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Jose Prados,
Consolacion Melguizo,
Gloria Perazzoli,
Laura Cabeza,
Esther Carrasco,
Jaime Oliver,
Cristina Jiménez-Luna,
Maria C Leiva,
Raúl Ortiz,
Pablo J Alvarez,
Antonia Aranega
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ABSTRACT: Gastrointestinal cancers remain one of the main causes of death in developed countries. The main obstacles to combating these diseases are the limitations of current diagnostic techniques and the low stability, availability, and/or specificity of pharmacological treatment. In recent years, nanotechnology has revolutionized many fields of medicine, including oncology. The association of chemotherapeutic agents with nanoparticles offers improvement in the solubility and stability of antitumor agents, avoidance of drug degradation, and reductions in therapeutic dose and toxicity, increasing drug levels in tumor tissue and decreasing them in healthy tissue. The use of specific molecules that drive nanoparticles to the tumor tissue represents a major advance in therapeutic specificity. In addition, the use of nanotechnology in contrast agents has yielded improvements in the diagnosis and the follow-up of tumors. These nanotechnologies have all been applied in gastrointestinal cancer treatment, first in vitro, and subsequently in vivo, with promising results reported in some clinical trials. A large number of patents have been generated by nanotechnology research over recent years. The objective of this paper is to review patents on the clinical use of nanoparticles for gastrointestinal cancer diagnosis and therapy and to offer an overview of the impact of nanotechnology on the management of this disease.
Recent patents on anti-cancer drug discovery. 05/2013;
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ABSTRACT: Colorectal cancer is the third most common cancer in both men and women and has shown a progressive increase over the past 20 years. Current chemotherapy has major limitations, and a novel therapeutic approach is required. Given that neoplastic transformation of colon epithelial cells is a consequence of genetic and epigenetic alterations, RNA interference (RNAi) has been proposed as a new therapeutic strategy that offers important advantages over conventional treatments, with high specificity and potency and low toxicity. RNAi has been employed as an effective tool to study the function of genes, preventing their expression and leading to the development of new approaches to cancer treatment. In malignancies, including colon cancer, RNAi is being used for "silencing" genes that are deregulated by different processes such as gene amplification, mutation, or overexpression and may be the cause of oncogenesis. This strategy not only provides information on the involvement of certain genes in colon cancer, but also opens up a new perspective for its treatment. However, most studies have used adenovirus or lentivirus vectors to transport RNAi into tumor cells or tumors in animal models, because several technical obstacles must be overcome before RNAi can be used in the clinical setting. The aim of this study was to review current knowledge on the use of RNAi techniques in the treatment of colon cancer.
BioDrugs 04/2013; · 3.44 Impact Factor
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ABSTRACT: Accessory (supernumerary) intrathoracic ribs are a very rare congenital disorder. Here, we present the first case of multiple supernumerary intrathoracic ribs in an adult, which are present consecutively between ribs 1 and 4 and without articulation with the vertebrae. Despite this, anatomical variation is usually silent and accidentally discovered; its knowledge can prevent confusion with other structures during imaging diagnostic techniques of thoracic pathologies.
