RESUMEN
Diabetes mellitus (DM) is a complex disease that affects many systems. The most important cells of the immune system are lymphomononuclear (LMN) cells. Here, we aimed to evaluate the energy metabolism of LMN cells in patients with diabetes and impaired glucose tolerance. We measured LMN cell energy metabolism in patients with type 2 diabetes mellitus, impaired glucose tolerance (IGT) and healthy subjects. Cells were freshly isolated from peripheral blood and the subgroups were determined by flow cytometric method. Lactate production and glycogen utilization were significantly increased in the LMN cells of patients with type 2 DM and IGT when compared with healthy volunteers. No statistical difference was observed between the patients with type 2 DM and IGT. There was a significant correlation between fasting plasma glucose and lactate production in LMN cells. LMN cells changed their energy pathway in a diabetic state and preferred anaerobic glycolysis. Prediabetic range also affected energy metabolism in LMN cells. This abnormal energy production might cause dysfunction in LMN cells and the immune system in diabetic and prediabetic patients. In conclusion, we concluded that impaired glucose metabolism could change energy metabolism.
Asunto(s)
Diabetes Mellitus Tipo 2/metabolismo , Metabolismo Energético , Intolerancia a la Glucosa/metabolismo , Leucocitos Mononucleares/metabolismo , Anciano , Glucemia/análisis , Células Cultivadas , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/inmunología , Femenino , Intolerancia a la Glucosa/sangre , Intolerancia a la Glucosa/inmunología , Glucogenólisis , Glucólisis , Humanos , Hiperglucemia/inmunología , Hiperglucemia/metabolismo , Ácido Láctico/metabolismo , Masculino , Persona de Mediana Edad , Estado Prediabético/sangre , Estado Prediabético/inmunología , Estado Prediabético/metabolismoAsunto(s)
Factores de Coagulación Sanguínea/metabolismo , Terapia Genética/métodos , Trastornos Hemorrágicos/terapia , Biología Molecular/métodos , Inductores de la Angiogénesis/metabolismo , Factores de Coagulación Sanguínea/uso terapéutico , Progresión de la Enfermedad , Terapia Genética/tendencias , Trastornos Hemorrágicos/genética , Hepatocitos/química , Hepatocitos/trasplante , Humanos , Hígado/química , Células Madre Mesenquimatosas/química , Biología Molecular/tendencias , ARN Mensajero/metabolismo , Proteínas Recombinantes/uso terapéuticoRESUMEN
In B-cell chronic lymphocytic leukaemia (B-CLL), cutaneous infiltration is far less common than in T-cell CLL. However, the effect of fludarabine on cutaneous infiltration in patients with B-CLL is uncertain. We describe a 63-year-old man with B-CLL presenting with cutaneous lesions, who was treated successfully with oral fludarabine. Skin biopsy of one of these lesions revealed diffuse infiltration of uniform lymphocytes. Using PCR analysis, the same immunoglobulin heavy-chain gene rearrangement was found in lymphocytes in all samples (peripheral blood, bone marrow and skin lesion). The patient received four courses of oral fludarabine. Simultaneously with the normalization of the peripheral blood lymphocytosis, the patient became free of the cutaneous infiltration of CLL after four courses of oral fludarabine. Skin lesions had not recurred by the 12-month follow-up examination. To our knowledge, this is the first case of B-CLL leukaemia cutis treated with oral fludarabine. The introduction of fludarabine may contribute to a better outlook for these patients in the future.
