Behavioural Differences And Cytogenetic Analysis Of A Transformed Cellular Population Derived From A Vero Cell Line.

Arising from a Vero cell line, an altered population was spontaneously obtained, which grew in multiple layers. Cytogenetic analyses revealed a higher polyploidy index (19%) than occurred in the initial population (5%). A modal chromosome number of 56 was found for the altered population, while the initial cell culture had only 54 chromosomes. Karyotypic studies showed that the extranumerary chromosomes were homologues of the 8th and the 27th pairs. The altered population probably resulted from a transformation of the initial line, and their transformed phenotypes may have been associated with the cytological differences.81324172

Cell Death Induced By Tamoxifen In Human Blood Lymphocytes Cultivated In Vitro [morte Celular Induzida Pelo Tamoxifeno Em Linfócitos Humanos Cultivados In Vitro]

Many chemotherapeutic agents with a potential against solid tumors or leukemia can cause lymphopenia. Tamoxifen (TAM) is a synthetic non-steroidal anti-estrogen drug employed in female breast cancer treatment. The present study investigated the capacity of TAM to induce cell death in human lymphocytes cultivated in vitro. Lymphocytes were obtained from young (25-30 years; n = 3) and elderly women (58-77 years; n = 3) and cultivated for 24 or 48h, with or without TAM (20 μM). After the culture, cell viability, immunocytochemical response and ultrastructure were evaluated. TAM affected lymphocytes in a time- dependent manner, and cells obtained from elderly women were the most sensitive to TAM. Immunocytochemical analysis evidenced higher frequency of apoptosis in treated cells, and the ultrastructural study revealed autophagic vacuoles, differing from the controls. In summary, the treated lymphocytes were affected by TAM, leading to cell death by apoptosis and autophagy.324415421Albukhari, A.A., Gashlana, H.M., Elbeshbishyb, H.A., Nagyc, A.A., Abdel-Naim, A.B., Caffeic acid phenethyl ester protects against tamoxifen-induced hepatotoxicity in rats (2009) Food and Chemical Toxicology, 47 (7), pp. 1689-1695Amarante-Mendes, G.P., Green, D.R., The regulation of apoptotic cell death (1999) Brazilian Journal of Medical and Biological Research, 32 (9), pp. 1053-1061Aycart, J.B., Pérez, I.S., Martín, T.R., Villaescusa, G., Andrade, M.C.G., Sarcomas de útero después de tratamiento con tamoxifeno por cancer de mama (2005) Oncología, 28 (7), pp. 38-42Baget-Bernaldiz, M., Soler, L.N., Romero-Aroca, P., Traveset-Maeso, A., Maculopatía por tamoxifeno: Estudio mediante la tomografía de coherencia óptica/ Optical coherence tomography study in tamoxifen maculopathy (2008) Archivos De La Sociedad Española De Oftalmologia, 83 (10), pp. 615-617Balducci, L., Corcoran, M., Antineoplastic chemotherapy of the older patient (2000) Hematology/oncology Clinics of North America, 14 (1), pp. 193-212Baral, E., Nagy, E., Kwok, S., McNicol, A., Gerrard, J., Berczi, I., Supression of lymphocytes mitogenesis by tamoxifen: Studies on protein kinase C, calmodulin and calcium (2000) Neuroimmunomodulation, 7 (2), pp. 68-76Carthew, P., Lee, P.N., Edwards, R.E., Heydon, R.T., Nolan, B.M., Martin, E.A., Cumulative exposure to tamoxifen: DNA adducts and liver cancer in the rat (2001) Archives of Toxicology, 75 (6), pp. 375-380Clemons, M., Danson, S., Howell, A., Tamoxifen (Nolvadex): A review (2002) Cancer Treatment Reviews, 28 (4), pp. 165-180Cui, Q., Tashiro, S., Onodera, S., Minami, M., Ikejima, T., Autophagy preceded apoptosis in oridonintreated human breast cancer MCF-7 cells (2007) Biological and Pharmaceutical Bulletin, 30 (5), pp. 859-864Daido, S., Kanzawa, T., Yamamoto, A., Takeuchi, H., Kondo, Y., Kondo, S., Pivotal role of the cell death factor BNIP3 in ceramide-induced autophagic cell death in malignant glioma cells (2004) Cancer Research, 64 (12), pp. 4286-4293Davis, W., Venitt, S., Phillips, D.H., The metabolic activation of tamoxifen and a-hydroxytamoxifen to DNAbinding species in rat hepatocytes proceeds via sulphation (1998) Carcinogenesis, 19 (5), pp. 861-866Drucker, L., Stackievicz, R., Radnay, J., Shapira, H., Cohen, I., Yarkoni, S., Tamoxifen enhances apoptotic effect of cisplatin on primary endometrial cell cultures (2003) Anticancer Research, 23 (2), pp. 1549-1554Friesen, C., Herr, I., Krammer, P.H., Debatin, K.M., Involvement of the CD95 (APO-1/FAZ) receptor/ligand system in drug-induced apoptosis in leukemia cells (1996) Nature Medicine, 2 (5), pp. 574-577Gil-Salú, J.L., Bosco-López, J., Domínguezvillar, M., Domínguez-Pascual, I., Pérezrequena, J., Palomo, M.J., López-Escobar, M., Ensayos de quimiosensibilidad en cultivos primarios de tumores cerebrales (2008) Neurocirugía, 19 (1), pp. 5-10Gozuacik, D., Kimchi, A., Autophagy as a cell death and tumor suppressor mechanism (2004) Oncogene, 12 (23), pp. 2891-2906Grubeck-Loebenstein, B., Berger, P., Saurwein-Teissl, M., Zisterer, K., Wick, G., No immunity for the elderly (1998) Nature Medicine, 4 (8), p. 870Hard, G.C., Iantropoulos, M.J., Jorda, K., Katenberger, O.P., Imondi, A.R., Williams, G.M., Major differences in the hepatocarcinogenecity and DNA adduct forming ability between toremofene and tamoxifen in female Crl (1993) CD(BR) Rats. Cancer Research, 53 (19), pp. 4534-4541Hemminki, K., Widlack, P., Hou, S.M., DNA adducts caused by tamoxifen and toremifene in human microssomal system and lymphocytes in vitro (1995) Carcinogenesis, 16 (7), pp. 1661-1664Hemminki, K., Rajaniemi, H., Koskinen, M., Hansoon, J., Tamoxifen-induced DNA adducts in leucocytes of breast cancer patients (1997) Carcinogenesis, 18 (1), pp. 9-13Jan, C.R., Cheng, J.S., Chou, K.J., Wang, S.P., Lee, K.C., Tang, K.Y., Tseng, L.L., Chiang, H.T., Dual effect of tamoxifen, an anti-breast-cancer drug, on intracellular Ca2+ and cytotoxicity in intact cells (2000) Toxicology and Applied Pharmacology, 168 (1), pp. 58-63Kim, H.S., Ishizaka, M., Kazusaka, A., Fujita, S., Di-(2-ethylhexyl) phthalate suppresses tamoxifeninduced apoptosis in GH3 pituitary cells (2007) Archives of Toxicology, 81 (1), pp. 27-33Klionsky, D.J., Cregg, J.M., Dunn, W.A., Emr, J.R.S.D., Sakai, Y., Sandoval, I.V., Sibirny, A., Ohsumi, Y., A unified nomenclature for yeast autophagy-related genes (2003) Developmental Cell, 5 (4), pp. 539-545Kondo, Y., Kondo, S., Autophagy and cancer therapy (2006) Autophagy, 2 (2), pp. 85-90Krysko, O., De, R.L., Cornelissen, M., Phosphatidylserine exposure during early primary necrosis (oncosis) in JB6 cells as evidenced by immunogold labeling technique (2004) Apoptosis, 9 (4), pp. 495-500Lamparska-Przybysz, M., Gajkowska, B., Motyl, T., Cathepsins and BID are involved in the molecular switch between apoptosis and autophagy in breast cancer MCF-7 cells exposed to camptothecin (2005) Journal of Physiology and Pharmacology, 56 (3), pp. 159-179Lecoeur, H., Prevost, M.C., Gougeon, M.L., Oncosis is associated with exposure of phosphatidylserine residues on the outside layer of the plasma membrane: A Reconsideration of the Specificity of the Annexin V/propidium Iodide Assay (2001) Cytometry, 44 (1), pp. 65-72Lichtman, S.M., Vilani, G., Chemotherapy in the elderly: Pharmacological considerations (2000) Cancer Control, 7 (6), pp. 548-556Majumdar, S.K., Valdellon, J.A., Brown, K.A., In vitro