agar and Adenocarcinoma

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 evaluation of new drug treatments and combination treatments for gliomas and other cancers requires a robust means to interrogate wide dose ranges and varying times of drug exposure without stain-inactivation of the cells (colonies). To this end, we developed a 3-dimensional (3D) colony formation assay that makes use of GelCount technology, a new cell colony counter for gels and soft agars. We used U251MG, SNB19, and LNZ308 glioma cell lines and MiaPaCa pancreas adenocarcinoma and SW480 colon adenocarcinoma cell lines. Colonies were grown in a two-tiered agarose that had 0.7% agarose on the bottom and 0.3% agarose on top. We then studied the effects of DFMO, carboplatin, and SAHA over a 3-log dose range and over multiple days of drug exposure. Using GelCount we approximated the area under the curve (AUC) of colony volumes as the sum of colony volumes (microm2xOD) in each plate to calculate IC50 values. Adenocarcinoma colonies were recognized by GelCount scanning at 3-4 days, while it took 6-7 days to detect glioma colonies. The growth rate of MiaPaCa and SW480 cells was rapid, with 100 colonies counted in 5-6 days; glioma cells grew more slowly, with 100 colonies counted in 9-10 days. Reliable log dose versus AUC curves were observed for all drugs studied. In conclusion, the GelCount method that we describe is more quantitative than traditional colony assays and allows precise study of drug effects with respect to both dose and time of exposure using fewer culture plates.

Topics: Adenocarcinoma; Agar; Antineoplastic Agents; Area Under Curve; Carboplatin; Cell Line, Tumor; Cell Proliferation; Colony-Forming Units Assay; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Humans; Inhibitory Concentration 50; Sepharose

The epidermal growth factor receptor (EGFR) autocrine signaling pathway is involved in cancer development and progression. EGFR inhibitors such as C225 (cetuximab), a chimeric human-mouse anti-EGFR monoclonal antibody, and ZD1839 (gefitinib), a small molecule EGFR-selective tyrosine kinase inhibitor, are in advanced clinical development. The potential emergence of cancer cell resistance in EGFR-expressing cancers treated with EGFR inhibitors could determine lack of activity of these drugs in some cancer patients. Vascular endothelial growth factor (VEGF) is secreted by cancer cells and plays a key role in the regulation of tumor-induced endothelial cell proliferation and permeability. ZD6474 is a small molecule VEGF flk-1/KDR (VEGFR-2) tyrosine kinase inhibitor that also demonstrates inhibitory activity against EGFR tyrosine kinase.. The antitumor activity of ZD1839, C225, and ZD6474 was tested in athymic mice bearing human GEO colon cancer xenografts. GEO cell lines resistant to EGFR inhibitors were established from GEO xenografts growing in mice treated chronically with ZD1839 or C225. Expression of EGFR was evaluated by flow cytometry. Expression of various proteins involved in intracellular cell signaling was assessed by Western blotting. Tumor growth data were evaluated for statistical significance using the Student's t test. All Ps were two-sided.. Although chronic administration of optimal doses of C225 or ZD1839 efficiently blocked GEO tumor growth in the majority of mice, tumors slowly started to grow within 80-90 days, despite continuous treatment. In contrast, continuous treatment of mice bearing established GEO xenografts with ZD6474 resulted in efficient tumor growth inhibition for the entire duration of dosing (up to 150 days). ZD6474 activity was also determined in mice pretreated with ZD1839 or C225. When GEO growth was apparent after 4 weeks of treatment with EGFR inhibitors, mice were either re-treated with EGFR inhibitors or treated with ZD6474. GEO tumor growth was blocked only in mice treated with ZD6474, whereas tumor progression was observed in mice re-treated with C225 or ZD1839. GEO tumors growing during treatment with C225 or with ZD1839 were established as cell lines (GEO-C225-RES and GEO-ZD1839-RES, respectively). Cell membrane-associated EGFR expression was only slightly reduced in these cell lines compared with parental GEO cells. Western blotting revealed no major change in the expression of the EGFR ligand transforming growth factor alpha of bcl-2, bcl-xL, p53, p27, MDM-2, akt, activated phospho-akt, or mitogen-activated protein kinase. However, both GEO-C225-RES and GEO-ZD1839-RES cells exhibited a 5-10-fold increase in activated phospho-mitogen-activated protein kinase and in the expression of cyclooxygenase-2 and of VEGF compared with GEO cells. GEO-C225-RES and GEO-ZD1839-RES growth as xenografts in nude mice was not significantly affected by treatment with either C225 or ZD1839 but was efficiently inhibited by ZD6474.. Long-term treatment of GEO xenografts with selective EGFR inhibitors results in the development of EGFR inhibitor-resistant cancer cells. Growth of EGFR inhibitor-resistant tumors can be inhibited by ZD6474. These data indicate that inhibition of VEGF signaling has potential as an anticancer strategy, even in tumors that are resistant to EGF inhibitors.

