alpha-chymotrypsin and Cell-Transformation--Neoplastic

The purpose of the studies described in this mini review article was to identify nontoxic compounds that could prevent or suppress the radiation induced malignant transformation of cells and be useful as human cancer preventive agents.. (1) Many different types of potential anticarcinogenic substances were evaluated initially for their abilities to prevent or suppress radiation induced malignant transformation in vitro, and certain anticarcinogenic protease inhibitors (APIs) were observed to be the most powerful anticarcinogenic agents at suppressing this surrogate endpoint biomarker of radiation carcinogenesis. (2) Within the category of APIs, those that inhibited the activity of chymotrypsin were effective at far lower molar concentrations than other APIs. The soybean-derived protease inhibitor known as the Bowman-Birk inhibitor (BBI) is a particularly powerful chymotrypsin inhibitor that is able to prevent radiation induced transformation in vitro (at concentrations down to nanomolar levels) as well as radiation induced carcinogenesis in vivo without toxicity. (3) There were many other unusual characteristics of APIs that led to the selection of one of these APIs, BBI, as the most appropriate compound for us to develop as a human cancer preventive agent. As one example, the APIs have an irreversible effect on carcinogenesis, while the effects are reversible for most anticarcinogenic agents when they are removed from carcinogenesis assay systems. (4) Numerous studies were performed in attempts to determine the potential mechanisms by which the APIs could prevent or suppress radiation induced carcinogenesis in in vitro and in vivo systems, and the results of these studies are described in this review article. The APIs and the proteases which interact with them appear to play important roles in radiation carcinogenesis. (5) Preparations for human trials using BBI began decades ago. The cost of preparing purified BBI was far too high to consider performing human trials with this agent, so BBI Concentrate (BBIC), a soybean extract enriched in BBI, was developed for the specific purpose of performing human trials with BBI. BBIC achieved Investigational New Drug (IND) Status with the Food and Drug Administration in April,1992, and human BBIC trials began at that time. (6) Several human trials were performed using BBIC and they indicated many potentially beneficial health effects produced by BBIC administration to people in various forms (e.g. tablets). 7) It is hypothesized that BBI takes the place of α-1-antichymotrypsin, an important regulatory compound in the human body, and helps to maintain homeostasis.

Topics: Anticarcinogenic Agents; Cell Transformation, Neoplastic; Chymotrypsin; Humans; Peptide Hydrolases; Protease Inhibitors; Trypsin Inhibitor, Bowman-Birk Soybean

In this report, we have investigated whether a protease inhibitor obtained from potatoes (chymotrypsin inhibitor 1; CI-1) will inhibit carcinogen-induced transformation of C3H/10T1/2 cells. CI-1 was as effective as the soybean-derived Bowman Birk inhibitor at suppressing radiation-induced transformation of C3H/10T1/2 cells, at a concentration of 10 micrograms/ml. The inhibitor was not toxic to the cells at concentrations of 0.1-10 micrograms/ml, the concentrations of CI-1 employed in the transformation experiments. To investigate the interaction of this inhibitor with the target cells, binding studies were carried out. 125I-labelled CI-1 could not be displaced from C3H/10T1/2 cells by co-incubation of the cells with a 5000-fold excess of unlabelled inhibitor. These results suggest that this inhibitor does not reversibly bind to specific receptor proteins on the surface of these cells.

Topics: Animals; Cell Transformation, Neoplastic; Cells, Cultured; Chymotrypsin; Mice; Mice, Inbred C3H; Plant Proteins; Solanum tuberosum; X-Rays

Topics: Animals; Carcinogens; Cell Transformation, Neoplastic; Chymotrypsin; DNA Damage; Glycine max; Humans; Mice; Pancreatic Neoplasms; Protease Inhibitors; Skin Neoplasms; Superoxides; Trypsin Inhibitors

Experiments reported here indicate a crude soybean extract, if defatted with acetone, effectively blocks cell transformation in vitro. An active component of this crude extract is the Bowman-Birk trypsin and chymotrypsin inhibitor. The chymotrypsin-inhibitory region of the Bowman-Birk inhibitor is responsible for suppressing in vitro transformation. Another low molecular weight soybean trypsin inhibitor does not significantly suppress transformation. The Bowman-Birk inhibitor (i) has an irreversible effect on the transformation process, (ii) can suppress radiation-induced transformation even when added to cultures many days after the carcinogen exposure, and (iii) is effective in its ability to suppress transformation when present in the medium at a concentration as low as 0.125 nM.

