agar and Lung-Neoplasms

Programmed death ligand-1 (PD-L1) has a known association with the prognosis of human cancers because of its ability to alter tumor immune surveillance via its interaction with PD-1. We questioned whether expression of PD-L1 in tumor cells could directly promote tumor growth and invasiveness in non-small cell lung cancer (NSCLC).. Real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate PD-L1 messenger RNA (mRNA) expression in lung tumors. The prognostic value of PD-L1 mRNA was assessed by Cox regression model. Transcriptional regulation of PD-L1 by human papillomavirus (HPV) 16/18 E6 oncoprotein or by epidermal growth factor receptor (EGFR) mutation in lung cancer cells was examined by Western blot and luciferase reporter assay. The cell growth and invasion were evaluated by colony formation, soft agar growth, and Boyden chamber assay.. The PD-L1 mRNA levels showed a positive association with HPV 16/18 E6 oncoprotein and with EGFR mutation in 223 surgically resected NSCLC patients. The prognostic significance of PD-L1 was more commonly observed in patients with high PD-L1/E6 positive and high PD-L1/EGFR mutant tumors. Mechanistically, upregulation of PD-L1 transcription by E6 or mutant EGFR occurred largely through the ERK-C/EBPβ-TLR4-NF-κB cascade. PD-L1 promotes the efficacy of colony formation, soft agar growth, and cell invasion. PD-L1 upregulates BAG-1 to reduce transforming growth factor (TGF)-β1 expression, and the decrease in SMAD4 because of TGF-β1 occurs through the p53/microRNA (miR)-224 axis. The decreases in TGF-β1 and SMAD4 are responsible for PD-L1-mediated cell invasiveness.. Induction of PD-L1 by E6 oncoprotein or mutant EGFR through the ERK-C/EBPβ-TLR4-NF-κB cascade may promote tumor growth and invasiveness in NSCLC because of decreasing TGF-β1 and SMAD4 expression.

Topics: Agar; B7-H1 Antigen; Carcinoma, Non-Small-Cell Lung; ErbB Receptors; Gene Expression Regulation, Neoplastic; Human papillomavirus 16; Human papillomavirus 18; Humans; Lung Neoplasms; NF-kappa B; RNA, Messenger; Smad4 Protein; Toll-Like Receptor 4; Transforming Growth Factor beta1

Lung tumors express high levels of aromatase enzyme compared to surrounding normal tissue. Inhibition of aromatase has emerged as a recent therapeutic approach for the treatment of breast cancer. However, the role of aromatase inhibition in lung cancer treatment requires further investigation.. The anti-proliferative effects of aromatase inhibitors were evaluated by MTT assay. Cell migration was assessed using a wound healing assay. The mechanism of cell death was determined using the annexin VFITC/ propidium iodide staining flow cytometry method. The soft agar colony formation assay evaluated cells' capability to form colonies.. Exemestane and curcumin significantly inhibited the growth of lung cancer cell lines in a dose- and timedependent manner. The IC. Aromatase inhibition by exemestane or curcumin had significantly inhibited the growth of lung cancer cell lines, synergized with cisplatin, raloxifene, and celecoxib, suppressed lung cancer cell migratory potential, induced apoptosis, and reduced colony formation of lung cancer cells.

Topics: Agar; Annexins; Apoptosis; Aromatase; Aromatase Inhibitors; Celecoxib; Cell Line, Tumor; Cell Proliferation; Cisplatin; Curcumin; Humans; Lung Neoplasms; Propidium; Raloxifene Hydrochloride

Kirsten rat sarcoma viral oncogene homolog (KRAS) aberrations frequently occur in patients with lung cancer. Oncogenic KRAS is characterized by excessive reactive oxygen species (ROS) accumulation, thus, ROS detoxification may contribute to KRAS‑driven lung tumorigenesis. In the present study, the influence of glutathione peroxidase 2 (GPX2) on malignant progression and cisplatin resistance of KRAS‑driven lung cancer was explored. The RNA sequencing data from TCGA lung cancer samples and GEO database were downloaded and analyzed. The effects of GPX2 on KRAS‑driven lung tumorigenesis were evaluated by western blotting, cell viability assay, soft agar assay, Transwell assay, tumor xenograft model, flow cytometry, BrdU incorporation assay, transcriptome RNA sequencing, luciferase reporter assay and RNA immunoprecipitation. In the present study, GPX2 was upregulated in patients with non‑small cell lung carcinoma (NSCLC), and positively correlated with poor overall survival. Ectopic GPX2 expression facilitated malignant progression of KRAS

Topics: Agar; Bromodeoxyuridine; Carcinogenesis; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Cell Proliferation; Cisplatin; Drug Resistance, Neoplasm; Glutathione Peroxidase; Humans; Lung; Lung Neoplasms; Matrix Metalloproteinase 1; MicroRNAs; Proto-Oncogene Proteins p21(ras); Reactive Oxygen Species

Quick and reliable testing of EGFR and KRAS is needed in non-small cell lung cancer (NSCLC) to ensure optimal decision-making for targeted therapy. The Idylla™ platform was designed for Formalin-Fixed Paraffin-Embedded (FFPE) tissue sections but recently several studies were published that evaluated its potential for cytological specimens. This study aimed to validate the Idylla™ platform for the detection of EGFR/KRAS mutations in cytological NSCLC samples prepared as cytoblocks using AGAR and paraffin embedding.. The KRAS Idylla™ test were performed on 11 specimens with a known KRAS mutation. The EGFR Idylla™ test was performed on 18 specimens with a known primary EGFR mutation and 7 specimens with a primary EGFR-EGFR T790M resistance mutation combination.. Concordant KRAS and primary EGFR mutations were detected for both KRAS and primary EGFR mutations. Samples with a total CQ value of < 26 could be considered negative. Samples with a total CQ value of > 26 could not be assessed (probability of false-negative). In specimens with a primary EGFR-EGFR T790M resistance mutation combination, 5/7 cases were not concordant.. Our results confirm the conclusion of recent reports that the Idylla™EGFR assay is not suitable in a resistance to EGFR TKI setting, also not in our cytological NSCLC samples prepared as cytoblocks using AGAR and paraffin embedding. KRAS and primary EGFR mutations were detected using the Idylla™ assays in virtually all cytological NSCLC samples. This analysis was rapid and time-saving compared to other mutation detection assays and may be useful if the amount of material is insufficient to perform a full set of molecular tests.