Topics: Adenocarcinoma; Agar; Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Antineoplastic Agents; Blotting, Western; Cell Division; Cell Line, Tumor; Cell Membrane; Cetuximab; Colonic Neoplasms; Drug Resistance, Neoplasm; ErbB Receptors; Female; Flow Cytometry; Humans; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Piperidines; Precipitin Tests; Quinazolines; Receptors, Vascular Endothelial Growth Factor; Time Factors

Cancer of unknown primary origin is the eighth most common form of malignancy and accounts for up to 10% of all neoplasms diagnosed. It is a set of heterogenous tumors with widely varying sensitivity to systemic chemotherapy. Over the past years progress has been made in identifying subsets of patients that can be effectively treated with chemotherapy and may achieve long-term survival even with metastatic disease. However, the large majority of cancers of unknown origin still is resistant to chemotherapy. In an attempt to identify conventional and investigational new agents with possible activity against cancers of unknown primary, we have retrospectively analyzed the results of chemosensitivity testing in a soft agar cloning system in vitro and have compared these data with published clinical trials. Between 1978 and 1993, a total of 19584 tumor specimens were studied using a variety of investigational or established antitumor agents. Of these, 615 (3.1%) were tumors of unknown origin and confirmed on pathology review. The largest histologic subgroup was adenocarcinoma (332, 54%). Sufficient numbers of cells for in vitro testing were obtained from 313 tumor specimens (94.3%). Of 278 agents tested in adenocarcinoma of unknown origin, borderline activity (< 20% in vitro response) was noted for 5-FU, doxorubicin, bleomycin, mitoxantrone, mitomycin-C, cisplatin, and etoposide. In vitro response rates of > or = 20% were observed for actinomycin-D, BCNU, melphalan, methotrexate, taxol, and vinblastine. In addition, several investigational agents including fludarabine, amira235, bisantrene, Dupont840, echinomycin, tiazofurin, LY104208 (vinzolidine), intoplicine, and topotecan had activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenocarcinoma; Agar; Drug Screening Assays, Antitumor; Humans; Neoplasms, Unknown Primary; Retrospective Studies; Tumor Stem Cell Assay

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

The three-dimensional growth of cultured tumor cell lines (HT29, a colon adenocarcinoma cell line; M21, a melanoma cell line; KB, a nasopharyngeal carcinoma cell line) has been investigated in an agar culture system with a fibrin matrix in vitro. The tumor cells developed to tumors 3 x 3 mm in diameter after 10 days in culture in vitro. This size was large enough to allow histologic examination. The tumor cells located in the surface area of the three-dimensional tumor seemed to grow well. However, the tumor cells in the center degenerated or did not proliferate, indicating a lack of nutrition and/or anoxia in the center. The histologic comparison between the xenografted tumors on nude mice and the three-dimensional tumors in vitro suggests that the structures of the three-dimensional tumors were comparable, especially in the surface area, to the xenografted tumors. Furthermore, the antitumor effect of mitomycin C on the three-dimensional tumors was found to be dose-dependent.

