RESUMEN
PURPOSE: Idazoxan hydrochloride (IDA) is a 241 molecular weight imidazoline and adrenoreceptor ligand. It binds to mitochondrial membranes and promotes apoptosis of pancreatic beta cells. Since IDA has not been tested against tumor cells, the purpose of our study was to determine if IDA has antineoplastic activity. METHODS: We used the conversion of a soluble tetrazolium salt to an insoluble formazan precipitate and differential staining cytotoxicity assays to determine if IDA was cytotoxic to cell lines of murine lung cancer and human prostate cancer, as well as to a variety of fresh human tumor samples. We used flow cytometry to analyze cell death and calreticulin expression. RESULTS: IDA is cytotoxic to both cell lines and against aliquots of specimens of breast, gastric, lung, ovarian and prostate cancers as well as non-Hodgkin's lymphoma. It produces apoptotic cell death and promotes calreticulin expression, suggesting that IDA might be immunomodulatory in vivo. CONCLUSION: We anticipate that IDA will be clinically useful in cancer treatment.
Asunto(s)
Antineoplásicos/farmacología , Idazoxan/farmacología , Neoplasias/patología , Animales , Anexina A5/metabolismo , Apoptosis/efectos de los fármacos , Calreticulina/metabolismo , Carcinoma Pulmonar de Lewis/patología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Masculino , Ratones , Neoplasias de la Próstata/patología , Células Tumorales CultivadasRESUMEN
PURPOSE: To determine if the antineoplastic effect of etoposide includes alteration in Lewis lung cancer cells which evoke an immunologic response in C57B1/6 host mice. METHODS AND RESULTS: Of C57B1/6 mice injected with 10(6) Lewis lung cancer (3LL) cells followed by treatment with a single 50 mg/kg dose of etoposide (VP-16), 60% survived over 60 days, in contrast to untreated control mice which died within 30 days. Approximately 40% of surviving mice rejected a subsequent challenge with 3LL. Their splenocytes protected naive mice injected with 3LL. To test if VP-16 treatment produced alterations in 3LL cells, which induce host immunity, leading to tumor rejection, C57B1/6 mice were injected with 3LL cells that had survived an 80-90% lethal concentration of VP-16 in vitro. These cells killed 75% of recipient mice but 60% of the surviving mice rejected challenge with 3LL. Splenocytes harvested from tumor-rejecting mice protected naive mice injected with 3LL. CONCLUSION: These results support the hypothesis that in addition to its antineoplastic cytotoxic effect, VP-16 induces changes in 3LL cells which are recognized by the host immune system resulting in immune rejection of 3LL. often immunosuppressive and therapeutic advantage is generally based on the tumor cytotoxicity of individual drugs or combinations of drugs [13]. Our earlier work showed a link between the use of cytotoxic chemotherapy with etoposide (VP-16) and the induction of an immune response against syngeneic murine leukemia in the intact host [16]. VP-16 is an immunosuppressive topoisomerase II-inhibiting drug which induces tumor cell apoptosis and is frequently used clinically to treat a variety of tumors [1, 3, 9, 10]. We have noted that the addition of cyclosporin A to VP-16 produces CD8 T lymphocyte-mediated tumor-specific immunity in mice bearing L1210 leukemia [17]. We have extended these experiments to a spontaneously arising non-carcinogen-induced neoplasm, Lewis lung cancer (3LL), and now report that surviving mice successfully treated with VP-16, in the absence of cyclosporin A, reject challenge with 3LL. In addition, results are presented to show that VP-16 modifies 3LL cells rendering them immunogenic. These findings are submitted to support the hypothesis that VP-16-induced cytotoxic changes include cellular membrane alterations in 3LL cells which are recognized by the immune system and cause rejection of this syngeneic lung tumor.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Carcinoma Pulmonar de Lewis/tratamiento farmacológico , Carcinoma Pulmonar de Lewis/inmunología , Etopósido/farmacología , Inmunidad Celular/efectos de los fármacos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/inmunología , Animales , Carcinoma Pulmonar de Lewis/patología , Membrana Celular/inmunología , Ensayo de Inmunoadsorción Enzimática , Rechazo de Injerto , Humanos , Inmunoglobulina G/análisis , Neoplasias Pulmonares/patología , Ratones , Ratones Endogámicos C57BL , Bazo/citología , Trasplante HeterólogoAsunto(s)