investigation on the toxicity and cell death induced by tamoxifen on two non-breast cancer cell types (2001) Journal of Biomedicine & Biotechnology, 1 (3), pp. 99-107Mandlekar, S., Kong, A.N., Mechanism of tamoxifen-induced apoptosis (2001) Apoptosis, 6 (6), pp. 469-477Matos, A.P., Paperna, I., Lainson, R., An erythrocytic virus of the brazilian tree-frog, Phrynohyas venulosa (1995) Memórias Do Instituto Oswaldo Cruz, 90 (5), pp. 653-655Matyja, E., Taraszewska, A., Naganska, E., Rafalowska, J., Autophagic degeneration of motor neurons in a model of slow glutamate excitotoxicity in vitro (2005) Ultrastructural Pathology, 29 (5), pp. 331-339Mizutani, A., Okada, T., Shibutani, S., Sonoda, E., Helfrid, H., Nishigori, C., Miyachi, Y.B., Yamazoe, M.A., Extensive Chromosomal Breaks Are Induced by Tamoxifen and Estrogen in DNA Repair-Deficient Cells (2004) Cancer Research, 64 (9), pp. 3144-3147Moretti, L., Yang, E.S., Kim, K.W., Lu, B., Autophagy signaling in cancer and its potential as novel target to improve anticancer therapy (2007) Drug Resistance Updates, 10 (4-5), pp. 135-143Nystedt, M., Berglund, G., Bolund, C., Fornander, T., Rutqvist, L.E., Side effects of adjuvant endocrine treatment in premenopausal breast cancer patients: A prospective randomized study (2003) American Journal of Clinical Oncology, 21 (9). , 1863-1844Ogata, A., Yanagie, H., Ishikawa, E., Morishita, Y., Mitsui, S., Yamashita, A., Hasumi, K., Eriguchi, M., Antitumour effect of polyoxomolybdates: Induction of apoptotic cell death and autophagy in in vitro and in vivo models (2008) British Journal of Cancer, 98 (2), pp. 399-409Omoti, A.E., Omoti, C.E., Toxicidad ocular de la quimioterapia sistémica anticancerosa (2006) Pharmacy Practice, 4 (2), pp. 55-59Osborne, C.K., Zhao, H., Fuqua, S.A., Selective estrogen receptor modulators: Structure, function, and clinical use (2000) Journal of Clinical Oncology, 18 (17), pp. 3172-3186Pagliara, P., Chionna, A., Panzarini, E., De, L.A., Caforio, S., Serra, G., Abbro, L., Dini, L., Lymphocytes apoptosis: Young versus aged and humans versus rats (2003) Tissue and Cell, 35 (1), pp. 29-36Petinari, L., Kohn, L.K., Carvalho, J.E., Genari, S.C., Cytotoxicity of tamoxifen in normal and tumoral cell lines and its ability to induce cellular transformation in vitro (2004) Cell Biology International, 28 (7), pp. 531-539Qadir, M.A., Kwok, B., Dragowska, W.H., To, K.H., Le, D., Bally, M.B., Gorski, S.M., Macroautophagy inhibition sensitizes tamoxifen-resistant breast cancer cells and enhances mitochondrial depolarization (2008) Breast Cancer Research and Treatment, 112 (3), pp. 389-403Sargent, L.M., Dragan, Y.P., Sattler, C., Bahnub, N., Sattler, G., Martin, P., Cisneros, A., Pilot, H.C., Induction of hepatic aneuploidy in vivo by tamoxifen, toremifene and idoxifene in female Sprague-Dawley rats (1996) Carcinogenesis, 17 (5), pp. 1051-1056Sawhney, R., Sehl, M., Naeim, A., Physiologic aspects of aging: Impact on cancer management and decision making, part I (2005) Cancer Journal, 11 (6), pp. 449-460Shahi, P.K., Izarzugaza, P.Y., Encinas, G.S., de la Díaz Muñoz, E.V.M., Pérez, M.G., Tratamiento adyuvante en el cancer de mama operable (2008) Anales De Medicina Interna, 25 (1), pp. 36-40Stahnke, K., Fulda, S., Friesen, C., Straub, G., Debatin, K.M., Activation of apoptosis pathways in peripheral blood lymphocytes by in vivo chemotherapy (2001) Blood, 98 (10), pp. 3066-3073Styles, J.A., Davies, A., Davies, R., White, I.N.H., Smith, L.L., Clastogenic and aneugenic effects of tamoxifen and some of its analogues in hepatocytes from doses rats and in human lymphoblastoid cells transfected with human P450 cDNAs (MCL-5 