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INTRODUCTION: Vascular management of the right renal vein during laparoscopic living donor nephrectomy is still an unsolved problem. This short vessel has limited the use of right kidneys. However, the right kidney should be harvested in some instances. Based on experience in open donor nephrectomy, our unit has used the donor gonadal vein to obtain a longer renal vein in this setting. METHODS: Four consecutive living related donors with the indication for laparoscopic right nephrectomy underwent this procedure. Three donors were females and the overall average age was 48.5 years. The renal vein was controlled with a 30-mm stapler and we included 5-6 cm of the ipsilateral gonadal vein during the harvest. The donor kidney was perfused and renal vessels prepared under cold conditions. The gonadal vein was opened longitudinally and sutured to the donor right renal vein as a wide tube in 3 cases and as a spiral tube in 1 case with 6-0 monofilament suture. RESULTS: This procedure extended the bench work between 25 to 40 minutes permitting an 2.5- to 3.5-cm extension of the donor vein. The transplantations were performed in the usual mode and the vein enlargement enormously facilitated the implantation surgery. All recipients displayed immediate graft function; no complications were observed with this strategy. CONCLUSIONS: Vein extension with the gonadal vein was a simple, safe method to enlarge the renal vein among right living donor kidneys procured using laparoscopy.
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Laparoscopía/métodos , Donadores Vivos , Nefrectomía/métodos , Venas Renales/cirugía , Recolección de Tejidos y Órganos/métodos , Familia , Femenino , Lateralidad Funcional , Humanos , Masculino , Persona de Mediana Edad , Arteria Renal/anomalías , Arteria Renal/cirugía , SuturasRESUMEN
OBJECTIVE: Monitoring of cyclosporine (CsA) is critical during the induction of immunosuppressive therapy. Although most centers have incorporated C2 levels, our unit still uses an abbreviated AUC model which includes concentrations at C1, C2, and C6 post-dose (AUC(1-6)). The objective of this study was to compare both strategies of CsA monitoring during the first 30 days after kidney transplantation. PATIENTS AND METHODS: The study included 89 recipients induced with CsA microemulsion and steroids. AUC(1-6) profiles were performed around days 3, 10, and 30 after transplantation with a target of 5500 to 6000 ng*h/mL considered therapeutic. For comparison purposes, a value of C2 >/= 1500 ng/mL was also considered therapeutic. Mean C2 and AUC(1-6) values were low dated with biopsy-proven acute rejection episodes (BPAR) during the study period. RESULTS: Twenty patients received living donor kidneys and overall there were 46 females. During this period, 253 AUC(1-6) were performed including 44 (17.4%) below the therapeutic range. When the analysis included only C2, 171 (67.6%) were below the therapeutic target (P < .001). Five patients experience BPAR and only AUC(1-6) at day 10 discriminated rejectors versus nonrejectors (5645 +/- 1390 and 8221 +/- 2502, respectively; P = .008). C2 was not significantly different at any time in either group. CONCLUSIONS: In this study, abbreviated AUC monitoring more adequately identified patients at risk for acute rejection than C2. Recommended C2 concentration levels need to be redefined in our patients.