Anatomia Clinica 02/2013; · 0.93 Impact Factor
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Consolación Melguizo, Jose Prados,
Beatriz González,
Raul Ortiz,
Angel Concha,
Pablo Juan Alvarez,
Roberto Maddedu,
Gloria Perazzoli,
Jaime Antonio Oliver,
Rodrigo López,
Fernando Rodríguez-Serrano,
Antonia Aránega
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ABSTRACT: BACKGROUND: The CD133 antigen is a marker of radio- and chemo-resistant stem cell populations in glioblastoma (GBM). The O6-methylguanine DNA methyltransferase (MGMT) enzyme is related with temozolomide (TMZ) resistance. Our propose is to analyze the prognostic significance of the CD133 antigen and promoter methylation and protein expression of MGMT in a homogenous group of GBM patients uniformly treated with radiotherapy and TMZ. The possible connection between these GBM markers was also investigated. METHODS: Seventy-eight patients with GBM treated with radiotherapy combined with concomitant and adjuvant TMZ were analyzed for MGMT and CD133. MGMT gene promoter methylation was determined by methylation-specific polymerase chain reaction after bisulfite treatment. MGMT and CD133 expression was assessed immunohistochemically using an automatic quantification system. Overall and progression-free survival was calculated according to the Kaplan--Meier method. RESULTS: The MGMT gene promoter was found to be methylated in 34 patients (44.7%) and unmethylated in 42 patients (55.3%). A significant correlation was observed between MGMT promoter methylation and patients' survival. Among the unmethylated tumors, 52.4% showed low expression of MGMT and 47.6% showed high-expression. Among methylated tumors, 58.8% showed low-expression of MGMT and 41.2% showed high-expression. No correlation was found between MGMT promoter methylation and MGMT expression, or MGMT expression and survival. In contrast with recent results, CD133 expression was not a predictive marker in GBM patients. Analyses of possible correlation between CD133 expression and MGMT protein expression or MGMT promoter methylation were negative. CONCLUSIONS: Our results support the hypothesis that MGMT promoter methylation status but not MGMT expression may be a predictive biomarker in the treatment of patients with GBM. In addition, CD133 should not be used for prognostic evaluation of these patients. Future studies will be necessary to determine its clinical utility.
Journal of Translational Medicine 12/2012; 10(1):250. · 3.41 Impact Factor
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ABSTRACT: PURPOSE: Multidrug resistance is one of the major obstacles to the successful treatment of non-small cell lung cancer (NSCLC). An ability to identify molecular markers of drug resistance in peripheral blood cells in order to better target treatment would therefore be extremely useful in selecting therapy protocols for patients. The aim of the present study was to evaluate whether expression of resistance genes (MDR1, MRP3 and LRP) can predict clinical outcome in NSCLC patients treated with paclitaxel and carboplatin. METHODS: Peripheral blood samples were obtained from lung cancer patients before and after chemotherapy and expression of the resistance gene in polymononuclear cells was detected by real-time reverse-transcription polymerase chain reaction. The results were correlated with treatment response and overall survival, which was calculated according to the Kaplan-Meier method. RESULTS: MDR1 expression levels in PMNs rose rapidly within 24 h post-administration of paclitaxel and carboplatin, whereas MRP and LRP expression levels remained unchanged. However, no significant correlation was observed between MDR1 expression and the patients' survival or treatment response. CONCLUSIONS: Modulation of MDR1 gene expression in PMNs after lung cancer treatment with paclitaxel and carboplatin cannot be used as a prognosis marker in these patients.
Cancer Chemotherapy and Pharmacology 11/2012; · 2.83 Impact Factor
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María Berdasco,
Consolación Melguizo, Jose Prados,
Antonio Gómez,
Miguel Alaminos,
Miguel A Pujana,
Miguel Lopez,
Fernando Setien,
Raul Ortiz,
Inma Zafra,
Antonia Aranega,
Manel Esteller
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ABSTRACT: Adult stem cells have an enormous potential for clinical use in regenerative medicine that avoids many of the drawbacks characteristic of embryonic stem cells and induced pluripotent stem cells. In this context, easily obtainable human adipose-derived stem cells offer an interesting option for future strategies in regenerative medicine. However, little is known about their repertoire of differentiation capacities, how closely they resemble the target primary tissues, and the potential safety issues associated with their use. DNA methylation is one of the most widely recognized epigenetic factors involved in cellular identity, prompting us to consider how the analyses of 27,578 CpG sites in the genome of these cells under different conditions reflect their different natural history. We show that human adipose-derived stem cells generate myogenic and osteogenic lineages that share much of the DNA methylation landscape characteristic of primary myocytes and osteocytes. Most important, adult stem cells and in vitro-generated myocytes and osteocytes display a significantly different DNA methylome from that observed in transformed cells from these tissue types, such as rhabdomyosarcoma and osteosarcoma. These results suggest that the plasticity of the DNA methylation patterns plays an important role in lineage commitment of adult stem cells and that it could be used for clinical purposes as a biomarker of efficient and safely differentiated cells.
