RESUMEN
PURPOSE: Since the role of resistin was evaluated only in patients with non-small cell lung cancer (NSCLC) not treated with immunotherapy, we aimed to evaluate levels of resistin during immunotherapy (nivolumab) and its prognostic role with regard to OS. METHODS/PATIENTS: From a cohort of 78 patients with advanced NSCLC enrolled in a prospective study at Ospedale Policlinico San Martino in Genoa (Italy), 43 patients have been considered for this sub-analysis because of the availability of samples. Before and during nivolumab administration, clinical information and blood samples were collected and resistin, matrix metalloproteinase (MMP)-8, MMP-9, and myeloperoxidase were evaluated by enzyme-linked immunosorbent assay (ELISA). RESULTS: Median age was 71 with a prevalence of males and former smokers. Median resistin levels presented a peak at cycle 2 and then dropped down until the last cycle. Resistin correlated with all neutrophil degranulation products at cycle 1 (except for MMP-9) and at cycle 2 as well as with white blood cells and neutrophils. By a ROC curve analysis, a resistin value at cycle 2 of 19 ng/mL was tested as the best cut-off point for OS. Kaplan-Meier analysis demonstrated that patients above the resistin cut-off experienced a reduced OS (median OS 242.5 vs. 470 days, p = 0.0073), as confirmed by Cox proportional hazards regression analysis. CONCLUSIONS: Resistin levels > 19 ng/mL at the time of the second cycle of nivolumab treatment independently predict a reduced OS in patients with advanced NSCLC.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/sangre , Neoplasias Pulmonares/sangre , Nivolumab/uso terapéutico , Resistina/sangre , Anciano , Antineoplásicos Inmunológicos/uso terapéutico , Biomarcadores de Tumor/sangre , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/inmunología , Carcinoma de Pulmón de Células no Pequeñas/mortalidad , Femenino , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/mortalidad , Masculino , Persona de Mediana Edad , Neutrófilos/citología , Pronóstico , Estudios Prospectivos , Tasa de Supervivencia , Resultado del TratamientoRESUMEN
BACKGROUND: CEA, CYFRA21-1 and NSE are tumor markers used for monitoring the response to chemotherapy in advanced adenocarcinoma, squamous cell carcinoma and small-cell lung cancer, respectively. Their role in cancer immunotherapy needs to be elucidated. METHODS: Patients with advanced non-small cell lung cancer (NSCLC) were treated with nivolumab 3 mg/kg every 2 weeks within the Italian Nivolumab Expanded Access Program. Blood samples were collected at baseline, at each cycle up to cycle 5 and then every two cycles until patient's withdrawn from the study. All patients underwent a CT-scan after every 4 cycles of treatment and responses were classified according to RECIST 1.1. The biomarkers serum levels were measured with a chemiluminescent microparticle immunoassay for CEA and with an immuno radiometric assay for CYFRA21-1 and NSE. The markers values at baseline and after 4 cycles were used to analyze the relationship between their variation over baseline and the tumor response, evaluated as disease control rate (DCR: CR + PR + SD), and survival (PFS and OS). RESULTS: A total of 70 patients were evaluable for the analysis. Overall, a disease control was obtained in 24 patients (35.8%, 4 PR + 20 SD). After 4 cycles of nivolumab a CEA or CYFRA21-1 reduction ≥ 20% over the baseline was significantly associated with DCR (CEA, p = 0.021; CYFRA21-1, p < 0.001), PFS (CEA, p = 0.028; CYFRA21-1, p < 0.001) and OS (CEA, p = 0.026; CYFRA21-1, p = 0.019). Multivariate analysis confirmed the ability of CYFRA21-1 reduction ≥ 20% to predict DCR (p = 0.002) and PFS (p < 0.001). CONCLUSION: The reduction in serum level of CYFRA21-1 or CEA might be a reliable biomarker to predict immunotherapy efficacy in NSCLC patients. NSE was not significant for monitoring the efficacy of nivolumab.