Riñón/patología , Síndrome Nefrótico/patología , Enfermedad Aguda , Adolescente , Biopsia , Diagnóstico Diferencial , Femenino , Glomerulonefritis Membranosa/complicaciones , Glomerulonefritis Membranosa/patología , Glomeruloesclerosis Focal y Segmentaria/complicaciones , Glomeruloesclerosis Focal y Segmentaria/patología , Humanos , Síndrome Nefrótico/etiologíaRESUMEN
PURPOSE: The overall purpose of this study was to determine the potential efficacy of epoxide-containing piperazines as a new class of anti-cancer agents. Two representative compounds, specifically NCO-700, a 4-trimethoxyphenyl-substituted epoxide-piperazine, and TOP-008, a 4-phenylpropenyl-substituted epoxide-piperazine were tested in cytotoxic assays with human breast and prostate cancer cell lines. A second objective was to determine if these two compounds had anti-cancer activity in vivo when tested against xenograft tumors in nude mice or human tumors grown under the kidney capsule in mice. A final objective of this study was to establish if NCO-700 and TOP-008 achieved cancer cell killing through an apoptotic mechanism. METHODS: The anti-proliferative activity of NCO-700 and TOP-008 were tested in a 7 day cell-survival assay utilizing a number of well characterized breast (HS-578T, T47D, MCF-7) and prostate (DU-145, PC-3, LNCaP) cancer cell lines. In vivo studies with the two compounds were performed, in nude mice bearing DU-145 xenograft tumors, and in normal mice in which DU-145 prostate cancer cells and HS-578T breast cancer cells were grown as solid tumors in the subrenal capsules of the animals. Apoptotic cell death of cancer cells was determined by a number of established techniques that detect apoptosis, including the confocal laser microscopy of treated cells and mitochondrial leakage assays utilizing the cationic dye, JC-1. Finally, the activation of the caspase cascade, enzymes that carry out apoptosis in mammalian cells, was examined in treated cells by immunoblot assays. RESULTS: NCO-700 and TOP-008 displayed cytotoxicity to HS-578T human breast cancer cells, with ED(50) values in the 3-6 microM range. Cytotoxicity to androgen receptor-negative human prostate cancer cells (PC-3 and DU-145 cells) occurred with ED(50) values in the 5-20 microM range. Cytotoxicity to hormone receptor-positive breast and prostate cancer cell lines occurred at 10 to 20-fold higher concentrations of the two compounds. When human prostate (DU-145) or breast cancer (HS-578T) cells were grown as solid tumors in the subrenal capsules of mice, significant anti-tumor activity of NCO-700 was observed at 20 mg/kg and 50 mg/kg body weight respectively, for prostate and breast tumors. In nude mice bearing DU-145 prostate tumor xenografts, 50 mg/kg doses of the two compounds either stopped (TOP-008) tumor growth or slowed (NCO-700) growth. The mechanism of cytotoxicity was shown to be through apoptosis, (a) by confocal microscopy studies revealing nuclear fragmentation, (b) by mitochondrial studies revealing disruption of the mitochondrial membrane and release of the cationic dye, JC-1, into the cytoplasm and (c) by protein immunoblot assays indicating that over a 6 h period, TOP-008 induced a significant accumulation of the pro-apoptotic protein, bak, in the mitochondrial fraction of HS-578T human breast cancer cells, accompanied by activation, at 2.5 h, of caspase-3. CONCLUSIONS: These studies indicated that the epoxide-containing piperazines, as exemplified by NCO-700 and TOP-008, were effective anti-cancer agents when tested in vitro and in vivo against human breast and prostate tumors. Our studies also indicated that TOP-008 induced the initiation of the caspase cascade leading to apoptosis. Previous toxicology studies in rodents and dogs, as well as a Phase I study in humans, showed NCO-700 to be a well-tolerated, non-toxic compound. Taken together with our current findings, these results suggest that this class of compounds has the potential to be relatively safe, new chemotherapeutic agents for refractory breast and prostate cancers.