Topics: Acetone; Animals; Cell Transformation, Neoplastic; Cells, Cultured; Chymotrypsin; Mice; Mice, Inbred C3H; Trypsin Inhibitor, Bowman-Birk Soybean; Trypsin Inhibitors; X-Rays

Alpha-1-antitrypsin (A1AT) has been shown to be a useful tumor marker for hepatocellular carcinoma (HCC) and some other neoplasms. No studies of alpha-1-antichymotrypsin (A1AChy) in HCC have been reported. A comparative study of A1AT and A1AChy in HCC was undertaken. While 19/33 HCC were positive for A1AT, all 33 HCC contained immunoreactive A1AChy. Cells showing positive immunoreaction for both A1AT and A1AChy were more numerous in moderately differentiated HCC than in well differentiated or poorly differentiated HCC. Although the pattern of staining with both antisera was similar, in cases showing positive staining for both antisera, A1AChy-positive cells were more numerous than A1AT-positive cells. The results suggest a useful role for A1AChy as a marker of HCC.

Topics: alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; Carcinoma, Hepatocellular; Cell Transformation, Neoplastic; Chymotrypsin; Histocytochemistry; Humans; Hyperplasia; Immunoenzyme Techniques; Liver; Liver Cirrhosis; Liver Neoplasms

Normal and transformed fibroblasts can be discriminated by a flow cytometry assay on the basis of their differential reaction with fluorescamine. The cause of altered reactivity of transformed cells with this fluorescent probe has been investigated by a detailed analysis of its reaction with chicken embryo fibroblasts transformed by a temperature sensitive mutant of Rous sarcoma virus. The subcellular distribution of fluorescent adducts characterized by cell fractionation and gel electrophoresis procedures supports the hypothesis that transformed cells possess a surface barrier which decreases the accessibility of fluorescamine to reactive macromolecules. The barrier has been identified as being composed at least partly of hyaluronic acid, because of the ability of purified and specific hyaluronidase (from Streptomyces hyalurolyticus) to modulate the response of transformed cells to fluorescamine. Enzyme treatment of transformed cells prior to reaction with fluorescamine causes them to resemble nontransformed cells both in the nature of components labeled and in their fluorescence intensity. It is suggested that fluorescamine monitors an altered surface hyaluronic acid composition which occurs upon transformation. Its significance is discussed in terms of the known physical properties of the molecule and the finding that it is an early event in the process of transformation.

Topics: Animals; Avian Sarcoma Viruses; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Chick Embryo; Chymotrypsin; Electrophoresis, Polyacrylamide Gel; Fibroblasts; Flow Cytometry; Fluorescamine; Hyaluronic Acid; Hyaluronoglucosaminidase; Spiro Compounds

EGTA dissociated cell lines could be distinguished on the basis of their dependence on calcium during initial aggregation. However, the calcium-dependent aggregation could not be abolished by low chymotrypsin/EGTA treatment. These differences were not related to any morphogenetic property of the cells and suggest that different aggregation mechanisms are involved.

Topics: Animals; Calcium; Cell Adhesion; Cell Aggregation; Cell Line; Cell Membrane; Cell Transformation, Neoplastic; Chymotrypsin; Dogs; Egtazic Acid; Ethylene Glycols; Humans

Topics: Animals; Cathepsins; Cell Transformation, Neoplastic; Chemical Phenomena; Chemistry; Chromatography, Gel; Chymotrypsin; Hydrogen-Ion Concentration; Leukocytes; Neoplasm Metastasis; Pancreatic Elastase; Protease Inhibitors; Swine