cells) (1997) Carcinogenesis, 18 (2), pp. 303-313Styles, J.A., Davies, R., Fenwick, S., Walker, J., White, I.N.H., Smith, L.L., Tamoxifen mutagenesis and carcinogenesis in livers of lambda/lacI transgenic rats: Selective influence of phenobarbital promotion 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Comparison of activation and detoxication mechanisms in rodents and in humans (2003) Current Drug Metabolism, 4 (3), pp. 223-239Wozniak, K., Kolacinska, A., Blasinskamorawiec, M., Morawiec-Bajda, A., Morawiec, Z., Zadrozny, M., Blasiak, J., The DNA-damaging potential of tamoxifen in breast cancer and normal cells (2007) Archives of Toxicology, 81 (7), pp. 519-52

Scanning And Transmission Electron Microscopy Of Transformed Vero Cells, With Altered In Vitro Growth Characteristics.

The Vero lineage, established from kidney cells of the green. African monkey, presented fibroblasts-like cells and growth in monolayers. Maintained in culture, the Vero cells presented behavioural and morphologic alterations, associated with cellular transformation. The morphological alterations were investigated using scanning and transmission electron microscopy. The study of proliferation and determination of the cellular doubling time was obtained from the growth curve. The initial population presented growth in a monolayer, while the altered cells grew in multilayers forming cellular aggregates, with flattened cells on the surface and globular cells in the inner region of the aggregate, together with extracellular matrix material. The cell surface of the altered population presented innumerable structures similar to little vesicles, microvilli and cytoplasmic prolongations. The cellular proliferation of both populations was very similar. Our results indicate that morphological and growth changes probably resulted from cellular transformation of the initial Vero cells. These transformed cells presented several characteristics associated with neoplastic growth, and can be used as a model for tumor cells studies in vitro.28456557

Assessment Of The Possible Toxic Effect Of A Semifluorinated Alkane On Vero Cell Culture [avaliação Do Possível Efeito Tóxico De Um Alcano Semifluorinado De Uso Oftalmológico Sobre Cultura De Células Vero]

Purpose: Perfluorocarbon liquids (PFCLs) are used in vitreoretinal surgery. PFCLs may cause inflammatory reactions, cellular injury and destruction of the normal retinal architecture. In order to avoid these effects, semifluorinated alkanes (SFA) were developed. We assessed the potential use of an SFA known as perfluorohexiloctano (F6H8) as long-term vitreous replacement under controlled cell culture conditions. Methods: We analyzed indirect cytotoxicity, where the cells only come into contact with soluble elements that can be eliminated by perfluorohexiloctano. We therefore analyzed direct toxicity (contact toxicity) of perfluorohexiloctano by means of scanning electronic microscopy and immunocytochemistry reagents for actin. Cells embedded in a treatmentfree culture medium were used as control, a positive control for toxicity with an undeniably toxic effect on cells, and a weight control that produced a mechanical compression similar to the amount of perfluorohexiloctano used in the experiment. Results: The indirect cytotoxicity test showed that F6H8 did not affect cell growth. Our direct toxicity tests showed that cellular alterations caused by perfluorohexiloctano were similar to those produced by the weight control and different from toxicity control. Conclusions: Perfluorohexiloctano does not present indirect toxicity and this product has a compressive rather than a toxic effect on cultured cells.676905910Mertens, S., Bednarz, J., Richard, G., Engelmann, K., Effect of perfluorodecalin on human retinal pigment epithelium and human corneal endothelium in vitro (2000) Graefes Arch Cli Exp Ophthalmol., 238 (2), pp. 181-185Mathis, A., Pagot, V., Gazagne, C., Malecaze, F., Giant retinal tears. Surgical techniques and results using perfluorodecalin and silicone oil tamponade (1992) Retina, 12 (SUPPL. 