sodium-dodecyl-sulfate and Carcinoma--Squamous-Cell

It is important to detect mediastinal lymph node metastases in patients with lung cancer to improve outcomes, and it is possible that activatable fluorescence imaging with indocyanine green (ICG) can help visualize metastatic lymph nodes. Therefore, we investigated the feasibility of applying this method to mediastinal lymph node metastases in an epidermal growth factor receptor (EGFR)-positive squamous cell carcinoma of the lung. Tumors were formed by injecting H226 (EGFR-positive) and H520 (EGFR-negative) cell lines directly in the lung parenchyma of five mice each. When computed tomography revealed tumors exceeding 8 mm at their longest or atelectasis that occupied more than half of lateral lung fields, a panitumumab (Pan)-ICG conjugate was injected in the tail vein (50 μg/100 μL). The mice were then sacrificed 48 hours after injection and their chests were opened for fluorescent imaging acquisition. Lymph node metastases with the five highest fluorescent signal intensities per mouse were chosen for statistical analysis of the average signal ratios against the liver. Regarding the quenching capacity, the Pan-ICG conjugate had almost no fluorescence in phosphate-buffered saline, but there was an approximate 61.8-fold increase in vitro after treatment with 1% sodium dodecyl sulfate. Both the fluorescent microscopy and the flow cytometry showed specific binding between the conjugate and H226, but almost no specific binding with H520. The EGFR-positive mediastinal lymph node metastases showed significantly higher average fluorescence signal ratios than the EGFR-negative ones (n = 25 per group) 48 hours after conjugate administration (70.1% ± 4.5% vs. 13.3% ± 1.8%; p < 0.05). Thus, activatable fluorescence imaging using the Pan-ICG conjugate detected EGFR-positive mediastinal lymph node metastases with high specificity.

Topics: Animals; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Line, Tumor; ErbB Receptors; Female; Fluorescent Dyes; Heterografts; Humans; Indocyanine Green; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Mediastinum; Mice; Mice, Nude; Microscopy, Fluorescence; Optical Imaging; Panitumumab; Photochemistry; Sodium Dodecyl Sulfate; Tomography, X-Ray Computed

Toothpastes contain three main components: detergents, abrasives, and fluoride. Detergents, particularly sodium lauryl sulfate, have been proposed as components that enable toothpastes to produce cytotoxic effects in vitro. However, not all toothpastes contain sodium lauryl sulfate, and almost no studies have found an association between detergents and the in vitro cytotoxicity of toothpastes. The present study examined the in vitro cytotoxicity of nine commercially available toothpastes containing four different detergents. Toothpastes were diluted in serum-free medium, centrifuged, and filter sterilized. The half-lethal concentration of the toothpaste-conditioned medium (TCM) was calculated based on the formation of formazan by gingival fibroblasts, oral squamous cell carcinoma HSC-2 cells, and L929 cells. Cell proliferation was analyzed, and live-dead staining was performed, after exposure of cells to conditioned medium prepared with 1% toothpaste (1% TCM). It was found that toothpastes containing sodium lauryl sulfate and amine fluoride strongly inhibited cell viability with the half-lethal concentration being obtained with conditioned medium prepared with approximately 1% toothpaste (1% TCM). Toothpastes containing cocamidopropyl betaine and Steareth-20 showed higher half-lethal concentration values, with the half-lethal concentration being obtained with conditioned medium prepared with 10% (10% TCM) and 70% (70% TCM) toothpaste, respectively. Proliferation and live-dead data were consistent with the cell-viability analyses. These results demonstrate that the type of detergent in toothpastes can be associated with changes in in vitro cell toxicity.

Topics: Animals; Betaine; Carcinoma, Squamous Cell; Cariostatic Agents; Cell Culture Techniques; Cell Line; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cells, Cultured; Culture Media, Conditioned; Culture Media, Serum-Free; Detergents; Diamines; Epithelial Cells; Fibroblasts; Fluorescent Dyes; Fluorides; Formazans; Gingiva; Humans; Indicators and Reagents; Materials Testing; Mice; Mouth Neoplasms; Polyethylene Glycols; Sodium Dodecyl Sulfate; Tetrazolium Salts; Toothpastes