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Piperazinas/farmacología , Inhibidores de Proteasas/farmacología , Animales , Bencimidazoles/metabolismo , Carbocianinas/metabolismo , Caspasa 3 , Caspasas/metabolismo , División Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Compuestos Epoxi/química , Citometría de Flujo , Humanos , Immunoblotting , Riñón/efectos de los fármacos , Riñón/patología , Proteínas de la Membrana/metabolismo , Ratones , Ratones Desnudos , Microscopía Confocal , Mitocondrias/metabolismo , Trasplante de Neoplasias , Neoplasias Experimentales/tratamiento farmacológico , Neoplasias Experimentales/patología , Óxidos/química , Óxidos/farmacología , Piperazinas/química , Trasplante Heterólogo , Células Tumorales Cultivadas/citología , Células Tumorales Cultivadas/efectos de los fármacos , Proteína Destructora del Antagonista Homólogo bcl-2RESUMEN
We and others have shown that cyclosporin A (CsA) reverses resistance to etoposide (E) and cis-platinum (P) in vitro and in vivo. To assess the clinical relevance of combined therapy, we studied CsA with EP in patients with advanced non-small cell lung cancer in a phase I/II clinical trial in a University setting. Patients were treated between July 1989 and June 1994 and included 10 females and 34 males with a median age of 61 years and a mean Karnofsky performance status of 80. CsA was given at escalating doses of 1-6 mg/kg per day on days 1-4 of each 21 day cycle with cis-platinum 25 mg/m2 per day and etoposide 100 mg/m2 per days on days 1-3. Response was assessed after each 2 cycles by measuring index lesions. A total of 44 patients received 133 cycles, 22.7% of patients had a partial response and 36.4% had stable disease with 8% 2-year survival. Patients receiving 1-2 mg/kg CsA had a PR rate of 37.5 and 50% SD compared to 19.4 and 33.3% for doses of 3 mg/kg or more. Although no conclusions should be drawn from this small study, the Kaplan-Meier survival curves were statistically significant different for these two groups by the log-rank test (P = 0.047). The 2-year survival of the former group was 25% compared to 4% for the latter. In light of the potential importance of immunomodulation in cancer control, it seems prudent to balance the effects of CsA on P-glycoprotein and other drug resistance pumps against its dose-dependent immunosuppressive activity. Further studies are needed to validate the activity of low dose CsA in combination with standard chemotherapy for lung cancer.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Ciclosporina/administración & dosificación , Etopósido/administración & dosificación , Neoplasias Pulmonares/tratamiento farmacológico , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carcinoma de Pulmón de Células no Pequeñas/mortalidad , Ciclosporina/efectos adversos , Etopósido/efectos adversos , Femenino , Humanos , Neoplasias Pulmonares/mortalidad , Masculino , Persona de Mediana Edad , Tasa de SupervivenciaRESUMEN
PSC-833, a non immunosuppressive analogue of cyclosporin A, is an effective modulator of the multidrug-resistant tumor phenotype. Since both PSC-833 and cyclosporin A also enhance the cytotoxicity of VP-16 against drug sensitive L1210 leukemia cells in vitro we compared these agents as modulators of VP-16 efficacy in vivo. Compared to VP-16 treatment alone both PSC-833 and cyclosporin A significantly altered the survival of L1210 leukemia-bearing BDF/1 mice and Lewis lung carcinoma-bearing C57/B1 mice. Cyclosporin A enhanced VP-16 efficacy whereas PSC-833 impaired VP-16 efficacy against these murine tumors. Possible reasons for these disparate effects are discussed.