Topics: Adenocarcinoma; Agar; Animals; Cell Division; Cell Hypoxia; Colonic Neoplasms; Culture Techniques; Extracellular Matrix; Fibrin; Humans; KB Cells; Melanoma; Mice; Mice, Nude; Mitomycin; Transplantation, Heterologous; Tumor Cells, Cultured

Morphofunctional peculiarities of tumor cells from 15 endometrial adenocarcinomas and 2 ovarian tumors have been investigated at the ultrastructural level. These cells could develop two types of colonies in soft agar: those with histotypical differentiation (numerous microvilli, well developed tight junctions, desmosomes, secretory granules), and those without it (absence of epithelial features, ability of tumor cells to produce filamentous extracellular matrix and striated collagen fibrils which are characteristic of fibroblastic cells). The addition of progesterone and tamoxifen to cell cultures resulted in rising the level of cell differentiation in the colonies. The fact that endometrial and ovarian cancer cells can express the properties specific of connective tissue cells may suggest a multipotention of the Mullerian epithelium derivatives to shed light on the histogenesis of the mixed Mullerian tumors of uterus.

Topics: Adenocarcinoma; Agar; Cell Transformation, Neoplastic; Culture Media; Cystadenocarcinoma; Endometrial Neoplasms; Female; Granulosa Cell Tumor; Humans; Methotrexate; Microscopy, Electron; Morphogenesis; Ovarian Neoplasms; Progesterone; Tamoxifen; Tumor Cells, Cultured

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

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

Topics: Adenocarcinoma; Agar; Antineoplastic Agents; Cells, Cultured; Colonic Neoplasms; Colorimetry; Culture Media; Humans

Radiation survival curves have been generated for 3 human tumor cell lines as a means of comparing and evaluating the validity of human tumor soft-agar clonogenic assays. The assays investigated were the Hamburger-Salmon, Courtenay-Mills, Courtenay-Mills plus additions, soft agar (no additions), and soft agar plus additions. The additions were formulated to supplement the media used in soft agar assays of primary ovarian and cervical carcinoma specimens. Supplementing the media with additions led to a 2- to 3-fold increase in PE of CaSki cells but had no effect on the PEs of ME180 and OWI cells. Radiation survival curves were similar in all assays for CaSki and OWI but differed for ME180 cells. For ME180 cells, the Courtenay-Mills and soft agar assays plus additions produced the most radioresistant curves (Do = 2.2 Gy); the cells were more responsive when assayed by the Hamburger-Salmon method (Do = 1.5 Gy), and the soft agar and Courtenay-Mills assays gave the most radiosensitive curves (Do = 1.2 Gy). These results demonstrate that the PE of human tumor cell lines may be increased with no effect on radiation survival; radiation survival may be altered without changes in PE and neither may be altered by applying modifications and supplements to existing clonogenic assays.

Topics: Adenocarcinoma; Agar; Carcinoma, Squamous Cell; Cell Line; Cell Survival; Colony-Forming Units Assay; Female; Humans; Neoplasms; Ovarian Neoplasms; Tumor Stem Cell Assay; Uterine Cervical Neoplasms

Topics: Adenocarcinoma; Agar; Cells, Cultured; Colony-Forming Units Assay; Culture Media; Cystadenoma; Female; Humans; In Vitro Techniques; Ovarian Neoplasms; Tumor Stem Cell Assay

A human epithelial-derived cell line, SW-13, releases a soluble substance that functions as an autocrine growth factor. SW-13 cells, derived from a human adenocarcinoma of the adrenal cortex, form a few small colonies when suspended in soft agar at low densities. The number of colonies increased significantly when either viable SW-13 cells or serum-free medium conditioned by SW-13 cells (CM) was added to agar underlayers. CM increased colony formation in a dose-dependent fashion. Clonal growth at low cell densities was dependent on the presence of both horse serum and SW-13 CM. Neither activity alone was capable of sustaining growth. Even when cells were plated at high densities CM could not substitute for serum, but could reduce the threshold serum concentration. The results suggest that autocrine and serum-derived factors act in concert to maintain clonal growth of epithelial tumor cells in soft agar.

