RESUMEN
The vitamin D(3) receptor, which is the nuclear receptor for 1alpha,25-dihydroxyvitamin D(3) (VD(3)), forms a heterodimer with the retinoid X receptor (RXR), which is the nuclear receptor for 9-cis-retinoic acid (9-cis-RA). The heterodimer binds to a specific response element consisting of two directly repeated pairs of motifs, AGGTGA, spaced by three nucleotides [direct repeat (DR) 3] and modulates the expression of VD(3)-responsive genes. Telomerase activity, which is seen in most immortal cells and germ cells, is a complex of enzymes that maintain the length of telomeres. One of the major components of human telomerase, human telomerase reverse transcriptase (hTERT), is the catalytic subunit, and the expression of hTERT might correlate most strongly with telomerase activity. We found that the sequence of 5'-AGTTCATGGAGTTCA-3' (DR3') is similar to that of DR3 in the promoter region of hTERT. Our results showed that the combination of VD(3) and 9-cis-RA inhibited telomerase activity through direct interaction of the heterodimer of vitamin D(3) receptor and RXR with the DR3' sequence in the hTERT promoter as well as the combination of VD(3) and selective RXR ligand did. Also, in vivo data showed that the growth of xenografts in nude mice was inhibited by VD(3) and 9-cis-RA. The results of the present study provide evidence on the molecular mechanism of the inhibition of cell growth by these agents, and they could be novel therapeutic agents for prostate cancer.
Asunto(s)
Calcitriol/farmacología , Neoplasias de la Próstata/genética , Telomerasa/metabolismo , Tretinoina/farmacología , Alitretinoína , Animales , Antineoplásicos/farmacología , Proteínas de Unión al ADN , Sinergismo Farmacológico , Humanos , Masculino , Ratones , Regiones Promotoras Genéticas/efectos de los fármacos , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/enzimología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Telomerasa/antagonistas & inhibidores , Telomerasa/genética , Transcripción Genética/efectos de los fármacos , Células Tumorales CultivadasRESUMEN
PURPOSE: We report our experience in 10 years of sperm cryopreservation to reveal the present state of the cryopreservation project. MATERIALS AND METHODS: 42 germ cell tumor, 110 non-germ cell tumor and 2 non-malignant disease patients who visited our clinic for semen cryopreservation were retrospectively analyzed. RESULTS: Only 7 (20%) out of 35 unilateral testicular tumor patients who had received no chemotherapy met the WHO criteria for sperm concentration and motility. However, there were no patients with azoospermia. Three testicular tumor patients with previous chemotherapy and 4 retroperitoneal germ cell tumor patients had poor sperm concentrations and motilities. Twenty (52%) out of 38 non-germ cell tumor patients without previous chemotherapy met the WHO criteria. In contrast, only 9 patients (13%) met the WHO criteria among 72 patients with previous chemotherapy. Twenty-nine patients (40%) with chemotherapy were azoospermia. Totally, 74 (96%) of 77 tumor patients' semen without chemotherapy and 34 (45%) of 75 with chemotherapy were cryopreserved. Sperm from a patient with testicular torsion were cryopreserved. CONCLUSION: Most cancer patients without previous chemotherapy, regardless of underlying disease, had abundant motile sperm. However, half of the patients who had received chemotherapy did not have suitable sperm for freezing. It is important to inform young cancer patients of the cryopreservation project immediately after the diagnosis is made.
Asunto(s)
Criopreservación , Enfermedades de los Genitales Masculinos , Neoplasias de los Genitales Masculinos , Preservación de Semen , Adolescente , Adulto , Criopreservación/estadística & datos numéricos , Humanos , Masculino , Persona de Mediana Edad , Preservación de Semen/estadística & datos numéricos , Recuento de Espermatozoides , Motilidad EspermáticaRESUMEN
Recent studies have demonstrated that GnRH-analogues can stimulate regeneration of spermatogenesis of rats when administered after testicular damages. Although the mechanism of this phenomenon has not been elucidated yet, stem cell factor (SCF) produced by Sertoli cells was proposed to mediate the effects of GnRH-analogues on spermatogonial proliferation and/or survival. In the present study, we quantitatively evaluated the proliferation of spermatogonia and addressed whether SCF mediates the effect of GnRH-analogue on spermatogonial proliferation, using a novel approach combining spermatogonial transplantation and laser confocal microscopic observation. In the first experiment, using wild-type mice as recipients for spermatogonial transplantation, the number of donor spermatogonia per 100 Sertoli cells in each spermatogenic colony was significantly higher in the experimental group of mice treated with leuprorelin, a GnRH-agonist, than that of the control group at 4 and 5 wk after transplantation. In the second experiment, Steel/Steeldickie (Sl/Sld) mutant mice, which lack expression of membrane bound form SCF, were used as recipients. As seen in the first experiment, the number of undifferentiated spermatogonia was significantly higher in leuprorelin-treated than in the control group. Since undifferentiated spermatogonia do not express the receptor of SCF, the present study clearly demonstrates that neither membrane-bound nor secreted forms of SCF are involved in the mechanism of GnRH-analogue's effect on spermatogonial proliferation and/or survival.