Asunto(s)
Ciclosporina/uso terapéutico , Ciclosporinas/uso terapéutico , Etopósido/uso terapéutico , Inmunosupresores/uso terapéutico , Leucemia L1210/tratamiento farmacológico , Neoplasias Pulmonares/tratamiento farmacológico , Animales , Antineoplásicos Fitogénicos/uso terapéutico , Sinergismo Farmacológico , Femenino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Tasa de Supervivencia , Factores de TiempoRESUMEN
Multidrug resistance is probably the single greatest obstacle to successful systemic therapy of human cancer. We have reported that cyclosporin A (CsA) can overcome multidrug resistance and improve the efficacy of etoposide in a murine model of drug-sensitive leukemia. The combination of CsA and paclitaxel (PCL) was also significantly superior to either drug alone against murine P388 (sensitive) and L1210 (resistant) leukemia. Lung cancer cells provide an ideal model system to study this phenomenon because both de novo and acquired drug resistance occur. Standard chemotherapy for advanced lung cancer is poorly effective, and although PCL is one of the most active new agents for this disease, responses occur in only 20% of patients. In vitro, CsA significantly enhanced the efficacy of PCL against lung (Lu-CSF-1 and 3LL) and oropharyngeal (CSCC-20) cancer cell lines. The combination also produced an increase in expression of interleukin 1beta, a cytokine known to inhibit the growth of Lu-CSF-1 cells. CsA alone had little or no antiproliferative activity in vitro and did not alter PCL transport. These results indicate that the activity of chemotherapy modulators may extend beyond mitigation of drug resistance to enhancement of therapeutic efficacy against drug-sensitive tumor cells in vitro and in vivo.
Asunto(s)
Antineoplásicos Fitogénicos/uso terapéutico , Ciclosporina/uso terapéutico , Inmunosupresores/uso terapéutico , Leucemia/tratamiento farmacológico , Paclitaxel/uso terapéutico , Neoplasias del Sistema Respiratorio/tratamiento farmacológico , Animales , Citocinas/biosíntesis , Modelos Animales de Enfermedad , Resistencia a Múltiples Medicamentos , Resistencia a Antineoplásicos , Sinergismo Farmacológico , Glicerol/análogos & derivados , Glicerol/farmacología , Sustancias de Crecimiento/metabolismo , Humanos , Leucemia/inmunología , Leucemia/mortalidad , Ratones , Neoplasias del Sistema Respiratorio/inmunología , Tasa de Supervivencia , Resultado del Tratamiento , Células Tumorales CultivadasRESUMEN
The relationship between the nuclear morphology and the microtubular organization of L100 and L1000 cells, two vincristine-induced multidrug resistant human acute lymphocytic leukemia cell lines, was examined and compared to that of L0 parental cells. The L0 parental cells contained a round nucleus and the microtubules were evenly distributed throughout the cytoplasm. In contrast, the microtubules of the L100 and L1000 cells were localized between the lobular structures of a multilobulated nucleus. Disassembly of microtubules in L100 and L1000 cells by colchicine resulted in the loss of the multilobulated morphology of the nucleus. While the total cellular content of tubulin of L0 and L100 cells was similar, the content of microtubules of L100 cells was only 55% of that observed in L0 cells. Two, 28 kDa (pI 6.9) and 31 kDa (pI 4.4), microtubule-associated proteins were found to be overexpressed in L100 and L1000 cells. The results indicate that the multilobulated nuclear morphology of L100 and L1000 cells is dependent upon the unique and intact organization of the microtubules; the distinct organization of the microtubules and the multilobular nuclear morphology of the two resistant cell lines may be due to the differential expression of specific microtubule-associated proteins.