American Journal Of Pathology 09/2012; · 4.89 Impact Factor
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ABSTRACT: The potential use of gene therapy to improve the response of patients with advanced cancer is being intensively analyzed. We evaluated the cytotoxic impact of the gef gene, a suicide gene, which has a demonstrated antiproliferative activity in tumor cells, in colon carcinoma cells in order to improve the antitumour effect of chemotherapeutic drugs used as first line treatment in the management of advanced colon cancer. We found that the gef gene induced a marked decrease in cell viability (50% in 24h) in T-84 cells through cell death by apoptosis. Interestingly, when gef gene expression was combined with drugs of choice in the clinical treatment of colon cancer (5-fluorouracil, oxaliplatin and irinotecan), a strong synergistic effect was observed with approximately a 15-20% enhancement of the antiproliferative effect. Our data demonstrate, for the first time, that gef gene expression induces significant growth arrest in colon cancer cells and that it is able to enhance the effect of some cytotoxic drugs compared with a single therapeutic approach. These results indicate the potential therapeutic value of the gef gene in colon cancer combination therapy.
Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 06/2012; 66(7):563-7. · 2.24 Impact Factor
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ABSTRACT: Cancer is the second leading cause of death in the Western world. The limited successes of available treatments for cancer mean that new strategies need to be developed. The possibility of modifying the cancer cell with the introduction of genetic material opens the way to a new approach based on gene therapy. There are still many technical difficulties to be overcome, but recent advances in the molecular and cellular biology of gene transfer have made it likely that gene therapy will soon start to play an increasing role in clinical practice, particularly in the treatment of cancer. Gene therapy will probably be the therapeutic option in cases in which conventional treatments such as surgery, radiotherapy and chemotherapy have failed. The development of modified vectors, and an improved understanding of interactions between the vector and the human host, are generating inventions that are being protected by patents due to the considerable interest of industry for their possible commercialization. We review the latest strategies, patented and/or under clinical trial, in cancer gene therapy. These include patents that cover the use of modified vectors to increase the security and specificity, recombining adenovirus that leads to loss or gain of gene function, activation of the patient's own immune cells to eliminate cancer cells by expression of molecules that enhance immune responses, silencing genes related to the development of drug resistance in patients, inhibition of angiogenesis of solid tumors by targeting the tumor vasculature, and the development of enzymes that destroy viral or cancerous genetic material.
Recent patents on anti-cancer drug discovery. 02/2012; 7(3):297-312.
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ABSTRACT: INTRODUCTION: The fluorinated analog of uracil 5-FU is an antimetabolite, active against a wide range of solid tumors. The main mechanism of action consists in interfering with DNA synthesis and mRNA translation. However, patients treated with 5-FU display several side effects, a result of its nonspecific cytotoxicity for tumor cells. Numerous modifications of the 5-FU structure have been performed in order to overcome these disadvantages. AREAS COVERED: In this review, the metabolic pathways, pharmacokinetics and clinical pharmacology of 5-FU are briefly introduced. Moreover, several derivatives developed and patented, including oral 5-FU prodrugs and combinations with other active compounds, are presented. Finally, new innovative methods for administration and vehiculization of 5-FU and its derivatives are described. EXPERT OPINION: The search for less toxic 5-FU derivatives, which diminish or circumvent some of its disadvantages, has allowed the development of selective antitumor prodrugs and novel methods for tissue-specific drug delivery. Although some of these oral prodrugs are being used clinically, either alone or in combination therapy with other anticancer agents, it seems that the potential of personalized medicine, including pharmacogenomics and targeted therapy with novel 5-FU derivatives, will improve the management and clinical responses of patients treated with 5-FU-based therapy.