Topics: Adenocarcinoma; Adrenal Gland Neoplasms; Agar; Cell Division; Cell Line; Culture Media; Epithelium; Growth Substances; Humans

Soft agar cultures of human prostatic carcinoma cells obtained from prostates with needle biopsy and metastatic lymph nodes were examined. Colony formation were observed in 2 of the 4 needle biopsy specimens and in 7 of the 10 lymphatic nodes specimens. The plating efficiency was 0.04 to 0.43%. The effect of testosterone on the clonal growth of these cells in soft agar culture was studied. In half of the cases, colony formation was increased by the addition of testosterone. Whereas in the other cases, colony formation was not influenced by the testosterone. Clinically, most of the former cases responded well to the anti-androgen therapy. Neither EGF nor hydrocortisone showed any effect on the colony formation of the prostatic cancer cells in soft agar. These results suggest that primary culture of human prostatic cancer is possible in soft agar medium, and that androgen dependency of these cells can be examined in vitro.

Topics: Adenocarcinoma; Agar; Aged; Cells, Cultured; Colony-Forming Units Assay; Culture Media; Humans; Male; Middle Aged; Prostatic Neoplasms; Testosterone; Tumor Stem Cell Assay

One hundred and thirty three specimens from mammary and ovarian adenocarcinoma and from melanoma were cultured according to an agar/agar clonogenic assay. Melanoma and ovarian cancers exhibited a 70 per cent rate of success for culture; 50 per cent of the mammary adenocarcinomas were successfully cultured. Fifty-nine ovarian cancers were cultured in order to test the in vitro effectiveness of Cisplatinum and Adriamycin. Thirty percent of cultured tumors gave rise to relevant chemograms. The chemoresistance measured in vitro was correlated to the ineffectiveness of the patient's treatment. In contrast, we were unable to predict chemosensitivity. Taking into account the technical difficulties encountered in these assays, human tumor clonogenic assays cannot at present be proposed as a routine procedure in the prediction of the effectiveness of chemotherapeutic treatments. Nevertheless, they must be developed in order to determine the spectrum of activity of new antineoplastic agents on various human tumors.

Topics: Adenocarcinoma; Agar; Antineoplastic Agents; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Cisplatin; Colony-Forming Units Assay; Cyclophosphamide; Doxorubicin; Drug Resistance; Female; Humans; Melanoma; Melphalan; Ovarian Neoplasms; Tumor Stem Cell Assay

Two hundred six different samples of human renal carcinoma were tested for in vitro chemotherapy sensitivity using a soft agar colony formation assay similar to that originally described by Salmon and colleagues. Eighty of 159 (50 per cent) evaluable tumor tests showed colony formation in vitro and gave clinical drug sensitivity information. Two-thirds of tumors were resistant to all drugs tested, despite a median number of drugs tested per tumor of 14.5. Five tumors (6 per cent) were remarkably sensitive to numerous anticancer drugs in vitro. The most active drugs found in vitro were teniposide, actinomycin D, bleomycin, hydroxyurea, mitoguanazone dihydrochloride, mitomycin C and L-alanosine. Fourteen other drugs tested showed low in vitro cytotoxicity.

Topics: Adenocarcinoma; Agar; Antineoplastic Agents; Bleomycin; Cells, Cultured; Dactinomycin; Drug Evaluation; Humans; Hydroxyurea; Kidney Neoplasms; Mitoguazone; Nephrectomy; Teniposide