Asunto(s)
Núcleo Celular/patología , Resistencia a Múltiples Medicamentos , Microtúbulos/patología , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Ciclo Celular , Núcleo Celular/ultraestructura , Electroforesis en Gel Bidimensional , Humanos , Microscopía Confocal , Proteínas Asociadas a Microtúbulos/análisis , Microtúbulos/ultraestructura , Tubulina (Proteína)/análisis , Células Tumorales CultivadasRESUMEN
The role of the immune response in the chemotherapeutic cure of an intact host with neoplasia is not well defined. We have previously shown that the addition of cyclosporin A to VP-16 therapy of BDF/1 mice with L1210 leukemia produces immunity to leukemia in long-surviving host animals. We now characterize this immunity as being tumor specific and related to the participation of CD8 T-lymphocytes. Splenocytes derived from L1210 leukemia immune mice are cytotoxic to L1210 cells after in vitro restimulation, compared to splenocytes harvested from nonimmune control mice. This cytotoxicity is lost by CD8 T-lymphocyte depletion and persists in Ia antigen blocking experiments. Cytotoxicity is selective for L1210 leukemia compared to P388 leukemia, an alternate Ia antigen expressing methylcholanthrine-induced acute lymphoid leukemia, and L1210 leukemia-immune mice remain susceptible to P388 leukemia in vivo demonstrating specificity of the immune response generated by cyclosporin A/VP-16 therapy.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Ciclosporina/administración & dosificación , Etopósido/administración & dosificación , Leucemia L1210/inmunología , Linfocitos T Citotóxicos/inmunología , Animales , Citotoxicidad Inmunológica , Inmunidad Celular , Inmunización Pasiva , Leucemia L1210/tratamiento farmacológico , Depleción Linfocítica , Ratones , Bazo/inmunologíaRESUMEN
The purpose of this study was to determine if a radiolabeled murine monoclonal antibody (EOS) directed against eosinophil peroxidase would localize specifically to tumor sites in patients with lymphomas infiltrated by eosinophils. Ten patients with Hodgkin's disease and eosinophilia, three patients with non-Hodgkin's lymphomas and eosinophilia and five control patients received an intravenous injection of 3-10 mg of EOS antibody radiolabeled with 74-155 MBq (2.0-4.2 mCi) of 111In. At intervals of 24, 48 and 72 hr after injection, gamma camera images were obtained along with blood and urine specimens and the imaging results were correlated with the results of other staging modalities. As early as 24 hr after antibody injection, there was clear visualization of identifiable sites of lymphoma with eosinophilia greater than 1 cm in size, including the spleen, bone marrow and lymph nodes. Although EOS also localized nonspecifically to the liver and, in some patients, to the nasopharynx, there was no appreciable uptake in normal bone marrow, spleen, uninvolved lymph nodes, lymphomas without eosinophilia or various other pathologic conditions without eosinophilia. Except for transient pain at tumor sites in three patients, no adverse reactions were noted. We conclude that a radiolabeled monoclonal antibody directed against eosinophil peroxidase localizes to lymphoma sites infiltrated by eosinophils.
Asunto(s)
Enfermedad de Hodgkin/diagnóstico por imagen , Linfoma no Hodgkin/diagnóstico por imagen , Radioinmunodetección , Adulto , Anciano , Femenino , Humanos , Radioisótopos de Indio , Masculino , Persona de Mediana EdadRESUMEN
Acquired resistance to chemotherapeutic drugs by tumor cells is an important obstacle to effective therapy of human malignancy. We now describe a vincristine (VCR)-induced multidrug-resistant (MDR) human acute lymphatic leukemia cell line, the sustained in vitro growth of which is dependent on vincristine. The doubling time for parental drug-sensitive cells (L0) is 40.2 +/- 13.2 h and for the MDR subline (L100) 62.5 +/- 11.3 h. L100 cells have similar G2 and mitotic phase to parental cells, express the MDR phenotype, and are characterized by novel morphologic features with multilobulated nuclei and multiple small nucleoli. Compared with L0 cells which have 2-3 nucleoli per cell, L100 cells have 7-8 nucleoli per cell. Average nucleolar area is 11.3 +/- 7.3 microns 2 for L0 and 2.5 +/- 2.4 microns 2 for L100 cells determined by the laser scanning method. The striking morphologic abnormalities of L100 cells suggest a drug-induced cytoskeletal abnormality. The relationship of these abnormalities to the VCR growth dependence of L100 cells is discussed.
