cytochalasin-b and Lung-Neoplasms

Cytochalasin B is a potentially novel microfilament-directed chemotherapeutic agent that prevents actin polymerization, thereby inhibiting cytokinesis. Although cytochalasin B has been extensively studied in vitro, only limited data are available to assess its in vivo potential. Cytochalasin B was administered to Balb/c mice challenged i.d. with M109 murine lung carcinoma to determine whether the agent could affect an established i.d. tumor when the compound is administered s.c. in the region of the i.d. tumor, but not in direct contact with it. Cytochalasin B was also administered either i.p. or s.c. at a distant site or i.v. to determine whether it could affect the long-term development of an established i.d. tumor. Cytochalasin B was then liposome encapsulated to determine whether the maximum tolerated dose (MTD) of the compound could be increased, while reducing immunosuppression that we have previously characterized. Liposomal cytochalasin B was also administered to mice challenged i.d. with M109 lung carcinoma to assess its chemotherapeutic efficacy. The results can be summarized as follows: 1) cytochalasin B substantially delayed the growth of i.d. M109 tumor nodules, inhibited metastatic progression in surrounding tissues, and produced long-term cures in treated mice; 2) liposomal cytochalasin B increased the i.p. MTD by more than 3-fold, produced a different distribution in tissue concentrations, and displayed antitumor effects against M109 lung carcinoma similar to non-encapsulated cytochalasin B. These data show that cytochalasin B exploits unique chemotherapeutic mechanisms and is an effective antineoplastic agent in vivo in pre-clinical models, either in bolus form or after liposome encapsulation.

Topics: Animals; Antineoplastic Agents; Cytochalasin B; Disease Progression; Dose-Response Relationship, Drug; Drug Administration Routes; Liposomes; Lung Neoplasms; Maximum Tolerated Dose; Mice; Mice, Inbred BALB C

Background Despite inherent differences between the cytoskeletal networks of malignant and normal cells, and the clinical antineoplastic activity of microtubule-directed agents, there has yet to be a microfilament-directed agent approved for clinical use. One of the most studied microfilament-directed agents has been cytochalasin B, a mycogenic toxin known to disrupt the formation of actin polymers. Therefore, this study sought to expand on our previous work with the microfilament-directed agent, along with other less studied cytochalasin congeners. Materials and Methods We determined whether cytochalasin B exerted significant cytotoxic effects in vitro on adherent M109 lung carcinoma and B16BL6 and B16F10 murine melanomas, or on suspension P388/ADR murine leukemia cells. We also examined whether cytochalasin B, its reduced congener 21, 22-dihydrocytochalasin B (DiHCB), or cytochalasin D could synergize with doxorubicin (ADR) against ADR-resistant P388/ADR leukemia cells, and produce significant cytotoxicity in vitro. For in vivo characterization, cytochalasins B and D were administered intraperitoneally (i.p.) to Balb/c mice challenged with drug sensitive P388-S or multidrug resistant P388/ADR leukemias. Results Cytochalasin B demonstrated higher cytotoxicity against adherent lung carcinoma and melanoma cells than against suspension P388/ADR leukemia cells, as assessed by comparative effects on cell growth, and IC₅₀ and IC₈₀ values. Isobolographic analysis indicated that both cytochalasin B and DiHCB demonstrate considerable drug synergy with ADR against ADR-resistant P388/ADR leukemia, while cytochalasin D exhibits only additivity with ADR against the same cell line. In vivo, cytochalasins B and D substantially increased the life expectancy of mice challenged with P388/S and P388/ADR leukemias, and in some cases, produced long-term survival. Conclusion Taken together, it appears that cytochalasins have unique antineoplastic activity that could potentiate a novel class of chemotherapeutic agents.