Asunto(s)
Antígenos de Neoplasias/sangre , Antígeno Carcinoembrionario/sangre , Carcinoma de Pulmón de Células no Pequeñas/sangre , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Queratina-19/sangre , Neoplasias Pulmonares/sangre , Neoplasias Pulmonares/tratamiento farmacológico , Nivolumab/uso terapéutico , Fosfopiruvato Hidratasa/sangre , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/sangre , Carcinoma de Pulmón de Células no Pequeñas/patología , Supervivencia sin Enfermedad , Femenino , Humanos , Neoplasias Pulmonares/patología , Masculino , Persona de Mediana Edad , Estadificación de NeoplasiasRESUMEN
INTRODUCTION: Vinflunine belongs to the class of vinca alkaloids and acts by disrupting the microtubule dynamics during cell cycle; this agent is currently available for previously treated advanced transitional cell carcinoma in Europe. The aim of this invited review is to evaluate the potential role of vinflunine for the treatment of non-small cell lung cancer (NSCLC). Areas covered: The potential role of vinflunine in NSCLC is discussed on the basis of the available data, including full papers and meeting abstracts. Relevant preclinical studies describing the pharmacological properties of vinflunine are also included. The review also summarizes clinical studies, including phase I trials involving NSCLC among other tumors as well as phase II/III trials specifically addressing this malignancy. Additionally, the safety profile and the current regulatory status of vinflunine is discussed. Expert opinion: Vinflunine is active as single agent and as part of platinum-based combinations in NSCLC. It results non-inferior to docetaxel in a randomized phase III trial including previously treated NSCLC patients; additionally, its safety profile is generally considered manageable. Ultimately, further studies are needed to confirm the role of vinflunine in NSCLC, in consideration of the evolving evidence regarding targeted therapies and immune check-point inhibitors.
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Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Neoplasias Pulmonares/tratamiento farmacológico , Vinblastina/análogos & derivados , Animales , Antineoplásicos Fitogénicos/administración & dosificación , Antineoplásicos Fitogénicos/farmacología , Carcinoma de Pulmón de Células no Pequeñas/patología , Humanos , Neoplasias Pulmonares/patología , Ensayos Clínicos Controlados Aleatorios como Asunto , Vinblastina/administración & dosificación , Vinblastina/efectos adversos , Vinblastina/farmacologíaRESUMEN
Preclinical studies have suggested that combining cytotoxic agents with epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) to treat EGFR-mutated tumors may increase their inhibitory effect depending on the order of drug administration. The antitumor efficacy of different treatment sequences using vinorelbine (VNB) and gefitinib (GEF) was investigated both in vitro and in vivo in non-small cell lung cancer (NSCLC) cell lines with the rationale of potentially translating these findings into the clinical setting. The EGFR-wild-type A549 and the EGFR-mutated (exon 21 L858R/exon 20 T790M) H1975 cell lines were treated as follows: GEF followed by VNB, VNB followed by GEF and the two drugs applied individually or concurrently. Results in vitro demonstrated that the sequence of VNB followed by GEF was significantly more active than single-agent treatments. The expression of activated EGFR and its downstream pathway genes indicated that the increased cytotoxic effect of the VNB and GEF treatment sequence was accompanied by inhibition of EGFR, AKT and ERK1/2. Moreover, the increased inhibition of tumor growth after treatment with VNB followed by GEF was also confirmed in CD1-nude mice that were xenotransplanted with H1975 cells (p < 0.0001). This effect was paralleled by a corresponding decrease in cancer glucose consumption, as assessed by micro-positron emission tomography scans (p < 0.05). These preclinical findings in NSCLC cell lines, which are poorly responsive to EGFR-TKIs, demonstrated that the sequential treatment of VNB followed by GEF induced a significant antitumor effect, which supports the translation of this treatment schedule into a clinical setting.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Neoplasias Pulmonares/tratamiento farmacológico , Animales , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carcinoma de Pulmón de Células no Pequeñas/diagnóstico por imagen , Línea Celular Tumoral , Esquema de Medicación , Resistencia a Antineoplásicos , Receptores ErbB/antagonistas & inhibidores , Receptores ErbB/metabolismo , Femenino , Fluorodesoxiglucosa F18/farmacocinética , Gefitinib , Humanos , Neoplasias Pulmonares/diagnóstico por imagen , Ratones Desnudos , Inhibidores de Proteínas Quinasas/farmacología , Quinazolinas/administración & dosificación , Cintigrafía , Radiofármacos/farmacocinética , Transducción de Señal , Vinblastina/administración & dosificación , Vinblastina/análogos & derivados , Vinorelbina , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Malignant mesothelioma (MM) is an aggressive tumor, mainly derived from the pleura, which is predominantly associated with exposure to asbestos fibers. The prognosis of MM patients is particularly severe, with a median survival of approximately 9-12 months and latency between exposure and diagnosis ranging from 20-50 years (median 30 years). Emerging evidence has demonstrated that tumor aggressiveness is associated with genome and gene expression abnormalities; therefore, several studies have recently focused on the role of microRNAs (miRNAs) in MM tumorigenesis. miRNAs are small non-protein coding single-stranded RNAs (17-22 nucleotides) involved in numerous cellular processes that negatively regulate gene expression by modulating the expression of downstream target genes. miRNAs are often deregulated in cancer; in particular, the differential miRNA expression profiles of MM cells compared to unaffected mesothelial cells have suggested potential roles of miRNAs as either oncogenes or tumor suppressor genes in MM oncogenesis. In this review, the mechanism of MM carcinogenesis was evaluated through the analysis of the published miRNA expression data. The roles of miRNAs as diagnostic biomarkers and prognostic factors for potential therapeutic strategies will be presented and discussed.