Asunto(s)
Nucléolo Celular/ultraestructura , Núcleo Celular/ultraestructura , Leucemia-Linfoma Linfoblástico de Células Precursoras/patología , Vincristina/farmacología , Ciclo Celular/efectos de los fármacos , Colchicina/farmacología , Citoesqueleto/efectos de los fármacos , Resistencia a Medicamentos , Técnica del Anticuerpo Fluorescente , Humanos , Fenotipo , Células Tumorales CultivadasRESUMEN
We studied the effects of cyclosporin A and verapamil on the modulation of vincristine and daunorubicin resistance in a multidrug-resistant subline of human T-cell acute lymphatic leukemia GM3639. Our results show that cyclosporin A is more effective than verapamil as a modulator of the high degree of primary vincristine resistance and the low degree of daunorubicin cross-resistance expressed by this cell line. Isobologram analysis revealed that the combined modulators act synergistically in correcting both vincristine and daunorubicin resistance.
Asunto(s)
Ciclosporina/farmacología , Daunorrubicina/farmacología , Leucemia-Linfoma de Células T del Adulto/tratamiento farmacológico , Verapamilo/farmacología , Vincristina/farmacología , Resistencia a Medicamentos , Sinergismo Farmacológico , Humanos , Cinética , Leucemia-Linfoma de Células T del Adulto/patología , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Our prior in vitro studies on the correction of multidrug resistance by cyclosporin A (CsA) prompted us to investigate the effect of CsA and VP-16 in vivo. CsA given simultaneously at 2 or 10 mg/kg with VP-16 to BDF/1 mice bearing parental drug-sensitive P388 or L1210 lymphatic leukemia produced a 100% increase in survival as compared with VP-16 treatment alone. CsA-containing regimens also promoted 60-day survival in a significant number of P388 or L1210 leukemia-bearing mice as compared with animals receiving VP-16 in the absence of CsA (P < 0.02 and P < 0.001, respectively). CsA enhancement of the survival of mice bearing these lymphatic leukemias is restricted to VP-16, since the addition of CsA to therapeutic agents such as vincristine, daunorubicin, methotrexate, or cisplatin had no effect on survival.
Asunto(s)
Ciclosporina/farmacología , Etopósido/farmacología , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Animales , Sinergismo Farmacológico , Femenino , Leucemia L1210/tratamiento farmacológico , Leucemia L1210/mortalidad , Leucemia P388/tratamiento farmacológico , Leucemia P388/mortalidad , Ratones , Ratones Endogámicos , Leucemia-Linfoma Linfoblástico de Células Precursoras/mortalidad , Tasa de Supervivencia , Factores de TiempoRESUMEN
Cyclosporin A (CsA) is an effective modulator of multidrug resistance (MDR) in vitro and in murine tumour systems in vivo. We now report the production of immunity to L1210 leukaemia by the addition of CsA to VP-16 therapy of leukaemic BDF/1 mice. VP-16/cyclosporin A tumour immunity induction arises as a consequence of active therapy independently of immunisation with modified tumour cells. The addition of CsA to VP-16 prolongs survival of BDF/1 host mice bearing L1210 leukaemia beyond that produced by equivalent dose VP-16 alone. A subpopulation of 60-day surviving mice after combined VP-16/CsA are immune to rechallenge with the same leukaemia inoculum to which they were originally exposed. Spleen cells from immune mice adoptively transfer anti-L1210 leukaemia immunity to untreated BDF/1 mice in a dose dependent, statistically significant manner. Adoptive transfer experiments additionally suggest active recruitment of immunologic response in recipient animals: (1) We have been able to perpetuate leukaemia immunity in four sequential cohorts of naive recipient mice. This propogation of adoptive immunity is accomplished by use of spleen cells harvested from each preceeding passively-protected animal cohort; (2) Cyclophosphamide pretreatment of adoptive transfer recipient mice abrogates the ability of their splenocytes to perpetuate passive protection in sequential adoptive transfer experiments.