Topics: Animals; Antineoplastic Agents; Cell Survival; Cytochalasin B; Cytochalasin D; Cytochalasins; Doxorubicin; Drug Synergism; Leukemia P388; Lung Neoplasms; Melanoma; Mice; Mice, Inbred BALB C; Neoplasms; Tumor Cells, Cultured

Several studies have shown that curcumin can induce apoptosis and inhibit growth in human A549 lung adenocarcinoma cells. However, the mechanism is not completely understood yet. In the present study, we investigated the in vitro effect of curcumin on cell viability, apoptosis and disorganization of the actin cytoskeleton in A549 cells. Our results showed that curcumin significantly inhibited the viability of A549 cells in a dose- and time-dependent manner by induced apoptosis. The apoptotic process was associated with a disorganization of the architecture of actin microfilaments and a decrease in the levels of F-actin. DMSO-treated control cells exhibited a well-defined F-actin network that was mainly organized into stress fibers. The actin fibers in cells treated with curcumin or the positive control drug cytochalasin B were disorganized, disassembled, or disrupted, however, the disorganization of actin fibers and apoptosis could be prevented by phalloidin, an F-actin stabilizing compound. Thus, these results demonstrated that actin filament disorganization might play a central role in the curcumin-induced apoptosis of A549 cells.

Topics: Actin Cytoskeleton; Actins; Adenocarcinoma; Antineoplastic Agents, Phytogenic; Apoptosis; Cell Line, Tumor; Cell Survival; Curcuma; Curcumin; Cytochalasin B; Cytoskeleton; Dose-Response Relationship, Drug; Humans; Lung Neoplasms; Phalloidine; Phytotherapy; Plant Extracts; Poisons

In this study, the endocytosis and the internalization mechanism of aminosilane-coated Fe(3)O(4) nanoparticles into human lung cancer cell line SPC-A1 was studied compared with human lung cell line WI-38 in vitro. The particle endocytosis behavior was studied by using Transmission Electron Microscope (TEM) and Coupled Plasma-Atomic Emission Spectrometry (ICP-AES). It was found that aminosilane-coated Fe(3)O(4) nanoparticles could be greatly taken up by SPC-A1 human cancer cells (202 pg iron/cell) but not by WI-38 human lung cells (13 pg iron/cell). The particles could be retained in SPC-A1 cells over a number of generations in vitro. Different endocytosis was observed by TEM after SPC-A1 cells were treated with different temperature or with/without Cytochalasin B (Inhibitor of phagocytosis) at 37 degrees C. No nanoparticles were taken up by SPC-A1 after the endocytosis inhibited in low temperature. Restoring the endocytosis activity at 37 degrees C, the process of nanoparticles from coated pit to endosomes and lysosomes was observed by TEM. Endocytosis activity was effectively inhibited by the presence of Cytochalasin B at 37 degrees C, while a lot of nanoparticles were uptaken to the cytoplasm of SPC-A1 cells in the control group. Our results suggest that the process of endocytosis of aminosilane-coated Fe(3)O(4) nanoparticles can efficiently takes place in lung cancer cells and nanoparticles can be kept in cancer cells for generations. Phagocytosis may be involved in the internalization process of aminosilane-coated Fe(3)O(4) nanoparticles.

Topics: Cell Line, Tumor; Coated Materials, Biocompatible; Cytochalasin B; Endocytosis; Ferric Compounds; Humans; Iron; Lung Neoplasms; Magnetics; Metal Nanoparticles; Silanes; Temperature; Time Factors

Survivin and securin proteins are overexpressed in most cancer cells that have been shown to regulate mitotic progression. In this study, we investigated the roles of survivin and securin on cytochalasin B, a cytokinesis blocker mediating the cytotoxicity and cell growth inhibition in human cancer cells. The human lung carcinoma cell lines A549 and H1299 highly expressed survivin proteins in mitosis and concentrated on the midbodies during cytokinesis. Cytochalasin B significantly decreased cell survival, inhibited cell growth, increased the levels of G(2)/M fractions, and induced binuclei formation in lung carcinoma cells; however, the survivin proteins were concentration-dependently increased by 1 to 5 mug/ml cytochalasin B for 24 h. It is noteworthy that the expression of securin proteins was decreased in cytochalasin B-treated lung carcinoma cells. Transfection of 20 to 40 nM survivin siRNA for 48 h significantly induced the formation of multiple nuclei and apoptosis but decreased the levels of survivin and securin proteins in A549 cells. Cotreatment with survivin small interfering RNA (siRNA) and cytochalasin B increased the cytotoxicity and cell growth inhibition. In addition, the securin-null colorectal carcinoma cells were more susceptible to the cytotoxicity after cytochalasin B and survivin siRNA treatments than the securin-wild-type cells. As a whole, our results indicate that the inhibition of survivin and securin protein expression may increase the cell death and growth inhibition after cytochalasin B treatment in human cancer cells.