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Regulación Neoplásica de la Expresión Génica , Neoplasias Pulmonares/genética , Mesotelioma/genética , MicroARNs/genética , Animales , Biomarcadores de Tumor/genética , Humanos , Pulmón/metabolismo , Pulmón/patología , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/terapia , Mesotelioma/diagnóstico , Mesotelioma/terapia , Mesotelioma Maligno , Oncogenes , PronósticoRESUMEN
The effects of different marine derived agents were studied in A549 cell growth. These drugs induced cell cycle arrest at the G2-M phase associated with the up-regulation of GADD45alpha-gamma and down-regulation of c-Myc. In treated cells, GADD45alpha-gamma and c-Myc were up- and down-regulated, respectively. A cascade of events leading to apoptotic mitochondrial 'intrinsic' pathway was observed in treated cells: (1) dephosphorylation of BAD serine136; (2) BAD dissociation from 14-3-3 followed by its association with BCL-XL; (3) cytochrome c release; (4) caspase-3 activation, and (5) cleavage of vimentin. Caspase(s) inhibitor prevented the formation of cleavage products and, in turn, apoptosis was inhibited through a p53-independent mechanism. Moreover, these compounds did not activate NF-kappaB. Our findings may offer new insights into the mechanisms of action of these agents in A549 cells. The better understanding of their effects might be important to fully exploit the potential of these new drugs.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis , Toxinas Marinas/farmacología , Proteínas 14-3-3/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Ciclo Celular/efectos de los fármacos , Ciclo Celular/fisiología , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Línea Celular Tumoral , Citocromos c/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Neoplasias Pulmonares/patología , Mitocondrias/metabolismo , FN-kappa B/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fosforilación , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , Vimentina/metabolismo , Proteína Letal Asociada a bcl/metabolismo , Proteína bcl-X/metabolismoRESUMEN
Organotin compounds, particularly tri-organotin, have demonstrated cytotoxic properties against a number of tumor cell lines. On this basis, triethyltin(IV)lupinylsulfide hydrochloride (IST-FS 29), a quinolizidine derivative, was synthesized and developed as a potential antitumor agent. This tin-derived compound exhibited potent antiproliferative effects on three different human cancer cell lines: teratocarcinoma of the ovary (PA-1), colon carcinoma (HCT-8) and glioblastoma (A-172). Cytotoxic activity was assessed by MTT and cell count assays during time course experiments with cell recovery after compound withdrawal. Significant cell growth inhibition (up to 95% in HCT-8 after 72 h of exposure), which also persisted after drug-free medium change, was reported in all the cell lines by both assays. In addition, the cytocidal effects exerted by IST-FS 29 appeared more consistent with necrosis or delayed cell death, rather than apoptosis, as shown by morphologic observations under light microscope, DNA fragmentation analysis and flow cytometry. In the attempt to elucidate whether this compound might affect genes playing a role in G1/S phase transition, the expressions of p53, p21(WAF1), cyclin D1 and Rb, mainly involved in response to DNA-damaging stress, were analyzed by Western blot. Heterogeneous patterns of expression during exposure to IST-FS 29 were evidenced in the different cell lines suggesting that these cell-cycle-related genes are not likely the primary targets of this compound. Thus, the present data seem more indicative of a direct effect of IST-FS-29 on macromolecular synthesis and cellular homeostasis, as previously hypothesized for other organotin complexes.