Patients receiving radiation therapy due to oral cancer develop complications such as hyposalivation, mucositis, oral infections, dental hypersensitivity and caries. Mouthrinses can alleviate some of these problems.. To investigate the in vitro antimicrobial properties and cytotoxicity of an experimental mouthrinse.. The mouthrinse contained 30% hexylene glycol (glycerine), 7% potassium nitrate and 0.025% sodium fluoride. The minimal inhibitory concentration (MIC) of these ingredients and the mixture was determined for C. albicans, S. aureus and S. mutans over 24 hours at different concentrations. The MICs of two commercial mouthrinses, Corsodyl and Plax, were also determined using the same organisms. All mouthrinses were then tested to determine the percentage kill over 1, 2, and 3 minutes.. The MICs for hexylene glycol were 10%, 30% and 10% for C. albicans, S. aureus and S. mutons respectively. Potassium nitrate and sodium fluoride had no antimicrobial effects. The MIC of Corsodyl was 0.016 mg/ml for all the test organisms. The MIC for Plax varied from 0.0002 mg/ml to 0.001 mg/ml. The kill rates for all mouthrinses were acceptable, with no statistical differences between them. The experimental mouthrinse was not toxic to human oesophageal SCC cells after 1 minute exposure. At the time of the experiment, the costs of a similar quantity of the experimental mouthrinse, Corsodyl and Plax were R5.24, R30.00 and R10.00 respectively.. The experimental mouthrinse was cost-effective and proved to have an antimicrobial effect and could be used safely to alleviate oral infections, desensitize teeth, improve oral hygiene and control dental caries in cancer patients after radiation therapy.

Topics: Anti-Infective Agents, Local; Benzoates; Candida albicans; Carcinoma, Squamous Cell; Cariostatic Agents; Cell Adhesion; Cell Line, Tumor; Chlorhexidine; Dentin Desensitizing Agents; Dose-Response Relationship, Drug; Esophageal Neoplasms; Glycols; Humans; Lubricants; Materials Testing; Microbial Sensitivity Tests; Mouthwashes; Nitrates; Potassium Compounds; Radiotherapy; Sodium Dodecyl Sulfate; Sodium Fluoride; Staphylococcus aureus; Streptococcus mutans; Time Factors; Triclosan

The aim of this study was to investigate the heat stability of squamous cell carcinoma (SCC) antigen, a tumor-associated serine proteinase inhibitor (serpin), in tumor tissue extract by electrophoretic methods. After heat treatment at 70 degrees C for 2 h, the tumor tissue extract showed a single main protein band of 45 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) which reacted with a monoclonal antibody specific for SCC antigen. The heat-stable SCC antigen was separated by two-dimensional electrophoresis (2-DE) into four spots with pI 6.4-5.9 and Mr 44500-45 000 of SCC antigen-1. Furthermore, the SCC antigen-1 still showed its inhibitory activity against a cysteine proteinase, papain, by gelatin zymography. These results suggest that heat treatment of protein sample at 70 degrees C for 2 h may be a useful method for a partial purification of SCC antigen-1 which can inhibit lysosomal cysteine proteinases such as cathepsin L, S, and K.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Squamous Cell; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Heating; Humans; Serpins; Sodium Dodecyl Sulfate; Tissue Extracts

Guanidinobenzoatase (GB) is a cell surface proteolytic enzyme capable of degrading fibronectin, and is associated with tumour cells and cells capable of migration. The location of active GB in sections has been demonstrated with 9-aminoacridine (9-AA), a competitive inhibitor of GB. 3,4-Dichloroisocoumarin (3,4-DCI) and pentamidine isethionate (PI) are inhibitors of trypsin-like enzymes. It has now been demonstrated that 3,4-DCI, PI, and guanidino derivative compounds are significant inhibitors of GB, on the surfaces of lung squamous cell carcinoma cells in frozen sections and free GB in solution. Dexamethasone acetate (DMA) and medroxy-progesterone (MP) did not show any significant inhibition of GB activity. These molecules lack a reactive chloride or guanidino groups and are thought to react at the nuclear level, rather than directly on this cell surface protease. Kinetic studies have shown that 3,4-DCI, PI and guanidino derivatives are reversible competitive inhibitors of GB, as determined in vitro on the purified enzyme. The inhibition resulting with 3,4-DCI was a time-dependent process. It is suggested that these inhibitors interact with GB by binding to its active site, resulting in the formation of enzyme-inhibiter complexes (GB-I). The GB-I complexes can be dissociated with SDS treatment, resulting in the regain of GB activity.