3), pp. S7-10Chang, S., Reppucci, V., Zimmerman, N.J., Heinemann, M.H., Coleman, D.J., Perfluorocarbon liquids in the management of traumatic retinal detachments (1989) Ophthalmology, 96 (6), pp. 785-792Glaser, B.M., Carter, J.B., Kuppermann, B.D., Michels, R.G., Perfluoro-octane in the treatment of giant retinal tears with proliferative vitreoretinopathy (1991) Ophthalmology, 98 (11), pp. 1613-1621Chang, S., Ozmert, E., Zimmerman, N.J., Intraoperative perfluorocarbon liquids in the management of proliferative vitreoretinopathy (1988) Am J Ophthalmol, 106 (6), pp. 668-674Moreira Junior, C.A., Uscocovich, C.E., Moreira, A.T., Experimental studies with perfluoro-octane for hemostasis during vitreoretinal surgery (1997) Retina, 17 (6), pp. 530-534Sparrow, J.R., Jayakumar, A., Berrocal, M., Ozmert, E., Chang, S., Experimental studies of the combined use of vitreous substitutes of high and low specific gravity (1992) Retina, 12 (2), pp. 134-140Liang, C., Peyman, G.A., Tolerance of extended-term vitreous replacement with perfluoro-n-octane and perfluoroperhydrophenanthrene mixture (phenoctane) (1999) Retina, 19 (3), pp. 230-237Loewenstein, A., Humayun, M.S., De Juan Jr., E., Campochiaro, P.A., Haller, J.A., Perfluoroperhydrophenanthrene versus perfluoro-n-octane in vitreoretinal surgery (2000) Ophthalmology, 107 (6), pp. 1078-1082Estacia, P., Rodrigues Jr., A.S., Moreira, P.L., Genari, S.C., The cytotoxicity in vero cells of a perfluorocarbon used in vitreoretinal surgery (2000) Braz J Morphol Sci., 19 (2), pp. 41-47Chang, S., Sparrow, J.R., Iwamoto, T., Gershbein, A., Ross, R., Ortiz, R., Experimental studies of tolerance to intravitreal perfluoro-n-octane liquid (1991) Retina, 11 (4), pp. 367-374Meller, D., Augustin, A.J., Spitznas, M., Lutz, J., Meller, K., Effect of different perfluorochemicals on dorsal root ganglion cells in vitro (1998) Graefes Arch Clin Exp Ophthalmol, 236 (3), pp. 182-187Winter, M., Winter, C., Wiechens, B., Quantification of intraocular retained perfluorodecalin after macroscopic complete removal (1999) Graefes Arch Clin Exp Ophthalmol, 237 (2), pp. 153-156Velikay, M., Wedrich, A., Stolba, U., Datlinger, P., Li, Y., Binder, S., Experimental long-term vitreous replacement with purified and nonpurified perfluorodecalin (1993) Am J Ophthalmol, 116 (5), pp. 565-570Kirchhof, B., Fluorocarbones in vitreoretinal surgery (1999) Ophtalmo-chirurgie, 11, pp. 153-158Singer, H., New studies further define role of PFCLs in vitreoretinal surgery (1999) Ocular Surg News, 17, p. 50. , FebLangefeld, S., Kirchhof, B., Meinert, H., Roy, T., Aretz, A., Schrage, N.F., A new way of removing silicone oil from the surface of silicone intraocular lenses (1999) Graefes Arch Clin Exp Ophthalmol, 237 (3), pp. 201-206Wong, D., Lois, N., Perfluorocarbons and semifluorinated alkanes (2000) Semin Ophthalmol, 15 (1), pp. 25-35. , ReviewZeana, D., Becker, J., Kuckelkorn, R., Kirchhof, B., Perfluorohexiloctano as long-term vitreous tamponade in the experimental animal. Experimental perfluorohexiloctano substitution (1999) Int Ophthalmol, 23 (1), pp. 17-24Kirchhof, B., Wong, D., Van Meurs, J., Hilgers, R.D., Macek, M., Lois, N., Schrage, N.F., Use of perfluorohexiloctano as a long-term