Asunto(s)
Ciclosporina/administración & dosificación , Etopósido/administración & dosificación , Leucemia L1210/tratamiento farmacológico , Animales , Sinergismo Farmacológico , Inmunidad/efectos de los fármacos , Inmunización Pasiva , Leucemia L1210/inmunología , Ratones , Bazo/citología , Análisis de SupervivenciaRESUMEN
It has recently been shown that anthracycline antibiotic-resistant tumor cells are less responsive to daunorubicin-stimulated B23 nucleolar phosphoprotein translocation than drug-sensitive cells. Since cyclosporin A and verapamil reverse primary acquired and secondary cross-resistance to daunorubicin, we investigated the effect of these agents on nucleolar B23 translocation in sensitive and resistant tumors. We compared modified to baseline B23 phosphoprotein distribution between predominantly nucleolar, mixed nucleolar-nuclear, or nuclear immunofluorescence using a monoclonal anti-B23 antibody in parental drug-sensitive and multidrug-resistant acute lymphatic leukemia and in daunorubicin-sensitive and -resistant murine hepatoma. Our experiments show that cyclosporin A and verapamil alone have no effect on B23 phosphoprotein translocation, but that the addition of either agent to sensitive parental or resistant tumor sublines markedly enhances daunorubicin-stimulated translocation. This effect correlates with the correction of impaired daunorubicin inhibition of RNA synthesis by cyclosporin A and verapamil in the resistant sublines. Our observations suggest that nucleolar B23 phosphoprotein is an important site in the modulation of anthracycline antibiotic antitumor activity.
Asunto(s)
Ciclosporinas/farmacología , Daunorrubicina/farmacología , Neoplasias Hepáticas Experimentales/metabolismo , Proteínas Nucleares/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Verapamilo/farmacología , Animales , Línea Celular , Resistencia a Medicamentos , Humanos , Ratones , NucleofosminaRESUMEN
Trifluoperazine (TFP) shows cytotoxic activity against human acute lymphatic leukemia (ALL) in vitro. This activity is inhibited by increasing serum concentration and by albumin. Despite its in vitro activity, the drug is inactive in vivo. To determine if increased phenothiazine hydrophilicity could protect against albumin inhibition of antileukemic activity, we compared ALL cytotoxic median effective dose concentrations of a series of hydroxylated phenothiazines in 5% fetal bovine serum (FBS) and in 5% FBS supplemented with albumin. Albumin inhibits the activity of all drugs. A representative derivative 7,8-dihydroxychlorpromazine, although active in vitro, is inactive against L1210 and P388 murine leukemias in vivo.
Asunto(s)
Albúminas/farmacología , Leucemia/tratamiento farmacológico , Fenotiazinas/farmacología , Calmodulina/antagonistas & inhibidores , Supervivencia Celular/efectos de los fármacos , Humanos , Hidroxilación , Fenotiazinas/antagonistas & inhibidores , Relación Estructura-ActividadRESUMEN
An 18-year-old woman with severe pancytopenia secondary to chemotherapy given in the third trimester of pregnancy and who delivered an infant with normal peripheral blood counts is reported. The literature is reviewed, and recommendations for the method of delivery in this setting are discussed.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Complicaciones Neoplásicas del Embarazo , Sarcoma de Ewing/complicaciones , Adolescente , Recuento de Células Sanguíneas , Femenino , Sangre Fetal , Humanos , Recién Nacido , Pancitopenia/inducido químicamente , Embarazo , Tercer Trimestre del EmbarazoRESUMEN