Asunto(s)
Antineoplásicos/farmacología , Proteínas de Ciclo Celular/metabolismo , Neoplasias del Colon/tratamiento farmacológico , Glioblastoma/tratamiento farmacológico , Neoplasias Ováricas/tratamiento farmacológico , Teratocarcinoma/tratamiento farmacológico , Compuestos de Trietilestaño/farmacología , Antineoplásicos/metabolismo , Western Blotting , Proteínas de Ciclo Celular/efectos de los fármacos , División Celular/efectos de los fármacos , Neoplasias del Colon/metabolismo , Neoplasias del Colon/patología , Fragmentación del ADN/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Citometría de Flujo , Glioblastoma/metabolismo , Glioblastoma/patología , Humanos , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/patología , Teratocarcinoma/metabolismo , Teratocarcinoma/patología , Compuestos de Trietilestaño/metabolismo , Células Tumorales CultivadasRESUMEN
OBJECTIVE: To identify through a substudy of a larger, multicenter study of adjuvant treatment in primary operable breast cancer patients any possible correlation between cellular proliferation rate, measured by thymidine labeling index (TLI), and perioperative chemotherapy (periCT). METHODS: TLI was measured in slices of early breast carcinoma patients. The main trial was designed to randomize patients after primary surgery to receive one cycle of periCT consisting of cyclophosphamide, epidoxorubicin and 5-fluorouracil, or no periCT. RESULTS: Of 600 patients randomized into the main study, 197 were eligible for inclusion in this substudy. Characteristics of patients were quite similar to those of the entire population entered into the main study. The TLI cutoff value in our series was 0.7% expressed as the median percentage ratio of thymidine-labeled cells undergoing DNA synthesis in the tumor cell population of specimens from the 197 patients. No differences were observed in terms of relapse-free survival (RFS) and overall survival (OS) after grouping the patients by TLI value (low and high) and by treatment. Among node-negative patients, a significant improvement in terms of OS (p = 0.02), but not RFS (p = 0.06), was seen in patients with a high-TLI value who underwent periCT versus controls. CONCLUSIONS: TLI may be a useful tool for the identification of node-negative patients with high-TLI values who may benefit from periCT.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias de la Mama/diagnóstico , Neoplasias de la Mama/tratamiento farmacológico , Timidina , Adulto , Neoplasias de la Mama/patología , Neoplasias de la Mama/cirugía , Quimioterapia Adyuvante , Femenino , Humanos , Persona de Mediana Edad , Ensayos Clínicos Controlados Aleatorios como Asunto , Riesgo , Factores de Riesgo , Análisis de SupervivenciaRESUMEN
Indole-3-carbinol is a natural product which has been shown to reduce the incidence of spontaneous and carcinogen-induced mammary tumours in animals. Eighteen unsymmetrical methylene derivatives of indoles were prepared by reaction of Mannich bases of 7-hydroxycoumarins with substituted indoles in acetic or propionic anhydride. The synthesised molecules were tested in vitro against the MCF7 and MDA-MB-231 breast cancer cell lines by MTT and cell count assays. Results from 16 tested compounds showed that 60% of them exerted some effects against the MDA-MB-231 compared to about 30% towards the MCF7. Among all, the 3-(7'-acetoxy-4-methylcoumarin-8'-yl)methyl-2-methylindole resulted the most effective in both cell lines, compared to indole-3-carbinol. In conclusion, these preliminary results report that some of these compounds might be promising potential antiproliferative agents.
Asunto(s)
Antineoplásicos/síntesis química , Cumarinas/síntesis química , Indoles/síntesis química , Adenocarcinoma , Antineoplásicos/química , Antineoplásicos/farmacología , Neoplasias de la Mama , División Celular/efectos de los fármacos , Cumarinas/química , Cumarinas/farmacología , Humanos , Indoles/química , Indoles/farmacología , Células Tumorales CultivadasRESUMEN
The cytotoxic effect and antitumor activity induced by the novel organotin compound triethyltin(IV)lupinyisulfide hydrochloride, have been investigated. Different patterns of antiproliferative effects have been observed in a panel of human tumor cell lines in vitro. Toxicity studies in mice reported acute toxicity at the doses of 21 and 17.5 mg/kg which progressively disappeared at lower concentrations. On this basis, the doses of 3.5, 7 and 14 mg/kg were selected to assess the antitumor activity in vivo against the P388 leukemic cells xenografted in mice. This compound was able to induce a dose-dependent significant reduction of tumor volume, up to 46%, at the highest concentration (p = 0.0062) without important toxicity, as also confirmed by histological analysis of the main organ tissues. This preliminary study seems to hold interest for further investigations in different tumor models as well as for the evaluation of optimal drug route and schedule.