Topics: Adenocarcinoma; Agar; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Fixatives; Formaldehyde; Genes, erbB-1; Genetic Testing; Humans; Lung Neoplasms; Mutation; Paraffin Embedding; Proto-Oncogene Proteins p21(ras); Real-Time Polymerase Chain Reaction; Tissue Fixation

We report a case of canine adenocarcinoma with multi-organ metastasis in which colonies of adenocarcinoma cells grew upon aerobic bacterial culture of pleural effusion. Stained agar colonies were highly similar to rare suspicious cells seen on cytologic examination of the pleural effusion, as well as rare cells seen on cytologic examination of pancreatic and gastric wall fine-needle aspirates. Cells from colonies growing on agar media were mildly immunoreactive for cytokeratin. Histologic examination of tissues obtained at autopsy revealed pancreatic adenocarcinoma with vascular invasion and nodal, gastric, pulmonary, and pleural metastasis.

Topics: Adenocarcinoma; Agar; Animals; Bacteriological Techniques; Biopsy, Fine-Needle; Culture Media; Dog Diseases; Dogs; Female; Lung; Lung Neoplasms; Lymphatic Metastasis; Pancreatic Neoplasms; Pleural Effusion, Malignant; Pleural Neoplasms; Stomach Neoplasms

The spread of lung cancer cells to distant sites represents a common event associated with poor prognosis. A fraction of tumor cells named cancer stem cells (CSCs) have the ability to overcome therapeutic stress and remain quiescent. However, whether these CSCs have also the capacity to initiate and sustain metastasis remains unclear. Here, we used tumor sphere cultures (TSC) isolated from mouse and human lung cancer models to enrich for CSCs, and assessed their metastatic potential as compared to non-CSCs. As expected, TSC overexpressed a variety of stem cell markers and displayed chemoresistance. The CSC phenotype of TSC was confirmed by their higher growth ability in soft agar and tumorigenic potential in vivo, despite their reduced in vitro cell growth kinetics. Surprisingly, the appearance of spontaneous lung metastases was strongly delayed in mice injected with TSC as compared to non-TSC cells. Similarly, this finding was confirmed in several other models of metastasis, an effect associated with a retarded colonization activity. Interestingly, such delay correlated with a quiescent phenotype whose underlined mechanisms included an increase in p27 protein and lower phospho-ERK1/2 levels. Thus, these data suggest that cells enriched for CSC properties display an impaired metastatic activity, a finding with potential clinical implications.

Topics: Agar; Animals; Antineoplastic Agents; Cell Proliferation; Cyclin-Dependent Kinase Inhibitor p27; Drug Resistance, Neoplasm; Extracellular Signal-Regulated MAP Kinases; Female; Humans; Lung Neoplasms; Mice; Mice, Nude; Mice, Transgenic; Neoplasm Metastasis; Neoplastic Stem Cells; Osteolysis; p38 Mitogen-Activated Protein Kinases; Phenotype; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Spheroids, Cellular

The matrix glycoprotein, fibronectin, stimulates the proliferation of non-small cell lung carcinoma in vitro through α5β1 integrin receptor-mediated signals. However, the true role of fibronectin and its receptor in lung carcinogenesis in vivo remains unclear. To test this, we generated mouse Lewis lung carcinoma cells stably transfected with short hairpin RNA shRNA targeting the α5 integrin subunit. These cells were characterized and tested in proliferation, cell adhesion, migration, and soft agar colony formation assays in vitro. In addition, their growth and metastatic potential was tested in vivo in a murine model of lung cancer. We found that transfected Lewis lung carcinoma cells showed decreased expression of the α5 gene, which was associated with decreased adhesion to fibronectin and reduced cell migration, proliferation, and colony formation when compared with control cells and cells stably transfected with α2 integrin subunit in vitro. C57BL/6 mice injected with α5-silenced cells showed lower burden of implanted tumors, and a dramatic decrease in lung metastases resulting in higher survival as compared with mice injected with wild-type or α2 integrin-silenced cells. These observations reveal that recognition of host- and/or tumor-derived fibronectin via α5β1 is important for tumor growth both in vitro and in vivo, and unveil α5β1 as a potential target for the development of anti-lung cancer therapies.

Topics: Agar; Animals; Carcinoma, Lewis Lung; Cell Adhesion; Cell Movement; Cell Proliferation; Disease Progression; Fibronectins; Gene Silencing; Injections, Intravenous; Injections, Subcutaneous; Integrin alpha5beta1; Lung Neoplasms; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neoplasm Transplantation; RNA, Small Interfering; Survival Analysis; Tumor Stem Cell Assay

The purpose of this study was to validate the accuracy and reliability of volume measurements obtained using three-dimensional (3D) thoracoscopic ultrasound (US) imaging. Artificial "tumours" were created by injecting a liquid agar mixture into spherical moulds of known volume. Once solidified, the "tumours" were implanted into the lung tissue in both a porcine lung sample ex vivo and a surgical porcine model in vivo. 3D US images were created by mechanically rotating the thoracoscopic ultrasound probe about its long axis while the transducer was maintained in close contact with the tissue. Volume measurements were made by one observer using the ultrasound images and a manual-radial segmentation technique and these were compared with the known volumes of the agar. In vitro measurements had average accuracy and precision of 4.76% and 1.77%, respectively; in vivo measurements had average accuracy and precision of 8.18% and 1.75%, respectively. The 3D thoracoscopic ultrasound can be used to accurately and reproducibly measure "tumour" volumes both in vivo and ex vivo.

Topics: Agar; Algorithms; Animals; Automation; Humans; Image Processing, Computer-Assisted; Imaging, Three-Dimensional; Lung; Lung Neoplasms; Neoplasm Transplantation; Phantoms, Imaging; Radiography; Reproducibility of Results; Swine; Ultrasonography

Manganese superoxide dismutase (MnSOD) activity is generally lower in cancer cells when compared with their normal cell counterparts. Many studies have shown that replacing the diminished MnSOD activity leads to inhibition of the malignant phenotype. We sought to overexpress MnSOD in a chemically transformed, malignant rat cell line with low endogenous MnSOD activity to determine the effect on the malignant phenotype. After MnSOD cDNA transfection, clonal populations were characterized at the molecular level for protein, RNA, and DNA, as well as for in vitro and in vivo growth and in vivo lung metastasis. MnSOD transfectants, which both under- and overexpressed MnSOD protein, were identified. These transfectants demonstrated variations in glutathione peroxidase and catalase activity levels, indicating differences in peroxide-generating versus peroxide-metabolizing enzymes (antioxidant imbalance); these differences were suggestive of alterations in their abilities to metabolize peroxide when compared with the parental cell line. In addition, these transfectants demonstrated reductions in both in vitro and in vivo growth, as well as a reduction in metastatic potential, which correlated with antioxidant imbalance. These results suggest that the tumor suppressive effect of MnSOD overexpression is in part mediated by an antioxidant imbalance resulting in the reduced capacity to metabolize increased levels of intracellular peroxides.