Expert Opinion on Therapeutic Patents 02/2012; 22(2):107-23. · 3.57 Impact Factor
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ABSTRACT: Carboplatin-paclitaxel is a reference regimen in the treatment of locally advanced or disseminated non-small cell lung cancer (NSCLC). This paper discusses the multidrug resistance developed with this drug combination, which is one of the major obstacles to successful treatment. In order to understand and overcome the drug resistance pattern of NSCLC after carboplatin plus paclitaxel exposure, levels of mRNA expression of multidrug resistance 1 (MDR1) and multidrug resistance-associated protein 3 (MRP3) were investigated in primary NSCLC cell lines (A-549 and A-427) and a metastasis-derived NSCLC cell line (NODO). Our results showed that exposure of the three NSCLC lines to plasma concentrations of paclitaxel (5 μM) produced an increase in MDR1 expression, while MRP3 showed no alteration in expression. By contrast, the same cells exposed to carboplatin plasma concentrations (30 μM) showed overexpression of MRP3. In these cells, MDR1 showed no expression changes. Interestingly, the combination of both paclitaxel and carboplatin caused increased expression of the MDR1 drug resistance gene rather than the individual treatments. These results suggest that carboplatin and paclitaxel may induce drug resistance mediated by MDR1 and MRP3, which may be enhanced by the simultaneous use of both drugs.
International Journal of Molecular Sciences 01/2012; 13(12):16624-35. · 2.60 Impact Factor
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ABSTRACT: Classical cytotoxic treatment of rhabdomyosarcoma (RMS) is often accompanied by significant morbidity and poor response. This cytotoxic therapy may induce a multidrug resistance (MDR) phenotype in RMS which is associated with decreased effectiveness of chemotherapy. The majority of MDR molecules belong to a family of ABC (ATP binding cassette) transporters. Studies of drug resistance in RMS suggest that there are various mechanisms acting simultaneously, which might explain the low percentage of long-term survival in this malignancy. Moreover, although cells exposed to cytotoxic agents increase expression of muscle differentiation markers indicating myogenic differentiation, multidrug resistance may be a major obstacle in differentiation therapy for RMS. This review briefly discusses the current knowledge of resistance in RMS and emphasizes the importance of understanding the different aspects of MDR status in these patients.
Oncology Reports 06/2011; 26(4):755-61. · 1.84 Impact Factor
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ABSTRACT: The limited ability of conventional therapies to achieve the long-term survival of metastatic lung and colon cancer patients suggests the need for new treatment options. In this respect, genes encoding cytotoxic proteins have been proposed as a new strategy to enhance the activity of drugs, and combined therapies involving such genes and classical antitumoral drugs have been studied intensively. The E gene from phiX174 encodes a membrane protein with a toxic domain that leads to a decrease in tumour cell growth rates. Therefore, in order to improve the anti-tumour effects of currently used chemotherapeutic drugs on cancer cells, we investigated the association of the E suicide gene with these antineoplastic drugs. The E gene has antitumoral effects in both lung and colon cancer cells. In addition, expression of this gene induces ultrastructural changes in lung cancer transfected cells (A-549), although the significance of these changes remains unknown. The effect of combined therapy (gene and cytotoxic therapy) enhances the inhibition of tumour cell proliferation in comparison to single treatments. Indeed, our in vitro results indicate that an experimental therapeutic strategy based on this combination of E gene therapy and cytotoxic drugs may result in a new treatment strategy for patients with advanced lung and colon cancer.
Bioengineered bugs 05/2011; 2(3):163-7.