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Ciclosporina/farmacocinética , Ciclosporina/uso terapéutico , Rechazo de Injerto/inmunología , Trasplante de Riñón/inmunología , Corticoesteroides/uso terapéutico , Adulto , Área Bajo la Curva , Cadáver , Relación Dosis-Respuesta a Droga , Emulsiones , Femenino , Rechazo de Injerto/epidemiología , Rechazo de Injerto/prevención & control , Supervivencia de Injerto/inmunología , Prueba de Histocompatibilidad , Humanos , Inmunosupresores/farmacocinética , Inmunosupresores/uso terapéutico , Donadores Vivos , Masculino , Persona de Mediana Edad , Monitoreo Fisiológico/métodos , Selección de Paciente , Estudios Retrospectivos , Donantes de Tejidos , Adulto JovenRESUMEN
OBJECTIVE: The purpose of this study was to evaluate the importance of various factors on 1-year serum creatinine (SCr) as a surrogate endpoint for allograft survival among a series of kidney transplantations performed at 2 centers. PATIENTS AND METHODS: Two hundred sixty consecutive renal transplantations were included with overall mean age of 40 +/- 13 years, including 55% men and 19% living donor grafts. Factors considered for analysis were donor and recipient ages, and sexes, number of transplantations, panel-reactive antibodies, total number of HLA mismatches, cold ischemia time (CIT), acute rejection (AR) rate, and presence/duration of delayed graft function (DGF). Multiple regression analyses were performed for 1-year SCr, AR rate, and DGF duration. RESULTS: One-year SCr was 1.46 +/- 0.5 mg/dL, 6-month AR rate was 22%, and DGF rate was 29% of mean duration 3 +/- 8 days. Multiple regression analysis for lower 1-year SCr value identified as significant female recipient sex, lower donor age, absence of AR within 6 months, and decreased DGF duration (P < .05). The only significant factor affecting AR rate was DGF duration. Finally, prolonged CIT was associated with a longer DGF duration. CONCLUSIONS: We confirmed that 1-year SCr was primarily affected by well-known factors, such as AR incidence, donor age, and female recipient sex. However, we identified DGF duration as a significant factor affecting 1-year SCr. AR rate was also associated with DGF duration, which in turn depended upon longer CIT.
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Creatinina/sangre , Supervivencia de Injerto/fisiología , Trasplante de Riñón/fisiología , Adulto , Cadáver , Femenino , Estudios de Seguimiento , Rechazo de Injerto/epidemiología , Rechazo de Injerto/inmunología , Antígenos HLA/inmunología , Prueba de Histocompatibilidad , Humanos , Isoanticuerpos/inmunología , Trasplante de Riñón/inmunología , Donadores Vivos , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Factores de Tiempo , Donantes de TejidosRESUMEN
BACKGROUND: Nuclear accumulation of p53 protein has been shown to be strongly associated with missense p53 mutations. Studies of nuclear accumulation of p53 protein in prostate carcinoma cells have to date been confined to material from primary tumors. PURPOSE: We studied the accumulation of p53 protein in specimens obtained from primary and metastatic sites of prostate carcinoma. By examining the accumulation of this protein as a function of stage, histologic grade, and androgen responsiveness of the tumor, we hoped to determine the role of p53 mutation in the progression of prostate carcinoma. METHODS: The accumulation of the p53 protein in the cell nuclei was determined by immunohistochemical methods using polyclonal antibody to human p53 CM-1. The material studied consisted of formalin-fixed, paraffin-embedded tissue obtained from primary tumors and metastases of 92 patients with prostate carcinoma. Twelve samples from 11 patients were analyzed for the presence of mutations within exons 5-8 of the p53 gene (also known as TP53) by polymerase chain reaction-single-stranded conformation polymorphism (PCR-SSCP) analysis. Sequence analysis was subsequently performed on DNA obtained by polymerase chain reaction amplification of PCR-SSCP reactions produced from six different specimens. The chi-square test, Fisher's exact test, and the Freeman Halton test were used for statistical analyses of the results. RESULTS: All tumors with p53 accumulation were metastatic (stage D), poorly differentiated, and androgen independent. Nuclear accumulation of p53 protein was strongly associated with stage (D2 versus D1 versus A-C, P < .0001), grade (Gleason score 8-10 versus 5-7, P < .003), and androgen sensitivity (androgen independent versus dependent, P < .0001). Logistic regression analysis demonstrated that androgen sensitivity predicted p53 outcome better than did stage (P < .0001) or grade alone (P < .006). There was a perfect concordance between the results obtained by PCR-SSCP analysis and the p53 protein accumulation determined by immunohistochemistry in the 12 samples studied. Mutation of the p53 gene was confirmed by sequencing DNA obtained from six specimens positive in the PCR-SSCP assay. CONCLUSIONS: p53 gene mutation is a late event in the progression of prostate cancer and is associated with advanced (metastatic) stage, loss of differentiation, and the transition from androgen-dependent to androgen-independent growth. IMPLICATION: Testing of prostate cancer biopsy specimens from metastatic sites for p53 protein accumulation and gene mutation may provide useful prognostic information and could influence the recommended course of treatment.