Topics: Benzoates; Binding, Competitive; Carboxylic Ester Hydrolases; Carcinoma, Squamous Cell; Cell Membrane; Coumarins; Dexamethasone; Endopeptidases; Guanidines; Humans; Isocoumarins; Kinetics; Lung Neoplasms; Medroxyprogesterone; Microscopy, Fluorescence; Pentamidine; Protease Inhibitors; Sodium Dodecyl Sulfate; Sulfaguanidine

Matrix metalloproteinases (MMPs) are believed to play an important role in tumor invasion and metastasis. MMPs have been identified as proforms of malignant tumor-associated enzymes, such as procollagenase (proMMP-1) of M(r) = 53,000, progelatinase (proMMP-2) of M(r) = 72,000, proMMP-9 of M(r) = 92,000, and prostromelysin (proMMP-3) of M(r) = 59,000. Here we report that two cell lines of squamous cell carcinoma (SCC9 and SCC25) produce at least two matrix metalloproteinases (MMPs) in zymogen form, which have been identified as proMMP-2 and 3 by indirect immunofluorescence technique, immunoblot analysis, and gelatin-substrate gel enzymography. Additionally, a 92-kDa gelatinolytic metalloproteinase (proMMP-9) was detected by gelatin-substrate gel enzymography. We propose that the ability of these tumor cells to secrete MMPs plays an important role in the malignant behavior of oral squamous cell carcinomas.

Topics: Adult; Aged; Carcinoma, Squamous Cell; Collagenases; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Humans; Immunoblotting; Male; Matrix Metalloproteinase 3; Matrix Metalloproteinase 9; Metalloendopeptidases; Neoplasm Proteins; Sodium Dodecyl Sulfate; Tongue Neoplasms; Tumor Cells, Cultured

Urokinase-type (uPA) and tissue-type (tPA) plasminogen activators were identified by fibrinolytic autography in the sulcus epithelium of human gingival mucosa but not in the orthokeratinized gingival epithelium. Fibrinolytic activity was present only over blood vessels in frozen sections of oral squamous cell carcinomas, the malignant epithelial cells showing no plasminogen activator activity. Plasminogen activators could not be demonstrated in either the sulcus or gingival epithelium by immunofluorescence, but both uPA and tPA were found in occasional squamous carcinoma cells. Fibrinolytic activity of culture fluids from epithelial explants grown in vitro from human gingival mucosa showed marked variation, but activity was much higher in the culture supernatants than in the cell lysates. Fibrinolytic activity of culture fluids from epithelial explants of squamous cell carcinomas was low both in supernatants and lysates. Zymogram overlays of sodium dodecyl sulphate-polyacrylamide electrophoretic gels from culture supernatants showed that the low fibrinolytic activity of culture supernatants of oral squamous cell carcinomas was due to the associated presence of plasminogen activator inhibitors. The fibrinolytic activity in the zymogram was due predominantly to uPA but some lysis was due also to tPA.

Topics: Carcinoma, Squamous Cell; Cells, Cultured; Connective Tissue; Electrophoresis, Polyacrylamide Gel; Epithelial Cells; Epithelium; Fibrinolysis; Fluorescent Antibody Technique; Gingiva; Humans; In Vitro Techniques; Keratinocytes; Molecular Weight; Mouth Neoplasms; Sodium Dodecyl Sulfate; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator

Cytokeratin pattern was analyzed in 14 moderately differentiated and 12 well-differentiated squamous cell carcinomas of buccal mucosa by SDS-PAGE, immunoblotting and two dimensional electrophoresis. These were compared with patterns of normal buccal mucosa and surrounding areas whenever possible. Normal buccal mucosa expresses keratin No. 4 (59Kd), 5 (58Kd), 13 (54Kd) and 14 (50Kd). Keratin No. 4 (59Kd) and 14 (50Kd) were expressed by 20 of 26 tumors studied, while many of the tumors did not express keratins No. 5 (58Kd) and 13 (54Kd). Keratin No. 1 (67Kd) and 16 (48Kd) were aberrantly expressed by 9 well-differentiated tumors. Keratin No. 17 (46Kd) and 18 (45Kd) were expressed by 10 and 8 tumors of 14 moderately differentiated tumors. Six tumors which showed involvement of alveolar mucosa, expressed some keratins expressed by its normal counterpart. Their altered expression was consistent with the differentiation pattern as stated earlier. Non-expression of keratins 5 and 13 seems to be the result of malignant transformation and is seen in the majority of tumors, while appearance of aberrant keratins seems to be related more to the degree of differentiation of the tumor.