Asunto(s)
Hormona Liberadora de Gonadotropina/análogos & derivados , Hormona Liberadora de Gonadotropina/farmacología , Proteínas Proto-Oncogénicas c-kit/fisiología , Transducción de Señal/fisiología , Espermatogonias/efectos de los fármacos , Factor de Células Madre/genética , Factor de Células Madre/fisiología , Animales , Peso Corporal/efectos de los fármacos , Células Germinativas/efectos de los fármacos , Proteínas Fluorescentes Verdes , Inmunohistoquímica , Leuprolida/farmacología , Proteínas Luminiscentes/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microscopía Confocal , Tamaño de los Órganos/efectos de los fármacos , Túbulos Seminíferos/fisiología , Recuento de Espermatozoides , Espermatogénesis/genética , Espermatogonias/trasplanteRESUMEN
It has been documented that renal cell carcinomas (RCCs) occur frequently in patients treated with long-term dialysis, especially in cases of end-stage renal disease (ESRD)/acquired cystic disease of the kidney (ACDK). To address the molecular pathogenesis of ESRD/ACDK-associated RCCs, we examined 14 RCCs (7 clear-cell and 7 papillary carcinomas) in patients receiving dialysis for somatic mutations of the von Hippel-Lindau disease (VHL) gene as well as of the tyrosine kinase domain of the MET oncogene. Direct sequencing analyses revealed that three tumors exhibited VHL frameshifts (618delA, 386-395del10-bp, and 723-724insTC). One of the VHL mutated tumors showed additional loss of heterozygosity at the VHL gene locus. Histopathologic and clinical data demonstrated that the three tumors having VHL mutations were clear-cell RCCs occurring in ESRD with 55, 106, and 156 months of dialysis history, respectively. We did not find any tumors with mutations in the tyrosine kinase domain of the MET. These results demonstrated that the VHL tumor-suppressor gene is also involved in a subset of clear-cell RCCs occurring in ESRD/ACDK, as in the case of sporadic clear-cell RCCs. However, mutations of the MET oncogene could not be found in the seven ESRD/ACDK-associated papillary RCCs examined.
Asunto(s)
Adenocarcinoma de Células Claras/genética , Carcinoma de Células Renales/genética , Enfermedades Renales Quísticas/genética , Fallo Renal Crónico/genética , Neoplasias Renales/genética , Ligasas/genética , Mutación/genética , Proteínas Supresoras de Tumor , Ubiquitina-Proteína Ligasas , Enfermedad de von Hippel-Lindau/genética , Adulto , Anciano , Carcinoma Papilar/genética , Femenino , Genes Supresores de Tumor , Humanos , Enfermedades Renales Quísticas/terapia , Fallo Renal Crónico/terapia , Pérdida de Heterocigocidad/genética , Masculino , Persona de Mediana Edad , Factores de Tiempo , Proteína Supresora de Tumores del Síndrome de Von Hippel-LindauRESUMEN
PURPOSE: Cryptorchidism is an adverse condition of spermatogenesis in many mammals. Surgical cryptorchidism in rats lasting more than a few weeks is so detrimental that spermatogenesis cannot be completely recovered even after orchiopexy. We evaluated the efficacy of the high dose gonadotropin-releasing hormone (Gn-RH) agonist leuprorelin acetate on damaged spermatogenesis in rat cryptorchid testes. MATERIALS AND METHODS: Male Fisher rats were divided into 2 groups of 6 each and bilateral cryptorchidism was artificially produced. Five weeks later all rats underwent bilateral orchiopexy. One group served as the control, while the other received Gn-RH agonist injections at orchiopexy and 4 weeks later. The animals were sacrificed 15 weeks after orchiopexy. The weight of the body, testis and epididymis was measured and the histology of spermatogenesis was examined. For statistical analysis the Student t test was applied. RESULTS: Testes in the Gn-RH group rats showed significant recovery of spermatogenesis up to complete spermatozoa formation, while those in control rats remained almost degenerated. The mean incidence of seminiferous tubules with recovered spermatogenesis plus or minus standard deviation was significantly higher in the Gn-RH than in the control group (87.8% +/- 6.0% versus 12.5% +/- 7.7%, p <0.001). CONCLUSIONS: Administering the high dose Gn-RH agonist leuprorelin acetate after orchiopexy greatly enhanced the recovery of spermatogenesis in rats. This finding is in accordance with other recent reports that treatment with Gn-RH analogues promotes the regeneration of once damaged spermatogenesis. On the other hand, these findings may cause one to question supplementation therapy now used in regular practice to boys with cryptorchidism.
