide. However, the cells were less sensitive to the effects of compounds that require metabolic activation (1-nitronaphthalene, coumarin and butylated hydroxytoluene) and the hepatotoxin bromobenzene. Thus, FRLE cells appear to be a good in vitro model for monitoring the potential toxicity to ATII cells and could be used as an initial screen for pneumotoxicity.

Topics: Animals; Bromobenzenes; Cadmium Chloride; Cell Line; Cobalt; Cytotoxins; Drug Evaluation, Preclinical; Epithelial Cells; Fetus; Gene Expression; Humans; Hydrogen Peroxide; In Situ Hybridization; Keratins; Lung; Mice; Microsomes, Liver; Naphthalenes; Neutral Red; Nitro Compounds; Nitrofurantoin; Paraquat; Pulmonary Alveoli; Pulmonary Surfactant-Associated Protein A; Pulmonary Surfactant-Associated Protein B; Rats; RNA, Messenger