Asunto(s)
Antineoplásicos/uso terapéutico , Leucemia Linfocítica Crónica de Células B/tratamiento farmacológico , Infiltración Leucémica/patología , Piel/patología , Vidarabina/análogos & derivados , Administración Oral , Anciano , Humanos , Leucemia Linfocítica Crónica de Células B/patología , Infiltración Leucémica/tratamiento farmacológico , Masculino , Resultado del Tratamiento , Vidarabina/uso terapéuticoRESUMEN
AIM: To investigate whether granulocyte-macrophage colony-stimulating factor (GM-CSF) with or without thalidomide can induce apoptosis and differentiation of HL-60 acute promyelocytic leukemia cell line in vitro. METHODS: Effect of GM-CSF and thalidomide on proliferation of HL-60 cells was evaluated by MTT assay, cell cycle analysis was performed by propidium iodide staining approach and flow cytometry, and apoptosis rate was analyzed using FITC-conjugated annexin-V and FACScan flow cytometry. RESULTS: The study revealed that thalidomide alone at high concentrations inhibited HL-60 cell growth and induced apoptosis. Three days treatment of low-dose thalidomide in combination with GM-CSF induced marked terminal differentiation of HL-60 cells, as it was assessed by increased expression of differentiation antigens on cell surface. CONCLUSION: Treatment of HL-60 cells by low concentration of thalidomide combined with GM-CSF induced terminal differentiation of HL60 cells in vitro, which may be advantageous for the elaboration of novel therapeutic regimens in patients with differentiation-inducible leukemias.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Leucemia Promielocítica Aguda/patología , Talidomida/farmacología , Antígenos de Diferenciación/análisis , Antígenos de Diferenciación/metabolismo , Apoptosis , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células HL-60 , Humanos , Leucemia Promielocítica Aguda/metabolismoRESUMEN
Prediabetes has been associated with an increased risk of cardiovascular disease and mortality. Soluble P-selectin (sP-selectin) is an index of platelet activation and also a risk factor for future vascular events. sP-selectin levels were investigated in prediabetic subjects who had no confounding factors such as hypertension, obesity or dyslipidaemia. sP-selectin, hsCRP levels and HOMA-IR indexes were measured in 40 prediabetic subjects (n = 24 for IFG and n = 16 for IGT) and age-, sex- and BMI-matched 40 healthy controls. sP-selectin levels in prediabetic subjects were not significantly different compared with those in controls (p = 0.12). Prediabetic group had similar hsCRP (p = 0.29), higher HOMA-IR indexes (p < 0.001) and lower HDL cholesterol levels (p = 0.001) when compared with healthy controls. The power of the study was 0.93 for sP-selectin, 0.7 for hsCRP and 1.0 for HOMA. Our data suggest that sP-selectin may not contribute to the prothrombotic state as well as the accelerated atherogenesis associated with prediabetes.
Asunto(s)
Selectina-P/sangre , Estado Prediabético/sangre , Proteína C-Reactiva/metabolismo , Estudios de Casos y Controles , Femenino , Intolerancia a la Glucosa/sangre , Humanos , Masculino , Persona de Mediana Edad , Factores de RiesgoRESUMEN
AIM: The goals of the study were to reveal the involvement of P-glycoprotein (P-gp), the product of multidrug resistance 1 gene (MDR1) in cellular resistance to vincristine (VCR) and investigate cross-resistance against cytosine arabinoside (Ara-C) in HL60 and HL60/VCR cell lines. METHODS: HL60 cells (human acute myeloid leukemia cell line) were cultured on medium with 1-50 nM of VCR for 4-6 weeks, and VCR resistant cells (HL60/VCR) were selected. The viability of cells was assessed by MTT assay and the expression of MDR1 gene was detected by RT-PCR. RESULTS: No expression of MDR1 gene was revealed in HL60 cells, but MDR1 started to be expressed after incubation of cells with 2 nM of VCR and its expression level elevates with increase of agent concentration. MTT test has shown that HL-60/VCR cells were 75-fold more resistant to VCR and 42-fold higher resistant to cytosine arabinoside (Ara-C) compared to sensitive HL60 cells. CONCLUSION: Aquired resistance to VCR and cross-resistance to Ara-C correlates with MDR1 gene expression in vitro.
Asunto(s)
Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Antineoplásicos/farmacología , Citarabina/farmacología , Leucemia Mieloide/genética , Vincristina/farmacología , Enfermedad Aguda , Resistencia a Antineoplásicos/genética , Expresión Génica , Humanos , Selección Genética , Células Tumorales CultivadasRESUMEN
Little information is available on the immunological basis for side-effects of dental materials. The objective of this study is to evaluate effects of pure metals, dental alloys and ceramic on cell viability and interleukin-1 beta (IL-1beta) release in three-dimensional human gingival fibroblast cultures as an indicator of their biological performance in gingival tissues. The gingival fibroblast cultures were exposed to test specimens fabricated from nickel, iron, molybdenum, copper, indium, gold, Ni-Cr-Mo alloy (Remanium CS), Au-Pt-In alloy (Pontostar) and a dental ceramic (In-ceram). Cell viability was determined by the MTT method 24 and 48 h after exposure. Assays for IL-1beta were carried out by ELISA. Statistical analysis was performed applying the non-parametric Mann-Whitney pairwise test. Dental ceramic and gold did not influence cell viability after 24 and 48 h. Cell viability was determined after 24 and 48 h to nickel (79-77%), iron (92-90%), molybdenum (86-83%), copper (48-36%), indium (90-90%), Remanium CS (83-80%), Pontostar (94-91%) compared with control cultures. Dental ceramic, Pontostar and gold had no significant influence on IL-1beta secretion. The highest amounts of IL-1beta (10-fold) levels were determined in cell cultures exposed to copper. Indium, molybdenum and iron induced twofold IL-1beta levels compared with untreated control cultures. These results support that some metals may alter immune responses and thereby contribute to a variety of dental pathological conditions and three-dimensional cell culture models for gingival fibroblasts appear to be suitable for in vitro studies.