Topics: Agar; Animals; Antioxidants; Blotting, Southern; Blotting, Western; Catalase; Cell Line, Transformed; Cell Line, Tumor; Chloramphenicol O-Acetyltransferase; DNA; DNA, Complementary; Glutathione Peroxidase; Lung Neoplasms; Mice; Mice, Nude; Neoplasm Metastasis; Peroxides; Phenotype; Plasmids; Rats; Reverse Transcriptase Polymerase Chain Reaction; RNA; Superoxide Dismutase; Time Factors; Transfection

Small pulmonary lesions with ground-glass opacity (GGO) are increasingly detected by CT; however, intraoperative localization of such lesions is difficult because these lesions are often invisible and nonpalpable.. To localize and resect nonpalpable and invisible small pulmonary lesions, a new marking technique that we call "agar marking" was developed.. Powdered agar was dissolved in distilled water at a concentration of 5% and kept at > 50 degrees C to maintain its liquid form. Agar was injected through an 18-gauge needle and placed near the target lesion with CT. After animal experiments, agar marking was applied to the nine patients who had lesions < 20 mm in diameter and lesions with GGO. The mean diameter of these lesions was 11 mm, with a mean depth of 19 mm from the pleural surface on CT.. Agar could be detected as a hard nodule by manual palpation, and the lesion was resected during thoracotomy in all cases. There were no complications associated with the agar injection, aside from one case of slight pneumothorax.. Agar marking may represent a feasible alternative technique for localizing nonpalpable occult lesions located away from the pleural surface.

Topics: Agar; Aged; Animals; Dogs; Female; Humans; Intraoperative Care; Lung; Lung Neoplasms; Male; Middle Aged; Palpation; Solitary Pulmonary Nodule; Tomography, X-Ray Computed

Small cell lung cancer (SCLC) is a highly invasive and metastatic tumor, and the decreased expression of alpha3beta1 integrin may contribute to its virulence. Alpha3beta1 is a critical integrin for pulmonary development and epithelial integrity, and its reduced expression has been linked to the increased malignancy and invasion of other cancers. The amplification of the c-myc oncogene is seen frequently in relapsed SCLC tumors and is associated with a worsened prognosis. In the present study using a model of SCLC tumor progression, overexpression of c-myc in a classic SCLC cell line, NCI H209, enhanced in vitro features of tumorigenesis, altered the relationships between cell and environment, and markedly down-regulated the expression of the alpha3 integrin subunit at both the transcript and protein levels. This inverse relationship between the expression of the alpha3 integrin subunit and c-myc is mimicked by other c-myc-overexpressing SCLC cell lines. Restoring alpha3 expression in the myc-transfected 209 cells reversed the effects of c-myc: alpha3 transfection increased cell:cell adhesion and reduced soft agar cloning without affecting the in vitro doubling time. The diminished soft agar cloning produced by alpha3 transfection was reversed by an antibody that specifically engages alpha3beta1 integrins, P1B5. These results suggest first, that alpha3beta1 integrin mediates homotypic adhesion of SCLC cells, and second, that unengaged alpha3beta1 integrin suppresses the growth of disaggregated SCLC cells. Thus, the down-regulation of the alpha3 integrin subunit may contribute to the enhanced tumorigenicity of c-myc-overexpressing SCLCs by allowing the growth of tumor cells that have reduced contact with ligand-expressing substratum or cells, a condition that occurs during the growth of the primary tumor, tumor invasion, and metastasis.

Topics: Agar; Antibodies; Antigens, CD; Carcinoma, Small Cell; Cell Adhesion; Cell Aggregation; Clone Cells; Disease Progression; Gene Expression; Genes, myc; Humans; Integrin alpha3; Integrin alpha3beta1; Integrins; Lung Neoplasms; Proto-Oncogene Proteins c-myc; Transfection; Tumor Cells, Cultured

Lung cancers have been distinguished into small-cell lung cancer (SCLC) and non-small cell-lung cancer (NSCLC) types on the basis of their clinical behaviors and their responses to treatment. Moreover, growth of most SCLC cell lines in liquid culture medium is nonadherent, while that of most NSCLC cell lines is adherent. In this study, we examined the effect of matrigel (reconstituted basement membrane components), which is known to have growth-stimulatory activity on various human tumor cell lines in immunodeficient mice, on soft-agar colony formation of a panel of SCLC and NSCLC cell lines to clarify its mechanism of growth stimulation of cancer cells. Matrigel enhanced colony formation of all 9 NSCLC cell lines and 4 of 9 SCLC cell lines. There was a statistically significant difference (P < 0.01) between colony formations with and without matrigel of NSCLC cell lines, but not for SCLC cell lines. In liquid culture medium, all 9 NSCLC lines and 3 of 9 SCLC lines adhered to plastic dishes, whereas the other SCLC lines did not. Matrigel enhanced colony formation of all 3 adherent-type SCLC lines and 1 of 6 nonadherent-type NSCLC lines. Matrigel enhanced colony formation of both of 2 adherent-type non-lung cancer cell lines and 1 of 2 nonadherent-type leukemia cell lines. Neither transforming growth factor beta, collagen type IV, fibronectin, nor laminin, which are components of matrigel, enhanced colony formation of an NSCLC cell line in soft agar. The increase in the colony number of the NSCLC cell line by matrigel was abrogated by the protein kinase inhibitors staurosporine and UCN-01.