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Carcinoma/genética , Carcinoma/metabolismo , Genes p53/genética , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Secuencia de Bases , Carcinoma/patología , Carcinoma/secundario , Distribución de Chi-Cuadrado , Humanos , Modelos Logísticos , Metástasis Linfática , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Mutación , Neoplasias Hormono-Dependientes/genética , Neoplasias Hormono-Dependientes/metabolismo , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Hiperplasia Prostática/metabolismo , Neoplasias de la Próstata/patologíaRESUMEN
We studied loss of heterozygosity (LOH) on human chromosome 7q to determine the location of a putative tumor suppressor gene (TSG) in human primary prostate carcinomas. Samples were obtained from 16 primary prostate carcinomas surgically removed from patients at The University of Texas M. D. Anderson Cancer Center. Paired normal and tumor DNAs were used as template for PCR amplification of a set of 14 CA microsatellite repeats on 7q21-qter. Twelve of 16 cases studied had LOH at one or more loci on 7q. Eighty-three percent LOH (five of six informative cases) was detected with D7S522 at 7q31.1-7q31.2. Percentage of LOH was normally distributed around D7S522. The high incidence of LOH in primary prostate carcinomas suggests that there is a TSG relevant to the development of prostate cancers at 7q31.1-31.2, confirming our previous functional evidence for a TSG at this location. Further research needs to be conducted to establish the identity and function of this putative TSG.
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Carcinoma/genética , Deleción Cromosómica , Cromosomas Humanos Par 7/genética , Genes Supresores de Tumor , Neoplasias de la Próstata/genética , Mapeo Cromosómico , ADN Satélite/genética , Humanos , MasculinoRESUMEN
Although gallbladder carcinoma (GBC) is a highly malignant neoplasm, there is very limited information about the molecular changes involved in its pathogenesis. To identify the chromosomal locations of putative tumor suppressor gene loci involved in the pathogenesis of GBC, we conducted a genome-wide allelotyping or loss of heterozygosity (LOH) analysis of GBCS: Microdissected tissue from 24 archival GBCs and their matched control DNAs were analyzed for PCR-based LOH using 169 microsatellite markers spanning all nonacrocentric autosomal arms and the X chromosome. The chromosomal arms with the greatest frequencies of LOH (> or = 60%) were 3p, 6q, 7q, 8p, 9p, 9q, 11q, 12q, 17p, 18q, 19p, 22q, and XQ: The average fractional allele loss index in GBC cases was high (0.43) and frequent breakpoints were detected in gallbladder tumors. Of interest, 21 different regions of frequent LOH (hot spots) defined as > or = 50% for individual GBC samples were detected in this neoplasm, nearly half of them confined to one microsatellite marker. We conclude that in GBC at least 21 chromosomal regions with frequent allele losses are involved, suggesting that several putative tumor suppressor genes are inactivated in its pathogenesis. Overall, these data provide global estimates of the extent of genetic changes leading to GBC and will be useful for the identification of new tumor suppressor genes and for multiple new markers for translational research.