We have systematically analyzed the proliferative properties of the clonogenic cells of the R3327 transplantable rat prostate adenocarcinoma (colony forming units, prostatic adenocarcinoma, CFU-PA). Pronase was determined to be more effective than either trypsin or collagenase in obtaining the largest number of viable cells from a solid R3327 tumor. Digestion with 2.5 mg/ml yielded a maximum of 5.6 X 10(4) CFU-PA/g of tumor tissue, with higher concentrations resulting in s substantially lower fraction of CFU-PA while producing larger overall cell yields. Bacto-agar at 0.35% supported the growth of the largest number of CFU-PA and was sharply concentration dependent with concentrations greater than 0.4% completely suppressing CFU-PA growth. The addition of conditioned medium (CM) from R3327 liquid cell cultures to agar cultures resulted in a specific twofold increase in CFU-PA/10(4) cells, whereas CM from R3327A cells was less effective and CM from rat skin fibroblasts least stimulatory. The addition of washed rat red blood cells (RBC) either within the agar culture itself or in an overlayer was highly stimulatory, resulting in as much as a fivefold increase in CFU-PA to 6 to 8 x 10(4)/g.

Topics: Adenocarcinoma; Agar; Animals; Cell Count; Cell Division; Cell Line; Erythrocytes; Female; Male; Mercaptoethanol; Neoplasms, Experimental; Peptide Hydrolases; Pronase; Prostatic Neoplasms; Rats; Rats, Inbred Strains; Thrombin

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

Topics: Adenocarcinoma; Agar; Aged; Antineoplastic Agents; Colony-Forming Units Assay; Culture Media; Drug Evaluation, Preclinical; Female; Humans; Kidney Neoplasms; Male; Middle Aged; Tumor Stem Cell Assay

A well-differentiated colorectal tumor T 219 which grows as a xenograft in athymic mice (human-tumor-nude-mouse system) and forms colonies in culture (soft agar colony-formation assay) has been used to test the correlation between the above two methods of exposure of human tumor cells to antineoplastic agents. In in vitro studies, two protocols were used: 1 h drug exposure and continuous drug exposure. In the 1 h drug exposure experiments six drugs, doxorubicin (DX), 4'-deoxydoxorubicin (deoDX), 4'epidoxorubicin(epiDX), 4'-O-methyldoxorubicin (O-DX), N-trifluoroacetyldoxorubicin-14-valerate (AD-32) and 5-fluorouracil (FUra) were studied, while in continuous drug exposure experiments four of the above drugs (DX, deoDX, epiDX, O-DX) were studied. The survival of the tumor clonogenic cells (HC219) was determined by counting the number of colonies formed during 13-14 days of incubation and dose-response curves were obtained. In in vivo studies, the mice were treated with all of the drugs used in in vitro 1 h drug exposure experiments (DX, deoDX, epiDX, O-DX, AD-32 and FUra). To quantitate the chemotherapeutic effectiveness of the drugs, T/C% (relative tumor volume of treated group as percentage of the control group) values were calculated each time the tumors were measured. The experimental data suggest that in vitro 1 h drug exposure results are in good agreement with the in vivo results, while the continuous drug exposure results do not agree with the in vivo data. The most active drug in in vivo studies, deoDX, was found to be the most active drug in the in vitro 1 h drug exposure experiments as well. However, in continuous drug exposure experiments, O-DX, not deoDX, was found to be the most active drug. Activities of the other drugs tested also differed from their respective activities in in vivo studies. Although the relative effectiveness of various drugs can be compared by determining molar concentrations of the drugs producing 50% inhibition of colonies (ID50) the expression, PEI = LD10/ID50 X 1000, which takes into consideration toxicity of the drugs, is probably a better indicator of the in vitro drug activity. The results suggest that soft agar colony-formation assay (with established cell lines from the same tumor) may be used for the prediction of in vivo activity of potential antineoplastic agents against human tumor xenografts in nude mice.