Topics: Adult; Agar; Aged; Aged, 80 and over; Biocompatible Materials; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Collagen; Drug Combinations; Female; Humans; Laminin; Lung Neoplasms; Male; Middle Aged; Proteoglycans; Tumor Cells, Cultured

Insulin-like growth factors (IGFs) are often essential for the maintenance of the malignant phenotype, and in lung cancer the IGF-I receptor (IGF-Ir) is often expressed at high levels. Stable transfection of antisense plasmids expressing the first 300 bp of the IGF-Ir reduces the tumorigenicity of a variety of tumor cell lines and has been reported to induce systemic antitumor effects on established, non-gene-modified tumors in animal model systems. We have constructed an adenovirus expressing an antisense IGF-Ir (Ad-IGF-Ir/as) in an attempt to develop these observations into a clinical therapeutic approach. A single transduction by Ad-IGF-Ir/as (at a multiplicity of infection of 10:1) decreased the IGF-Ir number by about 50% in human lung cancer cell lines NCI H460 and SCC5, as measured by an 125I-labeled IGF-I competitive binding assay. After the transduction of these human lung cancer cell lines by Ad-IGF-Ir/as, the soft agar clonogenicity was reduced by 84%. The i.p. treatment of nude mice bearing established i.p. NCI H460 cells resulted in prolonged survival compared to that of nude mice treated with a reporter virus. These results suggest that Ad-IGF-Ir/as has a therapeutic effect on established human lung cancer xenografts and may represent an effective and practical cancer gene therapy strategy.

Topics: Adenoviridae; Agar; Animals; Base Sequence; Binding, Competitive; Carcinoma, Small Cell; Cell Division; Clone Cells; Female; Humans; Insulin-Like Growth Factor I; Iodine Radioisotopes; Lung Neoplasms; Mice; Mice, Nude; Molecular Sequence Data; Neoplasm Transplantation; Oligonucleotides, Antisense; Receptor, IGF Type 1; Transduction, Genetic; Transfection; Transplantation, Heterologous; Tumor Cells, Cultured

3-Hydroxy-3-methylglutaryl CoA reductase (HMG-CoA reductase) plays a rate-limiting role in isoprenoid biosynthesis and is associated with cell proliferation and transformation. Although an elevated level of HMG-CoA reductase activity is consistently detected in cancer cell lines and tumors, the question remains whether HMG-CoA reductase activity may have a causative role in cell transformation. We have stably transfected the A549 human adenocarcinoma cells with both bicistronic and retroviral expression vectors, including the whole cDNA of human HMG-CoA reductase. Stably transfected cells showed strong morphological changes and disorganization in the filamentous actin architecture, became contact inhibited, and had a lower doubling time. Moreover, they exhibited anchorage-independent growth reduction and lost their capability to induce tumors in nude mice. Surprisingly, no quantitative modification of enzyme activity was observed following transfection, although expression of HMG-CoA reductase cDNA was shown by Northern blot analysis. When endogenous and transfected reductase activity was bypassed by the addition of mevalonate and compactin, a competitive inhibitor, the filamentous actin distribution in HMG-CoA reductase-transfected cells became very similar to that of control cells, demonstrating the role of exogenous HMG-CoA reductase activity in this process. All of our data together strongly suggest that phenotype reversion is dependent on exogenous HMG-CoA reductase expression and that enzymatic activity is implied in this mechanism. HMG-CoA reductase cDNA expression, by expression of a particular form of reductase, might be a negative regulator of cell growth and thus reverse the phenotype of tumor cells.

Topics: Acyl Coenzyme A; Adenocarcinoma; Agar; Animals; Cell Division; Cell Line, Transformed; Cloning, Molecular; Cytoskeleton; DNA, Complementary; Humans; Kinetics; Lung Neoplasms; Mice; Mice, Nude; Oxidoreductases; Phenotype; RNA, Messenger; Transformation, Genetic; Tumor Cells, Cultured

Soft agar clonogenic assays are considered to be a standard method for measuring tumour cell radiosensitivity and it has been widely reported that fibroblast contamination does not occur. We report here that human fibroblasts can proliferate to form colonies in a modified form of the Courtenay-Mills soft agar clonogenic assay. It was observed that early passage skin fibroblasts could form colonies in soft agar, although the plating efficiencies were reduced compared with growth on plastic. It was demonstrated that normal lung could proliferate in agar with similar plating efficiencies to fresh tumours and that fibroblastic cells were present in these cultures. Characterisation of primary lung tumour cultures also showed that fibroblastic cells were present in these cultures. Characterisation of primary lung tumour cultures also showed that fibroblastic cells were present which lacked epithelial features and which resembled closely the cells found in cultures of normal lung. This is an important finding for workers using soft agar assays to culture human tumour cells and is of interest in understanding the processes of normal growth control of human fibroblasts.

Topics: Agar; Cell Division; Cells, Cultured; Culture Media; Epithelial Cells; Epithelium; Fibroblasts; Humans; Keratins; Lung; Lung Neoplasms; Microscopy, Electron; Radiation Tolerance; Skin; Tumor Cells, Cultured; Tumor Stem Cell Assay

The ultrastructure of cells from seven human lung cancers and from the colonies formed by these cells in soft agar was investigated. Tumor cells developed to display the morphofunctional potentials of the initial tumors. Cultured cells of squamous-cell carcinomas contained numerous tonofilaments, those of adenocarcinomas developed microvilli on their apical surfaces and intracellular lumens. On the other hand, cells of squamous-cell carcinomas showed features specific of adenoma epithelium, i.e. well developed microvilli and intracellular lumens. Besides, cells of adenocarcinoma often contained large quantities of tonofilaments considered to be characteristic of epidermoid epithelium. The results obtained suggest a possibility of metaplastic transformation of the lung epithelium.

Topics: Adenocarcinoma; Adenocarcinoma, Bronchiolo-Alveolar; Agar; Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Culture Media; Humans; Lung Neoplasms; Methotrexate; Microscopy, Electron; Morphogenesis; Prospidium; Tumor Cells, Cultured; Vincristine

We studied the effects human recombinant granulocyte-macrophage colony-stimulating factor and human recombinant interleukin-3 on the colony formation of three human solid tumor cell lines. Using a modified double-layer soft agar clonogenic assay rhGM-CSF enhanced colony formation of all cell lines tested in a dose dependent manner (up to twofold for the breast cancer cell line BT-20, up to 163% of the control for the hypernephroma cell line C 94 and up to 147% for the non-small cell lung cancer cell line CCL 185 at a concentration of 100 ng/ml). RhIL-3 stimulated colony formation of the cell lines C 94 and BT-20, whereas on the cell line CCL 185 rhIL-3 had no effect even at the highest dose level tested (100 ng/ml). Combinations of growth factors showed subadditive stimulation on two cell lines tested (BT-20, C 94). These data indicate that haematopoietic growth factors exert a growth promoting activity on certain solid tumor cells in vitro at physiological concentrations. Therefore our results suggest that the application of these factors in immuno- and myelosuppressed cancer patients after high dose chemotherapy should be seen in light of a possible co-stimulation of the malignant cells.