Topics: Cell Cycle; Cell Death; Cell Division; Cell Line, Tumor; Colorectal Neoplasms; Cytochalasin B; Fluorescent Antibody Technique; Humans; Inhibitor of Apoptosis Proteins; Lung Neoplasms; Microscopy, Confocal; Microtubule-Associated Proteins; Neoplasm Proteins; RNA, Small Interfering; Securin; Survivin

We studied micronucleus frequencies in former German uranium miners of the Wismut SDAG (Sowjetisch-Deutsche Aktiengesellschaft). Various other groups were analyzed for comparison (individuals with lung tumors or lung fibrosis, controls). We had shown previously that micronucleus frequencies were not different among the various groups. Differences were observed, however, when centromere-positive and -negative micronuclei were distinguished. In the analyses presented here, we looked for the effects of smoking habits, alcohol consumption, vitamin uptake, chronic diseases, allergies, doing sports, gamma-GT (gamma-glutamyltranspeptidase), lymphocyte numbers, CEA (carcinoembryonic antigen), X-ray diagnostics, computer tomographies, and scintigraphies. With the exception of more than one scintigraphy carried out during the last four months before micronucleus analysis, none of the factors mentioned above significantly affected micronucleus numbers. One result deserves specific attention: individuals with low percentages of binucleated lymphocytes after in vitro cytochalasin B exposure showed higher micronucleus frequencies than those individuals with high percentages of binucleated cells. The same result was obtained for various other populations that we monitored in the past.

Topics: Alcohol Drinking; Alpha Particles; Cell Nucleus; Centromere; Chromosomal Instability; Cytochalasin B; Dose-Response Relationship, Radiation; Gamma Rays; Germany; Life Style; Lung Neoplasms; Lymphocytes; Micronucleus Tests; Mining; Occupational Diseases; Occupational Exposure; Pulmonary Fibrosis; Radionuclide Imaging; Radon; Risk Factors; Smoking; Time Factors; Uranium

We have previously established a vincristine resistant human lung cancer cell line (PC-9/VCR) by a stepwise exposure of parental line PC-9 to vincristine. In this study the resistant cells showed enhanced vincristine cytotoxicity in the presence of cytochalasin B and D. The increase in cytotoxicity was associated with an enhanced accumulation and a reduced efflux of vincristine. Colchicine and taxol had no effects on vincristine accumulation. Several cytoplasmic proteins were overexpressed in the resistant cells. The two major ones, with molecular weights of 58.8 kDa and 83.2 kDa, were shown by western blotting to be beta-tubulin and actin, respectively. The polymerized tubulin level in the resistant cells was significantly (p < 0.05) higher than that in the parental cells. These results suggest that the cellular cytoskeletons might play an important role in VCR resistance in the PC-9/VCR human lung cancer cell line.

Topics: Adenocarcinoma; Cell Survival; Colchicine; Cyclosporine; Cytochalasin B; Cytochalasin D; Cytoskeleton; Drug Resistance, Neoplasm; Drug Synergism; Humans; Lung Neoplasms; Neoplasm Proteins; Paclitaxel; Tubulin; Tumor Cells, Cultured; Verapamil; Vincristine

Human lung adenocarcinoma cells develop bipolar shape with prominent pseudopodia (greater than or equal to 200 microns) when cultured in the presence of autocrine motility factor (AMF)-like substance or on fibronectin-coated substrata. AMF was partially purified from a human lung adenocarcinoma cell line and has a peak biological activity at a molecular mass of 67 kDa. Using time-lapse photography, we observed that during AMF- or fibronectin-induced cell translocation, the nuclei of some bipolar cells are transported to the opposite end of the cell, while gross cell shape and position remain unchanged. Following this nuclear movement, which we call "nucleokinesis," the posterior pseudopodium is retracted behind the nucleus. Thus, extension of a pseudopodium followed by nucleokinesis in the same direction and retraction of the cell body behind the nucleus is a normal motile sequence in translocating bipolar cells. This suggests that nucleokinesis is a distinct step in whole-cell translocation of bipolar cells on biological substrata and that pseudopodia can be used as nuclear transport organs. In contrast, adenocarcinoma cells cultured on artificial substrata and in the absence of AMF display a fibroblast-like motility pattern with the nucleus centrally located within the migrating cell.