Asunto(s)
Antineoplásicos/toxicidad , Supervivencia Celular/efectos de los fármacos , Riñón/patología , Leucemia P388/tratamiento farmacológico , Compuestos Orgánicos de Estaño/toxicidad , Compuestos de Trietilestaño/toxicidad , Animales , Antineoplásicos/uso terapéutico , Femenino , Humanos , Células Jurkat , Riñón/efectos de los fármacos , Leucemia P388/patología , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Compuestos Orgánicos de Estaño/uso terapéutico , Trasplante Heterólogo , Compuestos de Trietilestaño/uso terapéutico , Células Tumorales CultivadasRESUMEN
Alterations in the expression of cyclin D1 have been reported frequently in several human cancers, but their significance in the multistep model of carcinogenesis has been scantly described. To define the pattern of cyclin D1 expression in the development of ovarian cancer and clinical outcome, 55 cases of benign ovarian tumors, 12 borderline cases, and 37 ovarian carcinomas (32 primary and 5 recurrent carcinomas) were studied. Analyses were carried out on fresh tumor specimens by Western blotting and reverse transcription-PCR and provided significant superimposable results (P = 0.00001). Cyclin D1 abundance was classed according to the densitometric values as undetectable, detectable, well detectable, and highly detectable. A significant increase (P < 0.000001) in median cyclin D1 values was observed from benign (0.038; range, 0.001-0.705) to borderline (0.226; range, 0.001-0.623) to malignant (0.347; range, 0.027-2.330) to recurrent (0.887; range, 0.309-2.2260) tumors. In addition, higher median cyclin D1 values were reported in serous carcinomas (P = 0.058) and advanced-stage diseases (P = 0.003). Survival analyses carried out in the 32 primary carcinomas showed no significant difference in overall survival between detectable versus well/highly detectable cyclin D1 neoplasms. Conversely, a significant relationship between cyclin D1 expression and progression-free survival was found (P = 0.031). These results may elucidate the function of altered cyclin D1 expression in ovarian tumorigenesis and provide a basis for additional studies on its prognostic role.
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Biomarcadores de Tumor/biosíntesis , Ciclina D1/biosíntesis , Neoplasias Ováricas/metabolismo , Femenino , Humanos , Persona de Mediana Edad , Metástasis de la Neoplasia , Recurrencia Local de Neoplasia , Neoplasias Ováricas/mortalidad , Neoplasias Ováricas/patología , Tasa de SupervivenciaRESUMEN
We have investigated the patterns of in vitro cytotoxicity, induced by six newly synthesized gold and tin compounds, in three human ovarian cancer cell lines (SW 626, IGROV 1 and OVCAR-3). Four gold compounds, i.e. gold(I)lupinylsulfide hydrochloride [1] (containing a naked gold atom), triethylphosphinogold(I)lupinylsulfide hydrochloride [2], triphenyl-phosphinogold(I)lupinylsulfide hydrochloride [3] and 1 ,2-bis(diphenylphosphino)ethane bis[gold(I)lupinylsulfide] dihydrochloride [4] (all containing a gold atom coordinated with different phosphines), were prepared. Moreover, the triethylphosphinogold(I)(2-diethylamino)ethylsulfide hydrochloride [5] in which the simple diethylaminoethylthiol replaced the bulky lupinylthiol was synthesized. The tin compound, triethyltin(IV)lupinylsulfide hydrochlorlde [6], was also studied. Comparative tests with cisplatin, the most widely used antitumor agent in ovarian cancer, were carried out in biological Investigations. In vitro cytotoxicity, by MTT assay, showed that compound [4] and compound [6] exhibited interesting antiproliferative activity in all the three cell lines (mean IC50=1.3 and 0.7 microM, respectively) compared to cisplatin (mean IC50=4.8 microM). In addition, the PA-1 cell line, more sensitive to cisplatin (IC50=0.6 microM), was included as a comparison in the study. Cell count assays confirmed the cytotoxic properties of compounds [4] and [6] against the four cell lines, reporting higher growth Inhibition potency than cisplatin, with IC50 values in the sub-micromolar range.