Topics: Agar; Breast Neoplasms; Carcinoma, Non-Small-Cell Lung; Carcinoma, Renal Cell; Cell Division; Colony-Stimulating Factors; Granulocyte-Macrophage Colony-Stimulating Factor; Growth Substances; Humans; Interleukin-3; Kidney Neoplasms; Lung Neoplasms; Neoplastic Stem Cells; Recombinant Proteins; Statistics as Topic; Tumor Cells, Cultured; Tumor Stem Cell Assay

The mouse fibroblast-like transformed cell line C-243 adapted to growth in suspension was used as a source of virus-induced interferons (MulFN-alpha, beta), and spontaneously produced active growth factors. These factors were purified from C-243 cells grown as tumors in BALB/c mice, and had properties identical to those of TGF-alpha or TGF-beta isolated by others from different tissues. Exogenous TGF-alpha, beta stimulated colony formation by C-243 cells in soft agar, whereas MulFN-alpha, beta inhibited it. Clonal growth of human lung adenocarcinoma A549 cells in soft agar was inhibited as well by human interferons (types alpha, beta, or gamma) as by TGF-beta. Inhibition was dose-related. Pure EGF, which is an analogue of TGF-alpha, diminished the antiproliferative activity of interferons alpha, beta, and gamma in A549 cells. On the other hand, the anti-mitogenic action of IFN-beta and TGF-beta was clearly synergistic. In mice bearing C-243 cell tumors, TGF-alpha, beta stimulated growth, whereas MulFN-alpha, beta inhibited it. Stimulation of tumor growth was also observed after administration of anti-IFN serum that could neutralize endogenous IFN-alpha, beta. The simultaneous administration of MulFN-alpha, beta and TGF-alpha, beta diminished anti-tumor effects of IFN in mice. Our results suggest that both TGFs and IFNs are autocrine, positive or negative growth factors modulating the rate of proliferation and the neoplastic behavior of the cells. The final effects depend on the target-cell sensitivity and on the relative concentration of the various hormone-like factors. Cancer cells overstimulated by TGF-alpha, beta or by EGF may not respond to IFNs.

Topics: Adenocarcinoma; Agar; Animals; Cell Division; Clone Cells; Humans; Interferons; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Peptides; Transforming Growth Factors

The human tumor clonogenic assay (HTCA) has potential value for studies of both the chemosensitivity and biology of human tumors. However, many technical problems including low plating efficiencies and the preparation of sufficient numbers of viable cells remain. In this study, an improved method for disaggregation of solid tumors increased the yield of single cells. Consequently, more than 10 anticancer drugs could be tested in 94 of 168 specimens (56%). Removal of peripheral blood lymphocytes from cell suspensions derived from effusions also improved colony formation. Adequate growth for sensitivity testing (greater than 30 colonies/plate) was obtained in 122 cases (73%), inadequate growth for drug evaluation (5-29 colonies/plate) in 29 cases (17%), and no colony formation (less than 5 colonies/plate) in 17 cases (10%) of the 168 viable samples. The cloning efficiencies of cells derived from primary tumors (median 0.015%) were higher than those of cells derived from metastatic tumors (0.012%), and they varied with the location of the metastatic site. Cloning efficiencies varied markedly from specimen to specimen, and were unaffected by tumor histology, grade of differentiation, patient age, stage of disease, or prior chemotherapy. The HTCA is promising as a potential tool for studying the biology of tumors.

Topics: Agar; Carcinoma; Cell Aggregation; Colony-Forming Units Assay; Humans; Lung Neoplasms; Neoplasm Staging; Prognosis; Tumor Stem Cell Assay

Spontaneous primary mammary tumors of C3H-Avy mice differ in metastatic colonization potential, some producing many lung deposits (high-colonization potential) and others producing few or none (low-colonization potential) after iv inoculation of cells. The degree of metastasis from undisturbed neoplasms also varies from tumor to tumor. This study examined whether these differences between tumors could be accounted for by differences in clonogenic or stem cell content. Tests for clonogenic cells were: growth in 0.3% agarose and limiting dilution assays. Mammary tumor cells of known colonization potential were inoculated iv at serially reduced doses, and the relationship between number of cells injected and number of lung deposits formed was determined. Parallel in vitro dose-response assays in 0.3% agarose were performed with the use of cells from the same primary tumors. Colony-forming efficiency in 0.3% agarose cultures varied between individual primary mammary tumors and was positively associated with experimental metastatic potential, suggesting that the stem or clonogenic cell content of primary tumors is one of the important determinants of the metastatic phenotype.

Topics: Agar; Animals; Colony-Forming Units Assay; Culture Media; Female; Lung Neoplasms; Mammary Neoplasms, Experimental; Mice; Mice, Inbred C3H; Neoplasm Metastasis; Sepharose; Tumor Stem Cell Assay

Topics: Agar; Animals; Cell Division; Cell Transformation, Neoplastic; Cells, Cultured; Clone Cells; Lung Neoplasms; Mammary Neoplasms, Experimental; Mice; Neoplasm Metastasis; Neoplastic Stem Cells; Rats; Tumor Stem Cell Assay

A continuous human cell line was established from a patient with large cell anaplastic lung carcinoma. This cell line, designated QU-DB, has been in culture for over 36 months and grows as an adherent monolayer with a doubling time of 10-12 hours. Its morphology, ultrastructure, karyotype, ability to grow in soft agar and heterotransplantability, indicate it is a large-cell lung tumor cell line of human origin. Three cell lines were established from metastatic tumors in nude mice receiving subcutaneous injections of QU-DB cells. The morphology and growth characteristics exhibited by these cell lines were similar to the primary cell line. Karyotypic analysis of cell lines derived from the primary tumor and a metastasis to the diaphragm were similar, but cells from a liver metastasis culture showed additional karyotypic changes. This large cell lung tumor cell line may prove useful as a model system for studies of human tumor progression and metastasis.