internal tamponade agent in complicated retinal detachment surgery (2002) Am J Ophthalmol, 133 (1), pp. 95-101Stefaniotou, M.I., Aspiotis, M.V., Kitsos, G.D., Kalogeropoulos, C.D., Asproudis, I.C., Psilas, K.G., Our experience with perfluorohexiloctano (F6H8) as a temporary endotamponade in vitreoretinal surgery (2002) Eur J Ophthalmol, 12 (6), pp. 518-522Vote, B., Wheen, L., Cluroe, A., Teoh, H., McGeorge, A., Further evidence for proinflammatory nature of perfluorohexiloctano in the eye (2003) Clin Experiment Ophthalmol, 31 (5), pp. 408-414(1992) Biological evaluation of medical devices - Part. 5. - Tests for cytotoxicity: "in vitro" methods, , ISO 10993-1Murakami, N., Fukuchi, S., Takeuchi, K., Hori, T., Shibamoto, S., Ito, F., Antagonistic regulation of cell migration by epidermal growth factor and glucocorticoid in human gastric carcinoma cells (1998) J Cell Physiol, 176 (1), pp. 127-137Peyman, G.A., Schulman, J.A., Sullivan, B., Perfluorocarbon liquids in ophthalmology (1995) Surv Ophthalmol, 39 (5), pp. 375-395. , ReviewChang, S., Low viscosity liquid fluorochemicals in vitreous surgery (1987) Am J Ophthalmol, 103 (1), pp. 38-43Eckardt, C., Nicolai, U., Winter, M., Knop, E., Experimental intraocular tolerance to liquid perfluorooctane and perfluoropolyether (1991) Retina, 11 (4), pp. 375-384Stolba, U., Krepler, K., Pflug, R., Velikay, M., Wedrich, A., Binder, S., Experimental vitreous and aqueous replacement with perfluorophenanthrene (1997) Retina, 17 (2), pp. 146-153Green, K., Slagle, T., Chaknis, M.J., Cheeks, L., Chang, S., Perfluorocarbon effects on rabbit blood-retinal barrier permeability (1993) Ophthalmic Res., 25 (3), pp. 186-191Meinert, H., Roy, T., Semifluorinated alkanes - A new class of compounds with outstanding properties for use in ophthalmology (2000) Eur J Ophthalmol, 10 (3), pp. 189-19

Cisplatin-induced Cytogenetic Alterations In V79 Cells

Cisplatin is an antineoplastic agent used to treat solid malignancies, such as ovarian, testicular and bladder tumors. Studies have been shown that cisplatin induces genotoxic effects and chromosomal alterations that can result in genetic and chromosomal instability. In this work, we investigated the effects of cisplatin on the cytogenetic traits of cultured V79 cells based on the modal chromosome number, mitotic index, frequency of polyploidy and number of aneuploid metaphases. Cisplatin-treated V79 cells showed an altered chromosome number distribution, as well as an enhanced mitotic index, frequency of polyploidy and number of aneuploid metaphases. These alterations were probably related to the genetic instability produced by cisplatin. In addition, these cells showed characteristics associated with neoplastic development that corresponded to neoplastic processes related to the use of cisplatin in chemotherapy. © 2007 The Japan Mendel Society.722155160Allavena, P., Pirovano, P., Bonazzi, C., Colombo, N., Mantovani, A., D'Incalci, M., In vitro and in vivo effects of cisplatin on the generation of lymphokine-activated killer cells (1990) J. Natl. Cancer Inst, 82, pp. 139-142Barrett, J.C., Cell culture models of multistep carcinogenesis (1985) IARC Sci. Publ, 58, pp. 181-202Borenfreund, E., Babich, H., Martin-Alguacil, N., Effect of methylazoxymethanol acetate on bluegill sunfish cell cultures in vitro (1989) Ecotoxicol. Environ. Saf, 17, pp. 297-307Bradley, M.O., Bhuyan, B., Francis, M.C., Langenbach, R., Peterson, A., Huberman, E., Mutagenesis by chemical agents in V79 chinese hamster cells: A review and analysis of the literature. A report of the Gene-Tox Program (1981) Mutat. 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