Asunto(s)
Antineoplásicos/uso terapéutico , Compuestos de Oro/uso terapéutico , Neoplasias Ováricas/tratamiento farmacológico , Compuestos de Estaño/uso terapéutico , Antineoplásicos/química , Cisplatino/uso terapéutico , Relación Dosis-Respuesta a Droga , Femenino , Compuestos de Oro/química , Humanos , Técnicas In Vitro , Compuestos de Estaño/química , Células Tumorales Cultivadas/metabolismoRESUMEN
Cyclin D1 is a critical gene controlling the G1 phase progression through the cell cycle. Alterations of cyclin D1 have been demonstrated in a variety of cancer types. We recently reported that increased cyclin D1 expression is associated with malignancy also in ovarian tumors. Three human ovarian cancer cell lines (SW626, OVCAR-3, IGROV1), expressing high levels of this gene, were used to investigate the effects induced by antisense oligonucleotides to cyclin D1 as antiproliferative compounds. Unmodified 18 mer oligomers, targeted to the translation start site of the cyclin D1 cDNA, were able to inhibit the growth of the three cell lines after a single administration of 40 microM. The pattern of cell number reduction ranged between 30 and 55% after 48 h of treatment. Moreover, by RT-PCR and Western blotting, a marked decrease of the cyclin D1 transcript and protein (up to 77% in the SW626) was detected after 24 and 48 h, respectively, from antisense exposure. Conversely, no relevant inhibition was reported in the sense-treated cells. The present data confirm the role of cyclin D1 expression in the proliferative behavior of ovarian cancer and provide additional information that might be helpful in the search for new therapeutic strategies of this disease.
Asunto(s)
Regulación Neoplásica de la Expresión Génica , Genes bcl-1 , Oligonucleótidos Antisentido/metabolismo , Neoplasias Ováricas/metabolismo , Western Blotting , Cartilla de ADN , Femenino , Humanos , Neoplasias Ováricas/genética , Reacción en Cadena de la Polimerasa/métodos , ADN Polimerasa Dirigida por ARN , Células Tumorales Cultivadas/metabolismoRESUMEN
Cyclin D1 is a cell cycle regulator of G1 progression that has been suggested to play a relevant role in the pathogenesis of several human cancer types. In the current study, the expression of cyclin D1 has been investigated in a series of 33 patients, with benign (10 patients), borderline (five patients) and malignant (18 patients) ovarian disease. Cyclin D1 protein and mRNA content were analysed by Western blotting and reverse transcriptase polymerase chain reaction respectively. The levels of cyclin D1 protein were undetectable in patients with benign disease, detectable in the majority of patients with borderline disease and elevated in those with ovarian carcinomas, being significantly related to the degree of malignancy (carcinoma vs benign, P = 0.0001; benign vs borderline, P = 0.0238). A significant relationship between cyclin D1 expression and tumour proliferative activity was also found (P = 0.000001). Moreover, eight benign lesions, two borderline tumours and 11 carcinomas proved to be suitable for the analysis of cyclin D1 transcript, and emerging data demonstrated significant agreement between protein abundance and mRNA expression. Results from the current study suggest that cyclin D1 expression is associated with the degree of transformation and most probably plays a role in the early development of ovarian malignancy.