Topics: Agar; Aged; Animals; Carcinoma; Cell Division; Cell Line; Clone Cells; Flow Cytometry; Humans; Karyotyping; Liver Neoplasms; Lung Neoplasms; Male; Mice; Mice, Nude; Microscopy, Electron; Neoplasm Metastasis; Smoking

In C57Bl/6 mice with different stages of Lewis lung carcinoma growth used as recipients of diffusion chambers, after subcutaneous transplantation of tumour cells there was an increased output of colonies in the bone marrow and spleen cell cultures (CFU-DC) as compared to the control, The cocultivation of normal (bone marrow and spleen) and tumour cells has no influence on the effectiveness of their cloning.

Topics: Abdomen; Agar; Animals; Bone Marrow; Bone Marrow Cells; Carcinoma; Cell Count; Cells, Cultured; Diffusion; Hematopoietic Stem Cells; Lung Neoplasms; Mice; Mice, Inbred C57BL; Spleen; Time Factors

A retrovirus isolated from experimentally induced sheep lung carcinoma (SPCTV) was propagated in chronically infected Himalayan tahr ovarian cells and in normal sheep lung cells. Follow-up of infection of the cells with SPCTV showed the appearance of syncytium, plaque formation, partial recovery and the establishment of a chronic infection. Virus-associated reverse transcriptase activity in the medium fluctuated but remained at a constantly high level at the stage of chronic infection. Stages of type-C virus morphogenesis were demonstrated by electron microscopy. The viral genome was detected in both the nucleus and cytoplasm by in situ hybridization. Chronically infected cells formed colonies when plated in soft agar. Following subcutaneous inoculation of chronically infected cells (of fibroblast origin) into nude mice, lymphoid tumors developed at the site of inoculation and in vital organs.

Topics: Agar; Animals; Cell Line; Cell Transformation, Viral; Cytopathogenic Effect, Viral; Female; Lung; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Ovary; Retroviridae; RNA-Directed DNA Polymerase; RNA, Viral; Sheep; Sheep Diseases; Tumor Virus Infections; Virion; Virus Replication

Three distinct dissemination-related phenotypes have been distinguished among cell subpopulations of the mouse B16 melanoma: tumorigenicity, spontaneous metastasis from subcutaneous tumors, and organ colonization following intravenous injection of cells. From a progenitor clone (G3) of tumorigenic but nonmetastatic and noncolonizing (null) cells that underwent phenotypic diversification in vitro and in vivo, 4 subclones were obtained: G3.5 (culture-generated metastatic), G3.12 (tumor-generated metastatic), G3.15 (culture-generated null), and G3.26 (tumor-generated colonizing). The growth potentials of the parent clone and derived subclones were investigated comparatively in in vivo assays (tumorigenicity, tumor growth rate, and lung colonization potential), monolayer culture assays (generation time, saturation density, clonogenicity, and rate of detachment by trypsin), and in soft agar. In overall growth potential, G3.26 greater than G3.12 greater than G3, G3.5 greater than G3.15. These results indicate that metastatic populations of the B16 melanoma are not the most rapidly and effectively growing cells obtainable from that tumor.

Topics: Agar; Animals; Cell Division; Cell Movement; Cells, Cultured; Clone Cells; Female; Lung Neoplasms; Melanoma; Mice; Mice, Inbred C57BL; Microscopy, Phase-Contrast; Models, Biological; Neoplasm Metastasis; Neoplasm Transplantation; Neoplastic Stem Cells; Stem Cells

The photosensitizing/cytotoxic effects of four porphyrin compounds derived from hematoporphyrin were compared using two human tumor cell lines with a soft-agar colony-forming assay. Reproducible dose- and time-dependent increases in reproductive cell death were observed for each porphyrin tested. Two fractions derived from hematoporphyrin derivative (HPD) were found to be better photosensitizers than HPD itself, indicating the potential value of this in vitro assay for detecting the most active component from a heterogeneous mixture.

Topics: Agar; Cell Line; Cell Survival; Colony-Forming Units Assay; Hematoporphyrin Derivative; Hematoporphyrins; Humans; Light; Lung Neoplasms; Melanoma; Radiation-Sensitizing Agents

Topics: Agar; Biopsy; Breast Neoplasms; Cell Survival; Clone Cells; Culture Techniques; Female; Humans; Lung Neoplasms; Male; Melanoma; Methylcellulose; Microscopy, Electron; Neoplasms; Pleural Effusion

A method was developed for cloning the Lewis lung carcinoma cells (LLC) in vivo in diffusion chambers. This method is analogous to the system introduced by Gordon for cloning hemopoietic precursor cells. No significant differences were observed in the efficiency of the cloning of the tumour cells within the range of 1 to 4 weeks after transplantation (7.7 +/- 0.7% for 240 cultures).

Topics: Agar; Animals; Clone Cells; Culture Techniques; Diffusion; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Neoplasm Transplantation; Neoplastic Stem Cells; Stem Cells

Sixteen histologically documented testicular cancer specimens obtained at diagnostic procedures following induction chemotherapy with cis-platinum containing regimens were cloned in soft agar. Seven (44%) of the specimens cultured formed colonies with a mean cloning efficiency of .021%. Colony formation was observed with all the common histologic subtypes of testicular cancer (seminoma, embryonal carcinoma, choriocarcinoma and mixed tumors). In vitro drug sensitivity tests were performed using cis-platinum, vinblastine and VP-16. Three of four specimens demonstrated a decrease in colony formation to less than 50% of controls after a 1 h exposure to VP-16 at 300 micrograms/ml. Two of these patients had a response to treatment with a VP-16 based salvage regimen. Immunoperoxidase staining of the colonies for alpha feto protein and human chorionic gonadotropin were correlated with the serum levels of these tumor markers determined at the time the specimen was obtained. In three instances the same markers were elevated in the serum as detected within cells which formed the colonies; however, in two other cases the marker(s) that was elevated in the serum was not expressed in the colonies. In one case a biopsy of a residual retroperitoneal mass following chemotherapy histologically was a teratoma, but it formed colonies in the assay which stained positive for alpha feto protein. This patient subsequently developed an elevated serum alpha feto protein. These studies have demonstrated that (a) testicular cancer can be cloned directly in soft agar; (b) a heterogeneous tumor cell population exists in metastatic testicular cancer specimens; and (c) a dose response exists for VP-16 in relapsed testicular cancer which suggests that increasing the dose of VP-16 may be clinically beneficial.