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ABSTRACT: The low effectiveness of conventional therapies to achieve the long-term survival of metastatic breast cancer patients calls for the development of novel options. Genes encoding cytotoxic proteins have been proposed as a new strategy to enhance the antiproliferative activity of drugs. Combined therapy using these genes and classical antitumoral drugs are under intensive study. The E gene from ϕX174 encodes a membrane protein with a toxic domain that leads to a decrease in the tumour cell growth rate. With the aim of improving the anti-tumour effect on breast cancer cells of the currently used chemotherapeutic drugs (Paclitaxel, Docetaxel and Doxorubicin), we investigated the association of E suicide gene with these drugs. The effect of the combined therapy (gene therapy and cytotoxic) was determined by treating transfected MCF-7 cells and multicellular tumour spheroids (MTS) with drugs gradient concentrations. Our results showed that E gene has a direct oncolytic effect inducing a significant decrease in the proliferation rate of the MCF-7 cells. The E gene antitumoral activity was mediated by the induction of apoptosis (mitochondrial pathway). In addition, a significant enhancement of proliferation inhibition was observed when E gene transfection was associated with cytotoxic drugs in comparison to single treatments. The use of the combined therapy E gene-Doxorubicin obtained the greatest effect on the MCF-7 growth arrest. This therapeutic association also induced a significant enhancement of the MTS volume growth inhibition. Anti-tumour activity of the chemotherapeutic drugs classically used in the treatment of breast cancer was enhanced by E gene. Our in vitro results indicate that experimental therapeutic strategy based in the combined therapy E gene and cytotoxic drugs may be of potential therapeutic value as a new strategy for patients with advanced breast cancer.
Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 04/2011; 65(4):260-70. · 2.24 Impact Factor
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ABSTRACT: Nanotechnology, along with related concepts such as nanomaterials, nanostructures and nanoparticles, has become a priority area for scientific research and technological development. Nanotechnology, i.e., the creation and utilization of materials and devices at nanometer scale, already has multiple applications in electronics and other fields. However, the greatest expectations are for its application in biotechnology and health, with the direct impact these could have on the quality of health in future societies. The emerging discipline of nanomedicine brings nanotechnology and medicine together in order to develop novel therapies and improve existing treatments. In nanomedicine, atoms and molecules are manipulated to produce nanostructures of the same size as biomolecules for interaction with human cells. This procedure offers a range of new solutions for diagnoses and "smart" treatments by stimulating the body's own repair mechanisms. It will enhance the early diagnosis and treatment of diseases such as cancer, diabetes, Alzheimer's, Parkinson's and cardiovascular diseases. Preventive medicine may then become a reality.
International Journal of Molecular Sciences 01/2011; 12(5):3303-21. · 2.60 Impact Factor
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Houria Boulaiz,
Pablo J Alvarez, Jose Prados,
Juan Marchal,
Consolación Melguizo,
Esmeralda Carrillo,
Macarena Peran,
Fernando Rodríguez,
Alberto Ramírez,
Raúl Ortíz,
Antonia Aránega
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ABSTRACT: Breast cancer research has developed rapidly in the past few decades, leading to longer survival times for patients and opening up the possibility of developing curative treatments for advanced breast cancer. Our increasing knowledge of the biological pathways associated with the progression and development of breast cancer, alongside the failure of conventional treatments, has prompted us to explore gene therapy as an alternative therapeutic strategy. We previously reported that gef gene from E. coli has shown considerable cytotoxic effects in breast cancer cells. However, its action mechanism has not been elucidated. Indirect immunofluorescence technique using flow cytometry and immunocytochemical analysis were used to detect breast cancer markers: estrogen (ER) and progesterone (PR) hormonal receptors, human epidermal growth factor receptor-2 proto-oncogene (c-erbB-2), ki-67 antigen and p53 protein. gef gene induces an increase in ER and PR expressions and a decrease in ki-67 and c-erbB-2 gene expressions, indicating a better prognosis and response to treatment and a longer disease-free interval and survival. It also increased p53 expression, suggesting that gef-induced apoptosis is regulated by a p53-mediated signaling pathway. These findings support the hypothesis that the gef gene offers a new approach to gene therapy in breast cancer.