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Adenocarcinoma/genética , Neoplasias de la Vesícula Biliar/genética , Pérdida de Heterocigocidad , Anciano , Anciano de 80 o más Años , Rotura Cromosómica , Femenino , Eliminación de Gen , Genoma Humano , Humanos , Masculino , Repeticiones de Microsatélite/genética , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Cromosoma XRESUMEN
The significance of apoptosis in relation to the development and progression of prostate cancer remains largely undefined. bcl-2 is an oncogene that functions by overriding apoptosis. bcl-2 expression was localized to the basal epithelial cells in the normal human prostate with the use of immunohistochemistry. Androgen-dependent and androgen-independent prostate carcinomas were evaluated immunohistochemically for bcl-2 expression. bcl-2 was undetectable in 13 of 19 cases of androgen-dependent cancers. In contrast, androgen-independent cancers displayed diffuse, high levels of bcl-2 staining (P < 0.01). In rats, steady-state levels of bcl-2 mRNA, assessed by S1 assays, reached maximum levels 10 days following castration. Addition of exogenous testosterone with, or without, flutamide demonstrated that the increased bcl-2 mRNA resulted from androgen ablation. Our findings indicate that bcl-2 expression is augmented following androgen ablation and is correlated with the progression of prostate cancer from androgen dependence to androgen independence.
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Expresión Génica , Próstata/metabolismo , Neoplasias de la Próstata/etiología , Proteínas Proto-Oncogénicas/genética , Proto-Oncogenes , Andrógenos/farmacología , Animales , Apoptosis , Masculino , Neoplasias Hormono-Dependientes/etiología , Neoplasias de la Próstata/genética , Proteínas Proto-Oncogénicas c-bcl-2 , Ratas , Ratas Sprague-DawleyRESUMEN
Cell adhesion molecules have been suggested to function as tumor suppressor molecules. We have been studying one of the epithelial cell adhesion molecules (C-CAM), which belongs to the immunoglobulin gene superfamily. Transfection of a C-CAM cDNA expression vector into a highly tumorigenic human prostate cancer cell line (PC-3) suppresses tumor formation in nude mice. Alternatively, reducing C-CAM expression levels in the nontumorigenic rat prostate epithelial cell line NbE by the antisense expression vector markedly increases tumorigenicity of NbE cells in nude mice. These results suggest that C-CAM may be a tumor suppressor in prostate cancer. In this study, we examined the relationship between C-CAM expression during human prostate development and neoplastic progression by immunohistochemical staining of frozen sections. C-CAM predominantly localized on the plasma membrane of the basal cell layer in both the fetal and normal adult prostate gland. However, an overall decreased staining was seen in benign prostatic hyperplasia and high grade prostatic intraepithelial neoplasia. Furthermore, C-CAM was not detected in prostate carcinomas. Thus, a decrease in C-CAM expression may be an early event in hyperplastic/neoplastic transformation. These observations support the suggestion that C-CAM is a tumor suppressor in prostate cancer progression.
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Antineoplásicos/análisis , Moléculas de Adhesión Celular/análisis , Próstata/química , Próstata/embriología , Neoplasias de la Próstata/química , Moléculas de Adhesión Celular/fisiología , Epitelio/química , Homeostasis , Humanos , Masculino , Hiperplasia Prostática/metabolismoRESUMEN
A candidate tumor suppressor gene, MMAC1/PTEN, located in human chromosome band 10q23, was recently identified based on sequence alterations observed in several glioma, breast, prostate, and kidney tumor specimens or cell lines. To further investigate the mutational profile of this gene in human cancers, we examined a large set of human tumor specimens and cancer cell lines of many types for 10q23 allelic losses and MMAC1 sequence alterations. Loss of heterozygosity (LOH) at the MMAC1 locus was observed in approximately one-half of the samples examined, consistent with the high frequency of 10q allelic loss reported for many cancers. Of 124 tumor specimens exhibiting LOH that have been screened for MMAC1 alterations to date, we have detected variants in 13 (approximately 10%) of these primary tumors; the highest frequency of variants was found in glioblastoma specimens (approximately 23%). Novel alterations identified in this gene include a missense variant in a melanoma sample and a splicing variant and a nonsense mutation in pediatric glioblastomas. Of 76 tumor cell lines prescreened for probable LOH, microsequence alterations of MMAC1 were detected in 12 (approximately 16%) of the lines, including those derived from astrocytoma, leukemia, and melanoma tumors, as well as bladder, breast, lung, prostate, submaxillary gland, and testis carcinomas. In addition, in this set of tumor cell lines, we detected 11 (approximately 14%) homozygous deletions that eliminated coding portions of MMAC1, a class of abnormality not detected by our methods in primary tumors. These data support the occurrence of inactivating MMAC1 alterations in multiple human cancer types. In addition, we report the discovery of a putative pseudogene of MMAC1 localized on chromosome 9.