Topics: Adenocarcinoma; Cell Movement; Cell Nucleus; Colchicine; Culture Media; Cytochalasin B; Fibronectins; Glucose-6-Phosphate Isomerase; Growth Substances; Humans; In Vitro Techniques; Lung Neoplasms; Neoplasm Proteins; Tumor Cells, Cultured; Video Recording

Cytochalasin B (CB), at 100 or at 10 mg/kg single dose s.c. in carboxymethyl-cellulose (2%)/Tween-20 (1%) 24 h after s.c. challenge of B6D2F1 mice with trocar implants of B16F10 tumor s.c., delayed the appearance of measurable tumor nodules by 157 and 93%, respectively, and extended host survival by 65 and 26%. Tumor growth was also delayed when CB treatment was given 1 day after the appearance of palpable tumor nodules. By in vivo bioassay, in vitro cloning, and dye exclusion measurements, solid tumor nodules treated in vivo with CB at either 100 or 10 mg/kg showed the same viability and tumorigenicity as did vehicle-treated nodules 4 and 6 days after drug treatment, at which time growth inhibition was still apparent. This indicates that growth inhibition by CB is not dependent on a gross cytotoxic effect. CD2F1 mice challenged s.c. with Madison 109 lung carcinoma cells and treated with CB s.c. at 100 or 150 mg/kg 24 h later showed a 66% delay in the median day of tumor nodule appearance. When administered under these conditions or at the time of nodule appearance. CB markedly inhibited the rate of tumor growth, prevented tumor invasion at day 23, extended life span by 23%, and significantly inhibited spontaneous lung metastases measured 28 days after tumor challenge. Maximum tolerated doses of CB administered i.p., s.c., or i.v. in suspension or in solution are defined. These results delineate the conditions under which CB can be tested for in vivo biological activities and establish that this microfilament-active natural product in a single-agent protocol inhibits local tumor growth and extends survival in B16F10 melanoma and Madison 109 lung carcinoma, and, in the latter model, inhibits invasion and spontaneous lung metastases by mechanisms that do not appear to depend on cytotoxicity. This work on formulation, tolerated doses in vivo, and localized peritumoral effects of CB now permits evaluation of systemic antitumor effects of cytochalasin B as a single agent. It also permits chemotherapy studies using CB as a potential amplifier of the activity of other antitumor agents in vivo.

Topics: Animals; Cell Survival; Chromatography, Thin Layer; Cytochalasin B; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred BALB C; Neoplasm Invasiveness; Neoplasm Metastasis; Tumor Cells, Cultured

The human alveolar macrophage product, enzyme-releasing peptide (ERP), has a molecular mass of 8,000 Da, and releases azurophilic and specific granule constituents from neutrophils. A murine monoclonal anti-ERP antibody (12E10H), previously used to show a lack of antigenic identity between ERP and C5a, interleukin 1, tumor necrosis factor, and gamma-interferon, showed no cross-reactivity with interleukin 8. 12E10H and a fluorescein-labeled second antibody were used to visualize ERP on the macrophage surface. ERP was removed from alveolar macrophages by a 3-min incubation with 5 X 10(-7) M bovine pancreatic trypsin at 37 degrees C. The washed trypsinized cells could readhere to plastic and exclude trypan blue. Dilution of the trypsin-derived ERP released myeloperoxidase from cytochalasin-B-treated neutrophils dose dependently. The enzyme-releasing ability of the trypsin-derived material was removed by immunoprecipitation using antibody 12E10H bound to Staphylococcal protein A Sepharose 4B. The estimated molecular mass of the trypsin-derived ERP (by molecular sieve chromatography on HPLC) was approximately 8,500 Da. Other proteases (plasmin, thrombin, and cathepsin G) also released ERP from the cell surface, but the ERP was not an active secretagogue for neutrophils. However, macrophages cultured with protease inhibitors did not show decreased ERP accumulation in the medium. Our data indicate that ERP exists on the surface of human alveolar macrophages and can be released by proteases found within the lung environment in some disease states.