Asunto(s)
Cerámica/farmacología , Aleaciones Dentales/farmacología , Fibroblastos/efectos de los fármacos , Encía/efectos de los fármacos , Interleucina-1/metabolismo , Metales/farmacología , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Cobre/farmacología , Fibroblastos/inmunología , Encía/inmunología , Oro/farmacología , Humanos , Indio/farmacología , Hierro/farmacología , Molibdeno/farmacología , Níquel/farmacología , Estadísticas no ParamétricasAsunto(s)
Trasplante de Médula Ósea/fisiología , Factor Estimulante de Colonias de Granulocitos/uso terapéutico , Neutrófilos/fisiología , Neutrófilos/trasplante , Trasplante de Células Madre , Adolescente , Adulto , Transfusión Sanguínea , Niño , Filgrastim , Humanos , Terapia de Inmunosupresión/métodos , Inmunosupresores/uso terapéutico , Leucemia/terapia , Recuento de Leucocitos , Linfoma no Hodgkin/terapia , Persona de Mediana Edad , Neutrófilos/efectos de los fármacos , Proteínas Recombinantes , Estudios Retrospectivos , Factores de Tiempo , Trasplante Homólogo , Resultado del Tratamiento , Irradiación Corporal TotalRESUMEN
OBJECTIVE: The aim of this study was to determine whether a relationship exists between gastric and oral Helicobacter pylori and oral hygiene in patients with vitamin B12 deficiency. STUDY DESIGN: One hundred eight patients with vitamin B12 deficiency who were H pylori -positive in their gastric mucosa were enrolled in the study. These patients were divided into 3 groups determined by Oral Hygiene Index (OHI) scores of good, fair, or poor. H pylori was detected in the dental plaque with camphylobacter-like organism test gels. All patients were treated with a combination regimen to eradicate H pylori. RESULTS: H pylori positivity in dental plaque was correlated with OHI scores; the positivity was 28.5%, 90.2%, or 100% in patients with good, fair, or poor OHI scores, respectively. The eradication of H pylori was associated with recovery from anemia and increased serum vitamin B12 level (P <.0001 and P <.0001). The patients with poor OHI scores had the most frequent gastric recurrence of H pylori (58.3%) compared with those with fair OHI scores (41.2%) and good OHI scores (4.8%). CONCLUSIONS: H pylori seems to be an etiologic factor in vitamin B12 deficiency, since anemia was cured and the level of vitamin B12 in the serum increased as a result of its eradication. However, eradication of H pylori from gastric mucosa alone is not enough to prevent gastric recurrence of the bacteria. Proper oral hygiene must be established to eliminate H pylori in dental plaque. Therefore, we suggest that control of H pylori in dental plaque is necessary to control recurrence of H pylori.