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Cromosomas Humanos Par 10 , Mutación , Neoplasias/genética , Monoéster Fosfórico Hidrolasas , Proteínas Tirosina Fosfatasas/genética , Proteínas Supresoras de Tumor , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patología , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Niño , Mapeo Cromosómico , Exones , Femenino , Eliminación de Gen , Marcadores Genéticos , Variación Genética , Glioblastoma/genética , Glioblastoma/patología , Glioma/genética , Glioma/patología , Humanos , Intrones , Neoplasias Renales/genética , Neoplasias Renales/patología , Masculino , Neoplasias/patología , Fosfohidrolasa PTEN , Mutación Puntual , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/patología , Proteínas Tirosina Fosfatasas/análisis , Proteínas Tirosina Fosfatasas/biosíntesis , Eliminación de Secuencia , Neoplasias Testiculares/genética , Neoplasias Testiculares/patología , Células Tumorales CultivadasRESUMEN
We studied loss of heterozygosity (LOH) on human chromosome 13q in prostate cancer specimens to determine the location of a putative tumor suppressor gene (TSG) and to correlate these losses with the clinicopathological stage of the disease. Overall 13 (21%) of 61 specimens analysed had an allele loss on the long arm of chromosome 13. The most frequent (37%) LOH among the informative cases with allele losses was detected at the D13S284 locus on chromosome 13q14. 3. A portion of the DNA segment that spans this locus and is flanked by the microsatellite loci D13S153 and D13S163 was lost in 85% of the specimens with allele losses and was designated as a LOH cluster region (LCR). The LCR spans more than 6 Mbp of DNA. The results suggest that a TSG relevant for the development of prostate cancer is located telomeric to the RB locus. There was a significant correlation (P=0.0024) between chromosome 13q LOH and advanced metastatic disease, suggesting that loss of 13q14.3 region is associated with prostate cancer progression. However, further research must be conducted to establish the identity and function of this putative TSG.
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Cromosomas Humanos Par 13 , Genes Supresores de Tumor , Pérdida de Heterocigocidad , Neoplasias de la Próstata/genética , Mapeo Cromosómico , Humanos , Masculino , Repeticiones de MicrosatéliteRESUMEN
We studied loss of heterozygosity (LOH) on the long arm of human chromosome 18 in prostate cancer to determine the location of a putative tumor suppressor gene (TSG) and to correlate these losses with the pathological grade and stage of the cancer. Of 48 specimens analysed 17 (35.4%) lost at least one allele on chromosome 18q. All the specimens with allelic losses lost at least one allele within chromosomal region 18q21. Allelic losses picked at D18S51 (19%) and D18S858 (17%). A 0.58 cM DNA segment that includes the D18S858 locus and is flanked by the microsatellite loci D18S41 and D18S381, was lost in eight (47%) of 17 specimens with allelic losses. This segment was designated as a LOH cluster region 1 (LCR 1). Although Smad2 resides within LCR 1, it was not mutated in any of the six prostate cell lines (five prostate cancer cell lines and one immortalized prostate epithelial cell line) analysed, suggesting that it is not a candidate TSG in prostate cancer. A second LCR at 18q21, LCR 2, includes the D18S51 locus and is flanked by the D18S1109 and D18S68 loci, which are separated by 7.64 cM. LCR 2 was lost in six (35%) of the 17 specimens with chromosome 18q losses. These results suggest that chromosome 18q21 may harbor two candidate prostate cancer TSGs. The candidate TSGs DCC and Smad4 are located centromeric to the LCRs. No alleles were lost within or in close proximity to these genes, suggesting that they are not targets for inactivation by allelic losses in prostate cancer. Although there was no obvious correlation between chromosome 18q LOH and the pathological grade or stage, three (37.5%) of eight low-grade cancers and nine (32.1%) of 28 organ-confined cancers lost alleles at 18q21, suggesting that allelic losses are relatively early events in the development of invasive prostate cancer.