Asunto(s)
Carcinoma/metabolismo , Ciclinas/biosíntesis , Proteínas Oncogénicas/biosíntesis , Neoplasias Ováricas/metabolismo , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Western Blotting , Carcinoma/etiología , Carcinoma/patología , División Celular , Transformación Celular Neoplásica/metabolismo , Transformación Celular Neoplásica/patología , Ciclina D1 , Ciclinas/genética , Cartilla de ADN/química , Femenino , Humanos , Persona de Mediana Edad , Proteínas Oncogénicas/genética , Neoplasias Ováricas/etiología , Neoplasias Ováricas/patología , Reacción en Cadena de la Polimerasa , ARN Mensajero/metabolismo , Timidina/metabolismo , Células Tumorales CultivadasRESUMEN
The potential for modulating gene expression by the use of antisense oligonucleotides has become increasingly interesting in recent years. Antisense oligonucleotides are complementary nucleic acid fragments that hybridize to target sequences within RNA to form a DNA-RNA duplex, resulting in the block of translation of messenger RNA into the protein. Advances in chemistry and molecular biology have provided the basis to develop antisense oligodeoxynucleotides and improve their selectivity, stability and specificity of action. The antisense technology has been extensively used in vitro and in vivo as a tool to study the regulatory mechanisms in biologic processes and as potential therapeutic agents in cancer, viral infections and genetic disorders. In the present review, the various approaches for the use of antisense molecules in oncology, virology, genetic and inflammatory diseases are described; several studies, supporting the in vitro and in vivo applications of this technology, are also presented. Moreover, the potential clinical use of antisense therapies is discussed.
Asunto(s)
Terapia Genética , Oligonucleótidos Antisentido/uso terapéutico , Antineoplásicos/uso terapéutico , Antivirales/uso terapéutico , Enfermedades Genéticas Congénitas/terapia , Humanos , Neoplasias/terapia , Oligonucleótidos Antisentido/farmacocinética , Virosis/terapiaRESUMEN
Three human cancer cell lines (OC 314, OC 315, and OC 316) were newly established in permanent culture from the ascites of patients with serous adenocarcinoma of the ovary. OC 314 was derived from an untreated tumor presenting with ascites at diagnosis; OC 315 was isolated from a neoplasm progressing after cisplatin-containing regimen; and OC 316 was collected from a patient with pleural metastasis at diagnosis, resistant to different chemotherapeutic treatments including Taxol. These cell lines were repetitively subcultured once to twice a week through 75-80 passage generations. Tumor cells grew as monolayers and displayed epithelial-like morphology, consistent with a feature of adenocarcinoma, which was then confirmed by the expressions of cytokeratins and vimentin. The cell lines proved highly tumorigenic when transplanted into nude mice, both subcutaneously and intraperitoneally. In addition, the mice inoculated with subcutaneous OC 316 developed extremely aggressive tumor, also invading the peritoneum, which correlated with the malignant behavior of the original tumor. Drug sensitivity, evaluated by the MTT assay, showed that the three cell lines expressed similar sensitivity to doxorubicin. Responses to cisplatin essentially reported low sensitivity of OC 314 and OC 315 and resistance of OC 316, thus reflecting the original sensitivity at the clinical level.
Asunto(s)
Ascitis/patología , Cistadenocarcinoma Seroso/patología , Neoplasias Ováricas/patología , Células Tumorales Cultivadas , Animales , División Celular , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Citometría de Flujo , Humanos , Inmunohistoquímica , Cinética , Ratones , Ratones DesnudosRESUMEN
The antiproliferative effects mediated by a 14-mer homopyrimidine oligonucleotide (5' CTTTCT-CTTTTCTC3'), designed to form DNA triplex with a purine region of the DNA polymerase alpha promoter, were evaluated on the human breast cancer cell line MDA-MB 231. In order to stabilize the triple complex under physiologic conditions, replacement of cytosines by methylcytosines in the oligomer sequence was carried out. Band-shift analyses demonstrated a complete triplex formation between the radiolabeled target duplex DNA and the methylcytosine-modified oligomer at the concentration of 0.1 microM under physiologic pH and temperature. A single exposure of MDA-MB 231 cells to 0.5 microM methylcytosine-modified oligonucleotide was able to markedly reduce the cell number and the percentage of cells in DNA synthesis up to 58% and 66%, respectively, compared with controls. Furthermore, a 48% reduction in the amount of the DNA polymerase alpha mRNA was reported after treatment with the oligomer. In conclusion, data from the present study demonstrate that an oligonucleotide to DNA polymerase alpha promoter, designed to form a triple helix with target double-stranded DNA, inhibits the expression of the reporter gene at the biologic and molecular levels, suggesting a possible triplex-mediated mechanism of action.