Topics: Agar; alpha-Fetoproteins; Chorionic Gonadotropin; Cisplatin; Clone Cells; Dose-Response Relationship, Drug; Etoposide; Humans; Lung Neoplasms; Lymphatic Metastasis; Male; Retroperitoneal Neoplasms; Testicular Neoplasms; Vinblastine

The correlation of the colony growth of cells disaggregated from human melanoma, sarcoma, lung, and ovarian carcinomas were studied in four different semisolid tissue culture assays: (a) the soft agar assay of Pluznik and Sachs; (b) the soft agar assay of Hamburger and Salmon; (c) the soft agar-methyl cellulose assay of Buick et al.; and (d) the methyl cellulose assay of Ogawa et al. There was no colony growth of tumor cells achieved in 15 of 15 cases assayed in Ogawa's methyl cellulose assay. The plating efficiency of the above mentioned tumors was similar in the assays of Pluznik and Sachs, Hamburger and Salmon, and Buick et al. However, the tumor take rate differed among these three systems. The assay of Buick et al. appears potentially useful for analysis of the biology of human tumors.

Topics: Adenocarcinoma; Agar; Cell Division; Cells, Cultured; Cytological Techniques; Female; Humans; Lung Neoplasms; Melanoma; Methylcellulose; Ovarian Neoplasms; Sarcoma

The factors controlling the growth of human tumor cells in soft agar are poorly understood. However, it has been demonstrated that serum provides factors which promote anchorage-independent growth. We tested 58 tumor specimens, which were obtained from patients with adenocarcinoma of the lung, colon, ovary or squamous cell carcinoma, for their ability to form colonies in soft agar in serum-free or serum-supplemented media. The cells were unable to replicate, and none of the hormones or growth factors tested: insulin (I), transferrin (T), selenium (S), estradiol (E), hydrocortisone (H) or epidermal growth factor (EGF) could substitute for serum. Examination of the serum dose-response curves indicated that growth factors reduced the serum concentrations needed to support anchorage-independent growth. The addition of the supplements and the lowering of serum concentrations increased cloning efficiencies (C.E.) in 38/51 trials, when cells were able to grow initially. The addition of ITS increased C.E. in 18/21 cases, HITES in 15/17 cases and EGF in 12/18 cases as compared to controls. ITS and HITES increased the number of colonies only when serum was the limiting factor. EGF, however, increased the number of colonies even when serum was not the limiting factor. The ability of the supplements to enhance growth could not be correlated to tumor type or initial cloning efficiencies. However, in only 1/25 cases were cells that were unable to form colonies under standard conditions induced to form colonies in the presence of the growth factors. Normal and tumor-derived human fibroblasts did not form colonies in soft agar in the presence of these growth factors. The results suggest that human tumor cells may require the presence of serum-derived factors for growth in soft agar.

Topics: Adenocarcinoma; Agar; Blood; Carcinoma, Squamous Cell; Cell Division; Clone Cells; Colonic Neoplasms; Epidermal Growth Factor; Estradiol; Female; Humans; Hydrocortisone; Insulin; Lung Neoplasms; Ovarian Neoplasms; Selenium; Transferrin

We have compared the sensitivities of two methodologies for determining bone marrow involvement by small cell lung cancer. These methodologies included histological examination of marrow aspirations and biopsies versus growth of tumor colonies in soft agar. There were four instances in which histological study of the marrow aspirate (and biopsy) revealed metastatic small cell lung cancer. All four of the specimens formed colonies in soft agar. Thirty-four of 37 histologically negative aspirations and biopsies) showed no growth in the soft agar system. However, three histologically negative specimens formed colonies in soft agar. The cells growing in these colonies were documented to be small cell lung cancer by histology and growth in nude mice. We conclude that small cell lung cancer metastatic to bone marrow will form colonies in soft agar. Additional study is needed to determine if the soft agar system is indeed more sensitive than routine histology in detecting small cell lung cancer metastatic to bone marrow.

Topics: Agar; Bone Marrow; Carcinoma, Small Cell; Cells, Cultured; Clone Cells; Culture Media; Humans; Lung Neoplasms

Seventeen well-characterized human lung cancer cell lines were examined for the presence of specific membrane receptors for epidermal growth factor (EGF) and nerve growth factor (NGF) as well as for the production of diffusible factors capable of stimulating soft agar growth. These cell lines represented all four major histological types of human lung cancer including small cell carcinoma of the lung (SCCL) and the three types of non-SCCL (epidermoid, large cell, and adenocarcinoma). The SCCL lines included three lines referred to as "converters" because they had lost SCCL morphological and biochemical properties during prolonged passage in vitro. Specific receptors for EGF and NGF were detected by measuring the binding of 125I-radiolabeled growth factor to the cell surface. These assays revealed that EGF receptors are found on five of six non-SCCL cell lines and are not found on any of the SCCL lines. In contract, NGF binding was detected at low levels on three of eight SCCL lines and on all three SCCL converters but was not observed for non-SCCL lines. Thus, SCCL and SCCL converter cell lines are distinguished from non-SCCL by the pattern of membrane receptors for EGF and NGF. Such differences may ultimately prove useful as biological markers for the different histological types of lung cancer. Moreover, the majority of SCCL cells and all of the non-SCCL cells tested were found to produce diffusible growth factors which can stimulate soft agar growth of nontransformed normal rat kidney fibroblasts. Although some correlation between soft agar growth factor production and the absence of EGF receptors may exist for SCCL cells, the production of transforming growth factors appears to be a general property of human lung cancer cells in vitro and is independent of EGF receptor expression.

Topics: Agar; Cell Line; Cell Membrane; Epidermal Growth Factor; Growth Substances; Humans; Lung Neoplasms; Nerve Growth Factors; Peptides; Protein Binding

The effect of mechanical and enzymatic disaggregation on human malignant melanoma, soft-tissue sarcoma and lung carcinoma colony growth in soft agar was studied. The enzymatic disaggregation was advantageous in most cases of melanoma and sarcoma, giving a larger number of colonies and increasing the probability of achieving growth in soft agar. Enzymatically treated pulmonary carcinoma cell populations had lower clonogeneic potential, especially in the case of anaplastic carcinomas. Morphological studies showed that the cells growing in soft-agar colonies had the same characteristics as those of the original tumor. A linear relationship was obtained between the number of enzymatically and mechanically treated tumor cells plated and the number of colonies. Delayed plating decreased the number of colonies.