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Cromosomas Humanos Par 18/genética , Genes Supresores de Tumor , Pérdida de Heterocigocidad , Neoplasias de la Próstata/genética , Mapeo Cromosómico , Proteínas de Unión al ADN/genética , Silenciador del Gen , Humanos , Masculino , Repeticiones de Microsatélite/genética , Mutación , Estadificación de Neoplasias , Neoplasias de la Próstata/patología , Proteína Smad2 , Transactivadores/genéticaRESUMEN
Frequent loss of an allele at specific chromosomal regions implicates these regions as sites of tumor suppressor genes (TSG) that become inactivated during tumor progression. We have studied chromosome 8p allele losses in 32 primary human prostate carcinomas with 16 polymorphic microsatellite sequences. Overall, 22 of 32 (69%) informative specimens showed loss of allele in at least one locus. The most frequent losses of heterozygosity (LOH) occurred at the LPL locus (46%) on chromosome 8p22 and at the D8S360 (45%) and NEFL (43%) loci on chromosome 8p21. Homozygous deletions were detected at the LPL and NEFL loci at 8p22 and 8p21, respectively. The minimal region with frequent LOH and homozygous deletion, around the LPL locus, was restricted between the MSR locus and the D8S258 marker, separated by less than 9 cM. The second region was restricted between markers D8S1128 and D8S131 separated by 12 cM. The results suggest the existence of two chromosome 8p sites for candidate TSGs in prostate cancer.
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Cromosomas Humanos Par 8/genética , Eliminación de Gen , Genes Supresores de Tumor , Neoplasias de la Próstata/genética , Alelos , Sitios de Unión , Homocigoto , Humanos , Masculino , Neoplasias de la Próstata/patologíaRESUMEN
PURPOSE: We assessed the feasibility and efficacy of integrating chemotherapy and androgen ablation with radical prostatectomy in patients with locally advanced prostate cancer. The neoadjuvant approach was adopted because it allows an in situ assessment of antitumoral activity. PATIENTS AND METHODS: Thirty-three patients were enrolled who met the clinical criteria of stage T1-2, Gleason score of >/= 8 or T2b-T2c, Gleason score of 7 and prostate-specific antigen (PSA) level greater than 10 ng/mL (n = 15), or clinical stage T3 (n = 18). Therapy consisted of 12 weeks of ketoconazole and doxorubicin alternating with vinblastine, estramustine, and androgen ablation followed by prostatectomy. The ability of neoadjuvant chemotherapy and hormonal therapy to induce a 20% rate of pT0 in the prostatectomy specimen as well as surgical feasibility were assessed. RESULTS: Chemotherapy complications were comparable to those reported with this regimen previously. No major intraoperative complications occurred. Postoperative complications occurred in 10 (33%) of 30 patients. One patient died at home after discharge (postoperative day 17; no autopsy was performed). Ten (33%) of the 30 patients had organ-confined disease, and 20 (70%) of 30 had extraprostatic extension; 11 (37%) of the 30 had positive lymph nodes. Only five (17%) of 30 exhibited positive surgical margins. All patients achieved an undetectable PSA level postoperatively, and 20 of the surviving 29 patients remain without disease recurrence with a median follow-up of 13 months (range, 9 to 18 months). CONCLUSION: Chemotherapy and androgen ablation followed by radical prostatectomy was feasible in patients with locally advanced prostate cancer. Although the goal of achieving a 20% rate for pT0 status was not achieved, we believe this type of integrated therapeutic strategy should be investigated further for its ability to alter the course of regionally advanced prostate cancer.