Asunto(s)
ADN Polimerasa II/efectos de los fármacos , Oligonucleótidos/farmacología , Secuencia de Bases , Neoplasias de la Mama/patología , División Celular/efectos de los fármacos , ADN Polimerasa II/biosíntesis , ADN Polimerasa II/genética , Femenino , Humanos , Datos de Secuencia Molecular , Oligonucleótidos/genética , Unión Proteica , Pliegue de Proteína , Células Tumorales CultivadasRESUMEN
Most human follicular B-cell lymphomas are associated with t(14;18) chromosome translocation that joins the bcl-2 gene with the IgH locus. This hybrid gene causes upregulation of BCL-2 protein expression, endowing cells with survival advantage. Although early BCL-2 overexpression is definitely responsible for immortalization/transformation, its exact role in the overt transformation as well as in the maintenance of the tumor phenotype is not known. The capacity of oligodeoxynucleotides (ODN) to modulate gene expression specifically has been exploited to downregulate the overexpression of BCL-2 protein in the SU-DHL-4 human follicular B-cell lymphoma line by the use of sense ODN or antisense ODN or antisense ODN designed to encompass the unique nucleotide sequence in the fusion region of the hybrid transcript. The specific downregulation of the bcl-2 transcript and of the relevant BCL-2 protein in the treated cells activated programed cell death and inhibited growing cells. The antitumor activity was restricted to the DHL-4 cell line carrying the specific nucleotide sequence at the bcl-2/IgH joining region. Thus, DHL-4 lymphoma cells derived from the acute phase of human follicular B-cell lymphoma, although endowed with additional activated oncogenes, were growth inhibited by bcl-2 downregulation with additional activated oncogenes, were growth inhibited by bcl-2 downregulation in a genetically restricted fashion. The biological activity was exerted exclusively by ODNs synthesized in the sense orientation. The sense ODNs have been proposed to anneal the hybrid bcl-2/IgH antisense RNA as identified in this study.
Asunto(s)
Apoptosis , Proteínas Proto-Oncogénicas/genética , Secuencia de Aminoácidos , Secuencia de Bases , Cromosomas Humanos Par 14 , Cromosomas Humanos Par 18 , Daño del ADN , Cartilla de ADN/química , Regulación Neoplásica de la Expresión Génica , Genes de Inmunoglobulinas , Humanos , Datos de Secuencia Molecular , Proteínas Proto-Oncogénicas c-bcl-2 , ARN Mensajero/genética , ARN Neoplásico/genética , Translocación Genética , Células Tumorales CultivadasRESUMEN
We have investigated the possibility of inducing a kinetic recruitment of breast cancer cells by the in vivo administration of recombinant human Growth Hormone (rhGH). Twelve patients with advanced breast cancer received rhGH i.m. for 2 days immediately before the first course of chemotherapy. The following biological parameters have been evaluated before and 24 hours after rhGH administration: tumor TLI, tumor IGF-I content, serum IGF-I concentration. The mean tumor TLI values before and after rhGH were 1.3% and 2.6% respectively; median tumor and serum IGF-I levels before rhGH were 4.64 ng/g and 63.5 ng/ml respectively; after the administration of rhGH median tumor IGF-I content was 1.8 and median serum IGF-I level was 112 ng/ml. These data suggest that, in vivo, rhGH stimulates breast cancer cell proliferation; the mitogenic stimulus is likely due to the local production of IGF-I induced by rhGH.
Asunto(s)
Neoplasias de la Mama/patología , Hormona del Crecimiento/farmacología , Anciano , Neoplasias de la Mama/química , División Celular/efectos de los fármacos , Femenino , Humanos , Factor I del Crecimiento Similar a la Insulina/análisis , Persona de Mediana Edad , Proyectos Piloto , Proteínas Recombinantes/farmacologíaRESUMEN
Fine-needle aspiration (FNA) provides a suitable diagnostic tool in the management of patients with breast cancer lesions. The current study reports on tumor proliferative activity, by 3H-Thymidine Labelling Index (TLI), assessed on 59 FNA (TLI1) and 28 surgical specimens (TLI2) from the same breast cancer patients. Median TLI values from FNA and surgical material were 1.0% and 0.7%, respectively. In the 28 patients, evaluable for the comparison between TLI1 and TLI2, the association was found to be highly significant (p = 0.000). Moreover, no change in tumor proliferative activity was observed in the majority (79%) of cases when evaluated preoperatively and at surgery. This study confirms the feasibility of TLI analysis on FNA from breast cancer and provides results superimposable on those obtained in a tissue sample from the same patient.