Topics: Agar; Carcinoma; Cell Aggregation; Clone Cells; Cytological Techniques; Humans; Lung Neoplasms; Melanoma; Microbial Collagenase; Sarcoma; Soft Tissue Neoplasms; Time Factors

The pattern of in vitro anchorage-independent growth of tumor cells from the murine UV-2237 fibrosarcoma correlated with their ability to produce experimental metastasis in vivo. When seeded into 0.3% Noble agar semisolid medium, cells of metastatic clones developed into larger tumor colonies at a faster rate than did cells of clones with low metastatic potential. Furthermore, when tumor cells were plated into 0.6% Noble agar, colony development by cells of low metastatic potential clones was almost completely restricted. Tumor cells from the heterogeneous parent UV-2237 fibrosarcoma were plated into dishes containing 0.6% agar semisolid medium. In separate experiments, 16 colonies were isolated 2 weeks thereafter and were established as individual cell lines in monolayer cultures. All of these cell lines produced experimental metastases as determined by in vivo lung colony assay. The data suggest that anchorage-independent growth of UV-2237 tumor cells in 0.6% Noble agar semisolid medium is selective and permits the isolation of metastatic subpopulations.

Topics: Agar; Animals; Cell Adhesion; Cell Division; Cells, Cultured; Culture Media; Fibrosarcoma; Lung Neoplasms; Mice; Neoplasm Metastasis; Sarcoma, Experimental

Inhibition of leukocyte migration in agarose-agar was used as a probe for tumor-associated antigen in 3-M KCl solubilized extracts of gastric, colon, and lung cancers from humans. Twelve of 40 (30%) leukocyte preparations from gastric cancer patients, 10 of 21 (48%) from colon cancer patients, and 7 of 14 (50%) from lung cancer patients were inhibited by their respective histologically homologus cancer extract. However, among 75 preparations from various cancer patients, leukocytes from only 2 gastric cancer patients were inhibited by paired normal gastric tissue extracts. Only 2 of 68 preparations from normal individuals and none of 67 preparations from patients with nonmalignant diseases, such as gastric peptic ulcer, gastritis, colon polyposis, colitis, pulmonary tuberculosis, chronic bronchitis, and sarcoidosis, were inhibited by cancer extracts. These findings suggest the presence in KCl extracts of gastric cancer of presumed tumor-associated antigen(s) that is antigenically distinct from that of either colon or lung cancer.

Topics: Agar; Antigens, Neoplasm; Cell Migration Inhibition; Colonic Neoplasms; Female; Humans; Leukocytes; Lung Neoplasms; Male; Potassium Chloride; Stomach Neoplasms; Tissue Extracts

A soft agar colony assay has been developed for the B16 mouse melanoma and the Lewis lung tumour. The special features of the technique are the use of a gas phase with 5% O2 instead of air and the addition of rat red blood cells. Single cell suspensions are prepared by trypsinization from the solid tumour and the cells are plated out in 0-3% agar over a layer of 0-5% agar in 30-mm Petri dishes. After 8 to 15 days' incubation in 5% O2, colonies of more than 50 cells are produced. Plating efficiencies of between 30 and 50% are usually obtained. The addition of up to 10(4) heavily irradiated tumour cells gives some further improvement in plating efficiency for the B16 melanoma but not for the Lewis lung tumour. Applications of the technique to measure cell survival in the two tumours after treatment with cytotoxic drugs and radiation are reported. The scatter of experimental points is relatively small, and in comparative experiments good agreement has been obtained with results using in vivo assay techniques.

Topics: Agar; Animals; Cell Count; Cell Survival; Clone Cells; Cyclophosphamide; Cytological Techniques; Erythrocytes; Gamma Rays; Lung Neoplasms; Melanoma; Mice; Neoplasms, Experimental; Oxygen; Rats; Time Factors

The frequency of M-components was studied by agar gel electrophoresis in sera from 807 patients, 467 (57%) females and 340 (43%) males with histologically proven solid malignant neoplasms.M-components were found in the sera of 40 male and 20 female patients. Apart from two known cases of multiple myeloma and one case of Waldenström's macroglobulinaemia, none of the patients were found to be suffering from these diseases. The frequency of M-components increased with age, and this was more evident in males. Twenty-two of 60 patients with M-components did not exhibit abnormalities on immunoelectrophoresis. Of the 35 remaining patients, 27 had an abnormal component of the IgG class, 6 of the IgA and 2 of the IgM class. M-components were found in the sera of patients with a wide variety of neoplasms. There appeared to be no evidence of an increased frequency of M-components in the sera of patients with solid malignant neoplasms compared with normal adult population.

Topics: Adolescent; Adult; Agar; Aged; Child; Electrophoresis; Female; Gastrointestinal Neoplasms; Genital Neoplasms, Female; Humans; Immunoelectrophoresis; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Lung Neoplasms; Male; Middle Aged; Multiple Myeloma; Neoplasms; Urinary Bladder Neoplasms; Waldenstrom Macroglobulinemia

Topics: Agar; Bacteria; Bacteriological Techniques; Blood; Bronchi; Bronchoscopy; Chronic Disease; Cough; Culture Media; Female; Humans; Lung Diseases; Lung Neoplasms; Male; Prospective Studies; Sex Factors; Smoking; Sputum

Topics: Agar; Amylases; Electrophoresis; Humans; Isoenzymes; Lung Neoplasms

Topics: Agar; Aminohydrolases; Diagnosis, Differential; Electrophoresis; Gels; Humans; Lung Neoplasms; Methods; Tuberculosis, Pulmonary

Topics: Agar; Aged; Blood Protein Electrophoresis; Breast Neoplasms; Carcinoma, Bronchogenic; Colonic Neoplasms; Esophageal Neoplasms; Female; gamma-Globulins; Humans; Kidney Neoplasms; Lung Neoplasms; Middle Aged; Neoplasm Metastasis; Neoplasms; Stomach Neoplasms

Topics: Agar; Histological Techniques; Humans; Lung Abscess; Lung Diseases; Lung Neoplasms