BALB/c mouse 3T3 cells transformed by simian virus 40 (SV3T3), baby hamster kidney cells transformed by polyoma virus or Rous sarcoma virus, and a range of neoplastic human cell lines release material that inhibits the migration of macrophages and lymphocytes. Similar migration-inhibitory factor (MIF) activity was not detected in supernatants from cultures of untransformed 3T3 or baby hamster kidney cells and a variety of human diploid cell strains. Physico-chemical characterization of the MIF produced by SV3T3 and HeLa cells revealed substantial similarities with the MIF produced by mitogen-activated human peripheral lymphocytes. MIF released by tumor cells is inhibited by pancreatic and soybean trypsin inhibitors and by diisopropylfluorophosphate, indicating that it is a serine-protease. Comparison of MIF produced by SV3T3 cells with a serine-protease plasminogen activator released by the same cells indicated that the latter is more heat labile and has a more heterogenous elution profile after chromatography on Sephadex G-75. The possible role of MIF in causing proteolytic modification of the surface properties of tumor cells and in altering cell-mediated immune responses to neoplastic cells is discussed.

Topics: Animals; Avian Sarcoma Viruses; Cell Line; Cell Migration Inhibition; Cell Transformation, Neoplastic; Cell-Free System; Chymotrypsin; Cricetinae; Endopeptidases; HeLa Cells; Hot Temperature; Humans; Isoflurophate; Lymphocytes; Macrophage Migration-Inhibitory Factors; Macrophages; Mice; Mice, Inbred BALB C; Neoplasms; Polyomavirus; Serine; Simian virus 40; Trypsin Inhibitors

Topics: Amino Acid Sequence; Aminocaproates; Antibodies; Antigen-Antibody Complex; Antigen-Antibody Reactions; Antigens, Neoplasm; Antilymphocyte Serum; Cell Membrane; Cell Transformation, Neoplastic; Chymotrypsin; Complement System Proteins; Epitopes; Esterases; Humans; Immunoglobulin G; Lectins; Leukocidins; Lymphocyte Activation; Papain; Peptides; Staphylococcus; Streptolysins; T-Lymphocytes; Toxins, Biological; Trypsin

Topics: Animals; Carbon Radioisotopes; Cell Line; Cell Transformation, Neoplastic; Centrifugation, Density Gradient; Chymotrypsin; Electrophoresis, Polyacrylamide Gel; Fibroblasts; Gammaretrovirus; Glucosamine; Glycoproteins; Iodine Radioisotopes; Leucine; Molecular Weight; Moloney murine leukemia virus; Peroxidases; Rats; Rats, Inbred Strains; Sarcoma; Tritium; Trypsin; Viral Proteins

Topics: Agglutination Tests; Animals; Binding Sites, Antibody; Cell Membrane; Cell Transformation, Neoplastic; Cells, Cultured; Chymotrypsin; Glutathione; Ketones; Lectins; Leukemia L1210; Mice; Mitosis; Peptide Hydrolases; Tosyl Compounds

Topics: Animals; Benz(a)Anthracenes; Cell Division; Cell Nucleus; Cell Transformation, Neoplastic; Chymotrypsin; DNA; Epithelial Cells; Epithelium; Fluorometry; Hyperplasia; Male; Methods; Mice; Mitosis; Neoplasms, Experimental; Precancerous Conditions; Skin; Skin Neoplasms

Three synthetic inhibitors of proteases (tosyl lysine chloromethyl ketone, tosyl phenylalanine chloromethyl ketone, and tosyl arginine methyl ester) inhibit the tumorigenesis initiated in mouse skin by 7,12-dimethylbenz(a)anthracene and promoted by croton oil or its active principle, phorbol ester. These protease inhibitors, when applied directly to mouse skin, inhibit some of the irritant effects of the tumor promoter and are not toxic.

Topics: Animals; Antineoplastic Agents; Arginine; Benz(a)Anthracenes; Cell Transformation, Neoplastic; Chymotrypsin; Croton Oil; Depression, Chemical; Esters; Ketones; Lysine; Mice; Papain; Phenylalanine; Skin Neoplasms; Terpenes; Trypsin Inhibitors