Asunto(s)
Criptorquidismo/cirugía , Leuprolida/administración & dosificación , Espermatogénesis/efectos de los fármacos , Testículo/cirugía , Animales , Criptorquidismo/fisiopatología , Epidídimo/patología , Células Germinativas/patología , Masculino , Tamaño de los Órganos , Ratas , Ratas Endogámicas F344 , Túbulos Seminíferos/patología , Testículo/patologíaRESUMEN
One hundred and twenty-nine patients with benign prostatic hypertrophy (BPH) were registered and treated with allylestrenol. Allylestrenol was administered at a dose of 50 mg/day given twice a day for 16 weeks. Out of 129 patients with a mean age of 67.8 years old, 92 cases completed the study and 48 cases with moderate symptoms were objectively evaluated with "Criteria for Treatment Efficacy in BPH" proposed by The Japanese Urological Association in 1997. Prostate volume was significantly decreased from 32.7 +/- 11.9 to 27.4 +/- 11.2 ml (mean +/- SD), and maximum flow rate was significantly increased from 8.4 +/- 3.4 to 10.8 +/- 5.0 ml/sec. Residual urine volume was significantly decreased from 62.4 +/- 57.4 to 37.0 +/- 38.7 ml. IPSS was significantly decreased from 15.3 +/- 4.9 to 9.9 +/- 4.0, and QOL index was significantly decreased from 4.4 +/- 0.8 to 2.7 +/- 1.2. The efficacy of allylestrenol was shown by its effects on prostate volume (anatomy), maximum urinary flow rate (function), and symptom scores (symptom) at the end of 16 weeks of treatment. The rates of improvement for symptoms, QOL, function, and anatomy are 68.7% (N = 48), 79.2% (N = 48), 50.0% (N = 48), and 61.0% (N = 41), respectively. Overall efficacy (Good and Fair) was 70.9% (N = 48). During this study, 5 patients (3.9%) complained of loss of libido and 2 patients dropped out. In conclusion, allylestrenol was demonstrated to be a quite effective and safe medical treatment for patients with symptomatic BPH based on the criteria for treatment efficacy in BPH.
Asunto(s)
Alilestrenol/uso terapéutico , Congéneres de la Progesterona/uso terapéutico , Hiperplasia Prostática/tratamiento farmacológico , Anciano , Anciano de 80 o más Años , Esquema de Medicación , Humanos , Masculino , Persona de Mediana Edad , Guías de Práctica Clínica como Asunto , Hiperplasia Prostática/fisiopatología , Calidad de Vida , Sistema de RegistrosRESUMEN
PURPOSE: To assess the effect of high concentration of sodium and potassium ion on the motilities of human sperm following the preservation in electrolyte-free solution. PATIENT: Semen samples were obtained from patients attending our infertility clinic. The motilities of their sperm were more than 40%. METHOD: Ejaculated sperm were washed by using the electrolyte-free Percoll gradient and then preserved in electrolyte-free solution at 4 degrees C for a week. The preserved sperm was incubated and analyzed after the addition of electrolyte solutions under acid or alkaline condition. The sperm were continuously preserved for a day at room temperature and analyzed. RESULT: The sperm was by the addition of 35-135 mM Na+ or 5-135 mM K+ in acid solutions. Under the alkaline conditions, sperm were reactivated without Na+ or K+. However, the addition of 135 mM Na+ or K+ significantly decreased in sperm motilities. The some sperm in the solution containing low concentration of Na+ or K+ ions maintained their motilities for a day at room temperature. However, all sperm lost their motilities in the 135 mM Na+ or K+ solutions. CONCLUSION: The high concentration of Na+ or K+ ions had detrimental effects on motilities of sperm following the preservation in electrolyte-free solution.
Asunto(s)
Potasio/farmacología , Preservación de Semen , Sodio/farmacología , Motilidad Espermática/efectos de los fármacos , Humanos , Masculino , Soluciones Preservantes de Órganos , Preservación de Semen/métodosRESUMEN
To delineate more precisely the somatic von Hippel-Lindau disease (VHL) gene alteration as well as to elucidate its etiologic role in renal tumorigenesis, we examined a total of 240 sporadic renal cell carcinomas (RCCs) for somatic VHL gene alterations by DNA-SSCP followed by sequencing, methylation-specific PCR assay, microsatellite LOH study, and Southern blot analysis. Intragenic mutation of the VHL gene was found exclusively in clear-cell or variant-type RCCs at a frequency of 51% (104/202). Hypermethylation of the VHL promoter region was detected in an additional 11 clear-cell RCCs. Microsatellite analysis demonstrated that LOH of the VHL locus was found in 140/155 (90%) informative clear-cell RCCs. The VHL gene therefore seems to be inactivated in a two-hit manner by intragenic mutation or hypermethylation plus allelic loss in clear-cell RCC. Genomic rearrangement of the VHL gene detected by Southern analysis was not found (0/216 cases); this is in contrast to germ lines in which Southern aberrations consisted of 7-19% of the mutations. Clinicopathologic data demonstrated that VHL mutation/LOH did not vary according to tumor progression in clear-cell RCC, including tumor diameter, stage, grading, distant metastasis, and lymph node metastasis. Interestingly, VHL mutation was significantly less frequent in RCCs occurring in younger (< or = 55 years) than that in older (> or = 56 years) patients. These data suggested that the inactivation of the VHL tumor-suppressor gene is a specific genetic change in clear-cell RCC, and that it may occur at an early or first step in the clear-cell tumorigenic pathway rather than as a late event.