Asunto(s)
Anemia Perniciosa/microbiología , Placa Dental/microbiología , Mucosa Gástrica/microbiología , Helicobacter pylori/fisiología , Higiene Bucal , Deficiencia de Vitamina B 12/microbiología , Adulto , Anciano , Anciano de 80 o más Años , Amoxicilina/uso terapéutico , Anemia Perniciosa/terapia , Antibacterianos/uso terapéutico , Antiulcerosos/uso terapéutico , Campylobacter/clasificación , Distribución de Chi-Cuadrado , Claritromicina/uso terapéutico , Cálculos Dentales/clasificación , Femenino , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/tratamiento farmacológico , Helicobacter pylori/efectos de los fármacos , Humanos , Masculino , Persona de Mediana Edad , Omeprazol/uso terapéutico , Índice de Higiene Oral , Penicilinas/uso terapéutico , Recurrencia , Estadística como Asunto , Gastropatías/tratamiento farmacológico , Gastropatías/microbiología , Vitamina B 12/sangre , Deficiencia de Vitamina B 12/terapiaRESUMEN
Several reports have noted pancytopenia associated with Human parvovirus B19 (PVB19) or Ebstein-Barr virus (EBV) infections in patients who have no history of immunodeficiency. To our knowledge, we report the first case of severe aplastic anemia associated with both EBV and PVB19 infections in a previously healthy 22-year-old man. He was admitted to our hematology service due to anemia and thrombocytopenia. He had no symptoms or signs of infections of these viruses. His bone marrow biopsy revealed a hypocellular marrow. Specific IgM and IgG antibodies to EBV and PVB19 were elevated. EBV and PVB19 virus genomes were detected by PCR in the bone marrow nucleated cells and the peripheral blood lymphocytes. Two months after treatment with prednisone, acyclovir, and intravenous immune globulin (IVIg), the genomes of both these viruses disappeared. However, his transfusion requirement for platelet suspensions and packed red blood cells persisted. The patient underwent allogeneic bone marrow transplant (allo-BMT) and has had an enduring complete hematological response for 8 months.
Asunto(s)
Anemia Aplásica/virología , Trasplante de Médula Ósea , Infecciones por Virus de Epstein-Barr/complicaciones , Infecciones por Parvoviridae/complicaciones , Adulto , Anemia Aplásica/etiología , Anemia Aplásica/terapia , Médula Ósea/patología , Médula Ósea/virología , Infecciones por Virus de Epstein-Barr/tratamiento farmacológico , Humanos , Masculino , Infecciones por Parvoviridae/tratamiento farmacológico , Transfusión de Plaquetas , Trasplante Homólogo , Resultado del TratamientoRESUMEN
Erythema annulare centrifugum is a figurate erythema that has been associated with many different entities. A case of erythema annulare centrifugum related to non-Hodgkin's lymphoma in a 38-year-old woman is described in this case report. Response of the lymphoma to a combination chemotherapy was accompanied by disappearance of skin lesions. When therapy was discontinued, both disorders recurred, and both responded to reinstitution of a different chemotherapy regimen. To our knowledge, this case is the first reported association of erythema annulare centrifugum and non-Hodgkin's lymphoma in the medical literature.
Asunto(s)
Eritema/etiología , Linfoma de Células B Grandes Difuso/complicaciones , Adulto , Eritema/inmunología , Femenino , Humanos , Antígeno Ki-1 , Linfoma de Células B Grandes Difuso/inmunologíaRESUMEN
Abelcet is composed of large particles which could be misinterpreted as blood cells on measurements with blood cell counters. The direct measurement of drug suspensions on blood cell counters has been performed in an in vitro study. In an ex vivo study, the haematological parameters were compared before and during Abelcet infusions. A significant interference effect was observed in platelet counts, together with minimal differences in the WBC, RBC, and Hb parameters in the in vitro study. In the ex vivo study, there were statistically significant deviations only in RBC counts and in haemoglobin (Hb) level, while there was no difference in the other parameters. It is been reported that the drug accumulates very rapidly in the reticuloendothelial system and circulates minimally in the plasma. That is why there is a significant deviation in the direct counting of platelets, while the platelet counts taken from the patients do not differ statistically.