We have previously developed a daunorubicin resistant subline of Ehrlich ascites carcinoma (EA/DR) for studies on the reversal of daunorubicin resistance. The mean survival of untreated BALB/c mice bearing drug sensitive parental tumour (EA/DS) is 18.4 +/- 0.6 days, mice bearing EA/DS treated with five daily doses of 0.3 mg kg-1 daunorubicin greater than 60 days, and mice bearing EA/DR treated with the same daunorubicin regimen, 21.1 +/- 1.4 days. We now report complete reversal of daunorubicin resistance in EA/DR by cyclosporin A (CsA). The in vitro daunorubicin IC50, defined as that concentration of daunorubicin required to inhibit 50% of DNA synthesis, in EA/DR was 6.7 +/- 1.15 micrograms ml-1 compared to 2.8 +/- 0.72 micrograms ml-1 in EA/DS. This value was reduced to 2.8 +/- 0.52 and 2.1 +/- 0.10 micrograms ml-1 daunorubicin by 3.3 and 13.2 micrograms ml-1 CsA respectively, P less than 0.05. The MST of groups of host mice bearing EA/DR either untreated, treated with five daily doses of 0.3 mg kg-1 daunorubicin, treated with 80 mg kg-1 CsA in five divided daily doses or treated with combined daunorubicin-CsA were 19.0 +/- 1.0, 21.1 +/- 1.4, 24.0 +/- 2.6 and greater than 60 days respectively. The mean survival of groups of host mice bearing EA/DR treated with 5 mg kg-1 or 10 mg kg-1 CsA simultaneously with daunorubicin for five days was also greater than 60 days. These differences are highly significant.
Asunto(s)
Carcinoma de Ehrlich/tratamiento farmacológico , Ciclosporinas/uso terapéutico , Daunorrubicina/uso terapéutico , Animales , Carcinoma de Ehrlich/mortalidad , Interacciones Farmacológicas , Resistencia a Medicamentos , Técnicas In Vitro , Ratones , Ratones Endogámicos BALB CRESUMEN
The development of drug resistance by tumor cells is a major obstacle to the cure of human malignancy. Cyclosporin A (CsA) completely reverses primary resistance to vincristine and cross resistance to daunorubicin in a pleiotropic drug-resistant subline of human T cell acute lymphatic leukemia. This subline is over 50-fold resistant to vincristine and fivefold resistant to daunorubicin. CsA has little effect on vincristine or daunorubicin activity in drug-sensitive parental leukemia and corrects daunorubicin resistance without altering cellular daunorubicin accumulation.
Asunto(s)
Ciclosporinas/uso terapéutico , Daunorrubicina/uso terapéutico , Leucemia Linfoide/tratamiento farmacológico , Vincristina/uso terapéutico , Línea Celular , Resistencia a Medicamentos , Sinergismo Farmacológico , Técnicas In VitroRESUMEN
Verapamil, the calcium-influx-blocking agent, has previously been shown to have favorable interactions with antineoplastic drugs. Our study of human T cell acute lymphatic leukemia (ALL) GM3639 indicates that verapamil enhances the in vitro cytotoxicity of VP-16-213 against drug-sensitive ALL by reducing the concentration of VP-16-213, resulting in 50% cell viability from 104.5 +/- 26.6 nM to 46.0 +/- 2.7 nM (P less than 0.05). The addition of verapamil to VP-16-213 treatment of BDF/1 mice bearing L1210 leukemia increases their mean survival from 21.2 +/- 3.6 to 50.4 +/- 4.3 days (P less than 0.01) and the survival of CD2F/l mice bearing P388 leukemia from 27.8 +/- 3.7 to 49.1 +/- 5.0 days (P less than 0.01). The 30-day survival is significantly increased in L1210 and P388 leukemia mice, and 60-day survival is significantly increased in P388 leukemic mice by verapamil. We developed a vincristine (VCR)-resistant subline of GM3639 T cell ALL, L23, by continuous exposure of drug-sensitive cells to VCR. This subline demonstrates pleiotropic cross resistance to VP-16-213 and daunorubicin. The addition of verapamil to VCR, to VP-16-213, and to daunorubicin completely restores responsiveness to these drugs, as indicated by the normalization of the VCR and VP-16-213 concentrations required for cytotoxicity and the concentration of daunorubicin required for inhibition of thymidine incorporation.