Topics: Antibodies, Monoclonal; Cell Membrane; Cells, Cultured; Chromatography, High Pressure Liquid; Cross Reactions; Cytochalasin B; Humans; Immunoblotting; Lung Neoplasms; Macrophages; Molecular Weight; Neutrophils; Peptides; Trypsin

Cloned metastatic Lewis lung carcinoma cells, C3, were more resistant to natural killer (NK) lysis than were nonmetastatic variant cells, C8. This was influenced by the tumor cell adhesiveness and morphology. When the nonadherent round C3 cells were cultured with dimethylsulfoxide, they became adherent and spread, sensitive to NK lysis and less metastatic. When the adherent and spread C8 cells were made nonadherent and round with cytochalasin B, they became more resistant to NK lysis and more metastatic. These metastatic differences were not observed in 3-week-old NK-deficient mice.

Topics: Animals; Cell Adhesion; Clone Cells; Cytochalasin B; Cytotoxicity, Immunologic; Dimethyl Sulfoxide; Female; Killer Cells, Natural; Lung Neoplasms; Mice; Mice, Inbred C57BL; Neoplasm Metastasis

A monoclonal antibody was developed against an 8,000-kDa enzyme-releasing peptide (ERP) released from human alveolar macrophages. ERP was isolated on an immunoaffinity column containing the antibody bound to staphylococcal protein A-Sepharose. Release of ERP from the macrophages is not changed by plastic adherence, phagocytosis, calcium ionophore, or phorbol esters. The peptide was not antigenically similar to interferon-gamma, tumor necrosis factor, or interleukin 1 alpha or 1 beta. The release of constituents from azurophilic and specific granules was the main identified biologic function of ERP. ERP was a more effective secretagogue in the untreated neutrophils and f-met-leu-phe was more effective in the cytochalasin B-treated neutrophils. Absorption of ERP from macrophage-conditioned medium removed a small amount of the chemotactic activity; however, the immunopurified peptide was not chemotactic or chemokinetic for neutrophils, and at high concentrations, it suppressed base line chemokinesis. Treatment of washed macrophages with trypsin released active ERP of approximately the same m.w. of spontaneously secreted ERP. These studies showed that human alveolar macrophages release a peptide which is a secretagogue for human neutrophils under conditions which may be encountered in the lungs during certain disease states. Proteolytic enzymes which are free in the lungs may release the peptide and lead to the secretion of neutrophil enzymes.

Topics: Antibodies, Monoclonal; Chemotaxis; Cytochalasin B; Cytoplasmic Granules; Exocytosis; Humans; Lung Neoplasms; Macrophages; Middle Aged; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Peptides; Pulmonary Alveoli; Trypsin

Topics: Carcinoma, Squamous Cell; Cell Cycle; Cell Line; Cytochalasin B; Diploidy; DNA Replication; Fetus; Humans; Lung; Lung Neoplasms; Nasopharyngeal Neoplasms

Liver tissue of a suckling rat was cultured. After 3 weeks of cultivation, the cultures consisting of epithelial cells were treated with 50 micrograms/ml or 100 micrograms/ml DEN for 7 days. 5 months after the treatment, the mode of chromosome number was found decreased from 42 to 40 in the 100 micrograms/ml DEN-treated group and shifted to triploid range after 21 months. The mode in the 50 micrograms/ml DEN-treated group maintained the diploid number until the 5th month but was found reduced to 40 in 21 months. On subcutaneous backtransplantation into young rats at the 22nd month, the treated cells produced tumors at the site inoculated in all the rats. Metastatic foci were also detected in lungs. These tumors were histologically diagnosed as hepatomas. Untreated control cells did not produce tumors. The differential effects of cytochalasine B on the cells with and without tumorigenicity were examined by the use of these cells and other cells, and it revealed that the capacity of multinucleated cell-formation by cytochalasin B fairly corresponds with the backtransplantability of the cells. Binucleated cell formation, not more than 2 nuclei, in the culture of normal cells was found by time-lapse cinemicrography to be not due to the non-capacity of multinucleation but to the destruction of multinucleated cells.

Topics: Animals; Animals, Suckling; Cell Nucleus; Cell Transformation, Neoplastic; Culture Techniques; Cytochalasin B; Diethylnitrosamine; Female; Liver; Liver Neoplasms, Experimental; Lung Neoplasms; Nitrosamines; Polyploidy; Rats

Topics: Adenocarcinoma; Cell Membrane; Cytochalasin B; Humans; Lung Neoplasms; Mesothelioma

Topics: Carcinoma; Cell Line; Cell Survival; Cells, Cultured; Cytochalasin B; Lung Neoplasms; Neoplasms, Experimental