International Journal of Molecular Sciences 01/2011; 12(11):7445-58. · 2.60 Impact Factor
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ABSTRACT: The E gene from ΦX174 encodes a membrane protein with a toxic domain that leads to a decrease in the tumour cell growth rate. With the aim of improving the antitumour effect on lung and colon cancer cells of the currently used chemotherapeutic drugs such as gemcitabine, carboplatin and paclitaxel, and 5-fluorouracil (5FU) plus folinic acid (FA) with irinotecan or oxaliplatine, we investigated a new combined therapy using these drugs associated to the transfection of E gene. Our results showed that E gene was able to decrease proliferation rate in A-549 and T-84 cells by inducing apoptotic the mitochondrial pathway. Significantly greater inhibition of proliferation was obtained using drugs in combination with E gene in comparison to single-agent treatments or controls. E gene combined with paclitaxel had the greatest effect on A-549 cells and combined with 5FU/FA/oxaliplatin on T-84 cells. Antitumour mechanisms of the chemotherapeutic drugs were enhanced by E gene, which itself has direct oncolytic effects inducing A-549 and T-84 apoptosis. Our in vitro results indicate that the combined therapy of E gene and cytotoxic drugs may be of potential therapeutic value as a new strategy for patients with advanced lung and colon cancer.
International Journal of Oncology 12/2010; 37(6):1503-14. · 2.40 Impact Factor
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ABSTRACT: The potential use of combined therapy is under intensive study including the association between classical cytotoxic and genes encoding toxic proteins which enhanced the antitumour activity. The main aim of this work was to evaluate whether the gef gene, a suicide gene which has a demonstrated antiproliferative activity in tumour cells, improved the antitumour effect of chemotherapeutic drugs used as first-line treatment in the management of advanced breast cancer.
MCF-7 human breast cancer cells were transfected with gef gene using pcDNA3.1-TOPO expression vector. To determine the effect of the combined therapy, MCF-7 transfected and non-transfected cells were exposed to paclitaxel, docetaxel and doxorubicin at different concentrations. The growth-inhibitory effect of gef gene and/or drugs was assessed by MTT assay. Apoptosis modulation was determined by flow cytometric analysis, DNA fragmentation and morphological analysis. Multicellular tumour spheroids (MTS) from MCF-7 cells were used to confirm effectiveness of combined therapy (gef gene and drug).
Our results demonstrate that combined therapy gef gene/drugs (paclitaxel, docetaxel or doxurubicin) caused a decrease in cell viability. However, only the gef-doxorubicin (10 microM) combination induced a greater enhancement in the antitumour activity in MCF-7 cells. Most importantly, this combined strategy resulted in a significant synergistic effect, thus allowing lower doses of the drug to be used to achieve the same therapeutic effect. These results were confirmed using MTS in which volume decrease with combined therapy was greater than obtained using the gene therapy or chemotherapy alone, or the sum of both therapies.
The cytotoxic effect of gef gene in breast cancer cells enhances the chemotherapeutic effect of doxorubicin. This therapeutic approach has the potential to overcome some of the major limitations of conventional chemotherapy, and may therefore constitute a promising strategy for future applications in breast cancer therapy.
Cancer Chemotherapy and Pharmacology 09/2009; 66(1):69-78. · 2.83 Impact Factor
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ABSTRACT: Novel treatment modalities, including gene therapy, are needed for patients with advanced melanoma. We evaluated whether the gef gene, a suicide gene from Escherichia coli, had a significant cytotoxic impact on melanoma in vivo. First, we used a non-viral gene delivery approach (pcDNA3.1/gef) to study the inhibition of melanoma cells (B16-F10) proliferation in vitro. Secondly, we used direct intra-tumoral injection of pcDNA3.1/gef complexed with jetPEI to deliver gef cDNA to rapidly growing murine melanomas. We demonstrated that gef gene not only has an antiproliferative effect on B16-F10 cells in vitro, but also induces an important decrease in melanoma tumor volume (77.7% in 8 days) in vivo. Interestingly, after gef gene treatment, melanoma showed apoptosis activation associated with the mitochondrial pathway, suggesting that the induction of this death mechanism may be an effective strategy for its treatment. Our in vivo results indicate that gef gene might become a suitable therapeutic strategy for patients with advanced melanoma.