Asunto(s)
Anfotericina B/farmacología , Errores Diagnósticos/instrumentación , Pruebas Hematológicas/normas , Fosfatidilcolinas/farmacología , Fosfatidilgliceroles/farmacología , Adulto , Anfotericina B/administración & dosificación , Aspergilosis Broncopulmonar Alérgica/sangre , Aspergilosis Broncopulmonar Alérgica/tratamiento farmacológico , Recuento de Células Sanguíneas/instrumentación , Recuento de Células Sanguíneas/normas , Linfoma de Burkitt/sangre , Linfoma de Burkitt/tratamiento farmacológico , Linfoma de Burkitt/microbiología , Errores Diagnósticos/prevención & control , Combinación de Medicamentos , Recuento de Eritrocitos/instrumentación , Recuento de Eritrocitos/normas , Femenino , Pruebas Hematológicas/instrumentación , Hemoglobinas/análisis , Humanos , Masculino , Fosfatidilcolinas/administración & dosificación , Fosfatidilgliceroles/administración & dosificación , Recuento de Plaquetas/instrumentación , Recuento de Plaquetas/normas , Reproducibilidad de los ResultadosRESUMEN
Severe 5-fluorouracil (5-FU) toxicity has been reported among patients lacking dihydropyrimidine dehydrogenase (DPD) enzymatic activity. DPD is the principal enzyme involved in the degradation of 5-FU to 5'-6'-dihydrofluorouracil, which is further metabolized to fluoro-beta-alanine. We demonstrate here that overexpression of human DPD confers resistance to 5-FU in NIH3T3 cells, mouse bone marrow cells, and in human CD34+-enriched hematopoietic progenitor cells. An SFG-based dicistronic retroviral vector containing human DPD cDNA, an internal ribosomal entry site (IRES), and the neomycin phosphotransferase (Neo) gene was constructed (SFG-DPD-IRES-Neo). Transduced NIH3T3 cells demonstrated a 2-fold (ED50) increase in resistance to a 4-hour exposure of 5-FU in comparison to nontransduced cells. Expression of DPD was confirmed by Northern and Western blot analyses, and DPD enzyme activity was detectable only in transduced cells. Infection of mouse bone marrow cells with this retroviral construct resulted in an increased number of 5-FU-resistant CFU-GM colonies, compared to mock-transduced bone marrow in both 4-hour and 12- to 14-day exposures. Infection of human CD34+-enriched cells with this construct and incubation with 5-FU (10(-6) M) for 14 days also resulted in an increased number of 5-FU-resistant colonies. Retroviral transduction of human hematopoietic progenitor cells with a cDNA-expressing human DPD conferred resistance to 5-FU in NIH3T3 cells, mouse bone marrow cells, and human CD34+-enriched cells. These results encourage the use of this gene as a method to protect patients from 5-FU myelotoxicity.
Asunto(s)
Resistencia a Medicamentos/genética , Células Madre Hematopoyéticas/fisiología , Oxidorreductasas/genética , Células 3T3 , Animales , Antimetabolitos Antineoplásicos/toxicidad , ADN Complementario/biosíntesis , ADN Complementario/genética , Dihidrouracilo Deshidrogenasa (NADP) , Fluorouracilo/toxicidad , Vectores Genéticos , Células Madre Hematopoyéticas/efectos de los fármacos , Humanos , Inmunosupresores/toxicidad , Ratones , Oxidorreductasas/biosíntesis , Retroviridae , TransfecciónRESUMEN
An 80-year old man was diagnosed as having immune thrombocytopenic purpura based on epistaxis, purpura and by the platelet count 8 x 10(9)/l. Prednisolone and gamma globulin were administered and the platelet count had been kept around 50 x 10(9)/l during his follow up. Two years from the onset of immune thrombocytopenic purpura he was admitted because of leukocytosis (79 x 10(9)/l with 79% monocytes), anemia and thrombocytopenia. Hypercellular bone marrow with dysplasia of three lineages was observed. In the bone marrow cytogenic analysis, a -6, clonal cytogenic abnormality was observed. 45XY, der(6), t(6;6)(q16;q23). He was diagnosed as having chronic myelomonocytic leukemia. This is a difficult case in which it was diagnosed as refractory thrombocytopenia as a subgroup of myelodysplastic syndrome, rather than immune thrombocytopenic purpura. which might have preceded the development of chronic myelomonocytic leukemia.
Asunto(s)
Leucemia Mielomonocítica Crónica/etiología , Púrpura Trombocitopénica/complicaciones , Anciano , Anciano de 80 o más Años , Humanos , Leucemia Mielomonocítica Crónica/fisiopatología , Masculino , SíndromeRESUMEN
We previously reported the protection of hematopoietic cells from methotrexate (MTX) toxicity using an N2-based double copy vector containing serine 31 (S31)-mutated dihydrofolate reductase (DHFR) (DC/SV6S31). To examine whether the use of SFG-based dicistronic vectors will lead to improvement in gene transfer over the DC/SV6 vector, we compared the protection provided by MTX to NIH3T3 cells and hematopoietic progenitor cells infected with these retroviral constructs containing the S31 variant DHFR cDNA. In NIH3T3 cells, the 50% effective dose values of MTX conferred by the SFG vector were 8-fold higher than those obtained with the DC/SV6 vector. DHFR mRNA levels were 22-fold and 38-fold higher than that seen for the DC/SV6 vector according to Northern blot and real-time polymerase chain reaction analysis, respectively. However, DHFR protein expression and DHFR enzyme activity were only 1.5-fold and 2-fold higher in the SFG vector, respectively, indicating that the mRNA from the SFG vector is translated less efficiently than the mRNA generated from the DC/SV6 vector. Furthermore, the degree of MTX protection conferred by each vector in both mouse and human hematopoietic cells was the same. These results indicate that the in vitro transduction efficiency and transgene expression of human DHFR in hematopoietic progenitor cells is equally conferred by both vectors.