Topics: Adenocarcinoma; Agar; Animals; Antineoplastic Agents; Cell Line; Cell Survival; Clone Cells; Colonic Neoplasms; Dose-Response Relationship, Drug; Evaluation Studies as Topic; Female; Humans; Mice; Mice, Nude; Neoplasm Transplantation

Dispersed tumor cells from primary human renal adenocarcinomas showed typical clonogenic growth in soft agar in seven of 31 instances. In vitro chemotherapy sensitivity testing was performed successfully for five of the seven clonogenic tumors. Extensive inherent resistance to the cytotoxic effects of many standard and experimental chemotherapeutic agents were observed. Occasionally, a single drug showed marked cytotoxicity. Similar results were obtained with soft agar clonogenic chemotherapeutic drug sensitivity assays performed on cells from human renal carcinoma xenografts. These in vitro results agree with clinical experience in the chemotherapy of advanced renal carcinoma. It remains to be determined if the sensitivity or resistance to chemotherapeutic drugs of human renal carcinomas noted in vitro reflects the characteristics of the tumors in vivo. Our results show that it is feasible to evaluate the soft agar assay as a method for selecting drugs for individual patients with renal carcinoma, a disease for which no chemotherapeutic treatment can be recommended at present.

Topics: Adenocarcinoma; Agar; Animals; Antineoplastic Agents; Cells, Cultured; Colony-Forming Units Assay; Drug Resistance; Humans; Kidney Neoplasms; Male; Mice; Mice, Nude; Neoplasm Transplantation

Topics: Adenocarcinoma; Adenocarcinoma, Mucinous; Agar; Cell Division; Chromosome Aberrations; Cisplatin; Clone Cells; Cystadenocarcinoma; Drug Resistance; Endometriosis; Female; Humans; Methods; Neoplasms, Experimental; Ovarian Neoplasms; Phagocytes

Topics: Adenocarcinoma; Adenocarcinoma, Scirrhous; Adolescent; Adult; Agar; Aged; Carcinoma; Chromatography; Colonic Neoplasms; Gallbladder Neoplasms; Hemangiosarcoma; Humans; Liver Neoplasms; Lymphoma, Non-Hodgkin; Middle Aged; Neoplasm Proteins; Neoplasm Recurrence, Local; Pancreatic Neoplasms; Protein Denaturation; Retroperitoneal Neoplasms; Stomach Neoplasms

Epithelial-like cells originating from 10-day and 8-week old BD rats were treated in vitro with dimethylnitrosamine (DMN) and N-methyl-N'-nitro-nitrosoguanidine (MNNG). Morphologically, no differences were seen between treated and untreated cells up to the time when the cells were transplanted into syngeneic hosts. However, the treated cells grew in soft agar and once injected subcutaneously or intraperitoneally into newborn rats, developed tumours after a latent period of 9-12 weeks. The tumours obtained with DMN-treated cells were well differentiated adenocarcinomata, whereas carcinosarcomata were observed with the MNNG-treated cells.

Topics: Adenocarcinoma; Agar; Animals; Carcinoma; Cell Line; Epithelial Cells; Injections, Intraperitoneal; Injections, Subcutaneous; Liver Neoplasms; Neoplasm Transplantation; Nitrosamines; Nitrosoguanidines; Rats

A herpes-type virus was induced in explants of "summer-phase" Lucké tumor maintained on agar slants at 7.5 C for 13 to 14 weeks.

Topics: Adenocarcinoma; Agar; Animals; Anura; Cell Nucleus; Cold Temperature; Culture Media; Culture Techniques; Cytoplasm; Herpesviridae; Inclusion Bodies, Viral; Kidney Neoplasms; Microscopy, Electron; Time Factors

Topics: Adenocarcinoma; Adult; Agar; Animals; Antineoplastic Agents; Anus Neoplasms; Breast Neoplasms; Carcinoma, Squamous Cell; DNA; Dogs; Humans; Leucine; Leukocytes; Liver; Mice; Middle Aged; Neoplasms; Neoplasms, Experimental; Parotid Neoplasms; Proteins; Radioisotopes; Rectal Neoplasms; RNA; Sulfhydryl Compounds; Thoracic Neoplasms; Thymidine; Uridine

Topics: Adenocarcinoma; Adolescent; Adult; Agar; Aged; Blood Protein Electrophoresis; Endometriosis; Female; Humans; Isoenzymes; L-Lactate Dehydrogenase; Leiomyoma; Leiomyosarcoma; Middle Aged; Polyps; Spectrophotometry; Uterine Neoplasms