Experimental Dermatology 08/2009; 19(4):363-71. · 3.54 Impact Factor
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Raúl Ortiz, Jose Prados,
Consolacion Melguizo,
Ana R Rama,
Ana Segura,
Fernando Rodríguez-Serrano,
Houria Boulaiz,
Fidel Hita,
Antonio Martinez-Amat,
Roberto Madeddu,
Juan L Ramos,
Antonia Aranega
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ABSTRACT: Novel treatment modalities, including gene therapy, are needed for patients with advanced melanoma. The E gene from the phage varphiX174 encodes a 91-aa protein which lyses Escherichia coli by formation of a transmembrane tunnel structure. To evaluate whether this E gene has a cytotoxic impact on melanoma cells in vitro and in vivo, and could therefore be used as a new therapeutic strategy for this tumor type, we selected the B16-F10 murine melanoma cell line as a model. We used a nonviral gene delivery approach (pcDNA3.1/E plasmid) to study the inhibition of melanoma cells' proliferation in vitro and direct intratumoral injection of pcDNA3.1/E complexed with jetPEI to deliver E cDNA to rapidly growing murine melanomas, and found that the E gene has both a strong antiproliferative effect in B16-F10 cells in vitro and induces an efficient decrease in melanoma tumor volume in vivo (90% in 15 days). Interestingly, the GFP-E fusion protein expressed in melanoma cells was located in the mitochondria. In vitro and in vivo analysis demonstrated significant functional and morphological mitochondrial alterations accompanied by a significant increase of cytochrome c and active caspase-3 and -9 in transfected cells, which suggests that tumoral cell death is mediated by the mitochondrial apoptotic pathway. These results show that E gene expression in melanoma cells has an extraordinary antitumor effect, which means it may be a new candidate for an effective strategy for melanoma treatment.
Journal of Molecular Medicine 08/2009; 87(9):899-911. · 4.67 Impact Factor
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ABSTRACT: The low efficiency of conventional therapies in achieving long-term survival of lung cancer patients calls for development of novel options. The potential use of combined gene therapy is under intensive study. One approach uses the expression of genes encoding cytotoxic proteins that affect cellular viability. The gef gene from E. coli, identified as a member of a gene family encoding homologous cell-killing functions, encodes for a membrane protein with a toxic domain which leads to a decrease in the rate of tumour cell growth. To improve the antitumoral effect of the paclitaxel in lung cancer cells, we investigated a combined suicide gene therapy using this drug and gef gene in vitro, using A-549 lung cancer cells in culture and forming multicellular tumour spheroids (MTS). Our results showed that gef expression in A-549 cells led to an ultrastructural changes, including dilated mitochondria with clear matrices and disrupted cristae and cell surface alterations such as reduction in length and number of microvilli and cytoplasmic membrane evaginations. The use of paclitaxel in A-549 lung cancer cells transfected with gef gene enhanced the chemotherapeutic effect of this drug. Volume analyses showed an 87.4% decrease in the A-549 MTS growth after 96 h in comparison with control MTS. This inhibition was greater than that obtained using the gene therapy or chemotherapy alone. In conclusion, gef gene has a cytotoxic effect in lung cancer cells and enhances cell growth inhibition when used with paclitaxel. These results indicate that this combined therapy may be of potential therapeutic value in lung cancer.
International Journal of Oncology 08/2008; 33(1):121-7. · 2.40 Impact Factor