Asunto(s)
Células 3T3/efectos de los fármacos , Médula Ósea/metabolismo , ADN Complementario/genética , Resistencia a Medicamentos/genética , Células Madre Hematopoyéticas/metabolismo , Metotrexato/farmacología , Virus de la Leucemia Murina de Moloney/genética , Tetrahidrofolato Deshidrogenasa/genética , Animales , Northern Blotting , Southern Blotting , Western Blotting , Médula Ósea/efectos de los fármacos , Cartilla de ADN/química , Regulación Enzimológica de la Expresión Génica , Vectores Genéticos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/análisis , Células Madre Hematopoyéticas/efectos de los fármacos , Humanos , Leucemia/patología , Masculino , Ratones , Regiones Promotoras Genéticas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tetrahidrofolato Deshidrogenasa/metabolismo , Transducción Genética , Células Tumorales CultivadasRESUMEN
BACKGROUND: Evidence for vitamin B12 deficiency usually involves combinations of low serum vitamin B12 levels, clinical and metabolic abnormalities, and therapeutic response. Identification of the underlying cause is important in the diagnosis of vitamin B12 deficiency that is usually attributed to malabsorption. Helicobacter pylori is one of the most common causes of peptic ulcer disease worldwide and a major cause of chronic superficial gastritis leading to atrophy of gastric glands. It is suggested that there may be a casual relationship between H. pylori and food-cobalamin malabsorption. OBJECTIVES: To evaluate the H. pylori incidence in patients with vitamin B12 deficiency prospectively and to assess whether treatment for H pylori infection could correct this deficiency over time. PATIENTS AND METHODS: We performed a prospective cohort study involving 138 patients who had anemia and vitamin B12 deficiency. An upper gastrointestinal endoscopy was performed to assess the severity of atrophic gastritis and biopsy specimens for Campylobacter-like organisms tests and histological examination for H pylori were obtained at the time of diagnosis. The diagnosis of H. pylori prompted a combination treatment. RESULTS: Helicobacter pylori was detected in 77 (56%) of 138 patients with vitamin B12 deficiency and eradication of H pylori infection successfully improved anemia and serum vitamin B12 levels in 31 (40 %) of 77 infected patients. CONCLUSIONS: Helicobacter pylori seems to be a causative agent in the development of adult vitamin B12 deficiency. Eradication of H. pylori infection alone may correct vitamin B12 levels and improve anemia in this subgroup of patients.
Asunto(s)
Anemia Perniciosa/etiología , Infecciones por Helicobacter/complicaciones , Helicobacter pylori , Deficiencia de Vitamina B 12/microbiología , Adulto , Anciano , Causalidad , Endoscopía Gastrointestinal , Femenino , Mucosa Gástrica/microbiología , Mucosa Gástrica/patología , Infecciones por Helicobacter/tratamiento farmacológico , Helicobacter pylori/aislamiento & purificación , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Recurrencia , Deficiencia de Vitamina B 12/complicacionesRESUMEN
We investigated whether retrovirus-mediated transfer of the herpes simplex thymidine kinase gene into a human endometrial carcinoma (EC4) cell line can sensitize these cells to the prodrug ganciclovir (GCV) and thereby provide a therapeutic option for this cancer. A retrovirus encoding for the herpes simplex virus tip-1 (HSV) thymidine kinase (tk) gene was generated in which expression of tk is under control of the myeloproliferative sarcoma virus (MPSV) promoter/enhancer. We used human mutated dihydrofolate reductase (DHFR) cDNA as a selectable marker. Expression of tk was confirmed by Northern blot analysis and reverse transcription polymerase chain reaction. We demonstrated that the combination of retrovirally mediated tk gene transfer and GCV treatment effectively inhibits proliferation and causes death of EC4 cells in vitro. A bystander killing effect was observed when 90% of uninfected tumor cells were mixed with only 10% of HSVtk-infected cells. We suggest that a gene therapy approach to endometrial carcinoma can be established using retroviral transfer of HSVtk to tumor cells and subsequent administration of GCV.