Topics: Carcinoma, Squamous Cell; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Humans; Immunoblotting; Isoelectric Focusing; Keratins; Mouth Mucosa; Mouth Neoplasms; Sodium Dodecyl Sulfate

The frequency of the occurrence of cytokeratins analyzed SDS-electrophoretically in 20 leukoplakic lesions and 14 squamous cell carcinomas of the oral mucosa has shown a picture of "restlessness" with some quantitative, some qualitative deviations from the locally normal pattern of cytokeratins. In most cases the basic pattern typical for the oral mucosa was still recognizable, except for three highly undifferentiated carcinomas. The variations of the cytokeratin pattern existed independently of the clinical form of leukoplakia and of its histological degree of dysplasia. The striking findings in some, but not all cases were: --the presence of cytokeratin no. 18 as a major component which normally appears rarely and faintly, and of cytokeratin no. 19, which is not normally detectable electrophoretically within the whole oral mucosa; --the absence of cytokeratins no. 1-3 despite of the cornification which was reliably proved by histology; --the appearance of the proteins having molecular mass values of 42 kDa and less which very probably may be the products of partial keratinolysis as evidenced by immunoblotting with monoclonal and polyclonal antibodies to cytokeratins. Among these proteins a proteolytically modified cytokeratin no. 19 of only 38 kDa was found.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Squamous Cell; Electrophoresis, Polyacrylamide Gel; Humans; Immunoblotting; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Sodium Dodecyl Sulfate

We have used monolayers of bromodeoxyuridine (BrdU)-grown and control human epidermoid carcinoma (A431) cells to investigate the polypeptide changes resulting when cells are rounded by epidermal growth factor (EGF). Whereas no significant change was detected in Triton-soluble components, the urea-solubilized matrix fraction revealed greater levels of a 20 kd species in cells exposed to EGF, compared with the same cells not exposed to the growth factor. The corresponding urea fraction from BrdU-grown cells showed decreased levels of the 20 kd species as a result of exposure to EGF. Further evidence for a differential effect of EGF resulting from growth of the cells with the pyrimidine analog was observed in the matrix fraction soluble in SDS, which revealed a decrease in a 20 kd species resulting from exposure of control cells to EGF, with no comparable effect in BrdU-grown cells. Our results suggest that EGF induces a change in the properties of matrix-associated components of low molecular weight in an effect which appears to be modified by prior growth of cells with BrdU.

Topics: Bromodeoxyuridine; Carcinoma, Squamous Cell; Cell Line; Cytoplasm; Epidermal Growth Factor; Humans; Mercaptoethanol; Octoxynol; Polyethylene Glycols; Proteins; Sodium Dodecyl Sulfate; Solubility; Urea

The purpose of this study is to classify the membrane proteins from some cutaneous tumours (basal cell epitheliomas, squamous epitheliomas, melanomas) and possibly detect the ones which are characteristic of each type of tumour. The microsomal, mitochondrial and nuclear fractions were purified following the usual techniques. The membrane proteins were solubilized with sodium dodecyl sulfate, treated with iodoacetamide and chromatographed in SDS-acrylamide slab-gel.

Topics: Carcinoma, Basal Cell; Carcinoma, Squamous Cell; Chemical Phenomena; Chemistry; Electrophoresis, Polyacrylamide Gel; Humans; Intracellular Membranes; Melanoma; Membrane Proteins; Skin Neoplasms; Sodium Dodecyl Sulfate

The microsomal, mitochondrial, and nuclear fractions of cutaneous tumors have been investigated. Cutaneous tumors were homogenized and microsomal, mitochondrial, and nuclear fractions were purified. The membrane proteins were solubilized with sodium duodecyl sulfate (SDS). The membrane lipids were removed and membrane proteins were solubilized again in a small volume of SDS solution and chromatographed in SDS-acrylamide slab-gel. The plates were stained with Coomassie Brilliant Blue to show protein bands. The preliminary results show that the electrophoretic profiles of microsomal proteins are characteristic of some tumors.

Topics: Carcinoma; Carcinoma, Squamous Cell; Chromatography, Gel; Fibroma; Humans; Intracellular Membranes; Melanoma; Membrane Proteins; Microsomes; Mitochondria; Neoplasm Proteins; Nuclear Envelope; Skin Neoplasms; Sodium Dodecyl Sulfate

Topics: Adenocarcinoma; Antigen-Antibody Complex; Carcinoembryonic Antigen; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Cell Membrane; Cross Reactions; Electrophoresis, Starch Gel; Epitopes; Fluorescent Antibody Technique; Glycoproteins; Humans; Immunodiffusion; Immunoelectrophoresis; Lung Neoplasms; Microsomes; Salicylates; Sodium Dodecyl Sulfate