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Antineoplásicos Hormonales/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Terapia Neoadyuvante , Prostatectomía , Neoplasias de la Próstata/terapia , Adulto , Anciano , Antagonistas de Andrógenos/administración & dosificación , Andrógenos/metabolismo , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Terapia Combinada , Doxorrubicina/administración & dosificación , Estramustina/administración & dosificación , Estudios de Factibilidad , Estudios de Seguimiento , Humanos , Cetoconazol/administración & dosificación , Masculino , Persona de Mediana Edad , Antígeno Prostático Específico/análisis , Neoplasias de la Próstata/diagnóstico por imagen , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/cirugía , Ultrasonografía , Vinblastina/administración & dosificaciónRESUMEN
PURPOSE: Prostate cancer progression is associated with deregulation of genes like E-cadherin, p21/WAF1, MMP-2, VEGF, and IGF-binding protein, 3 and 5, all of which are target genes for the transcription factor activator protein 2alpha (AP-2alpha). We, therefore, hypothesize that the development/progression of prostate cancer is associated with changes in the expression of AP-2alpha. EXPERIMENTAL DESIGN: We used immunofluorescent staining to assess the presence of AP-2alpha in normal, benign, and malignant human prostate tissues and to correlate its expression with tumor grade and stage. RESULTS: We found that although AP-2alpha was expressed in normal prostate epithelium, it was not expressed in 30 prostate cancer specimens of different Gleason scores. Moreover, AP-2alpha protein was present in the luminal cell layer but not in the basal cell layer of the normal epithelium, which indicated that the loss of AP-2alpha staining in the prostate cancer specimens was not attributable to a lack of AP-2alpha-expressing cells. Further analysis demonstrated the presence of AP-2alpha in 2 (40%) of 5 atrophic normal epithelium, in 4 (24%) of 17 cases of benign prostatic hyperplasia, and in 2 (13%) of 13 cases of high-grade prostatic intraepithelial neoplasia. Loss or reduction in AP-2alpha expression was also observed in LNCaP, LNCaP-LN3, and PC3M-LN4 cell lines. CONCLUSIONS: Our data demonstrate that AP-2alpha expression is associated with normal luminal differentiation and that a loss of AP-2alpha expression occurs early in the development of prostate adenocarcinoma. Loss of AP-2alpha may lead to deregulation in AP-2alpha target genes that normally regulate cellular growth and differentiation.
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Biomarcadores de Tumor/análisis , Diferenciación Celular/fisiología , Proteínas de Unión al ADN/análisis , Neoplasias de la Próstata/patología , Factores de Transcripción/análisis , Carcinoma/patología , Progresión de la Enfermedad , Células Epiteliales/patología , Técnica del Anticuerpo Fluorescente , Humanos , Inmunohistoquímica , Queratinas/análisis , Masculino , Próstata/química , Antígeno Prostático Específico/análisis , Prostatectomía , Hiperplasia Prostática/patología , Neoplasias de la Próstata/genética , Valores de Referencia , Factor de Transcripción AP-2 , Células Tumorales CultivadasRESUMEN
Smear preparations from fine-needle aspirates of 30 prostatic carcinomas obtained from radical prostatectomy specimens were examined by a dual-color fluorescence in situ hybridization (FISH) method for the presence of chromosome 7 trisomy (chromosome 9 was used as a control). The frequency of cells with trisomy 7 was determined in tumor cells and normal prostatic epithelial cells in each specimen. Comparison between the tumor and normal cells from the same patients showed that within all stages, the frequency of trisomy 7/disomy 9 cells in the tumor cells was significantly higher than that observed in the normal cells (P < 0.0001). Furthermore, the mean frequency of cells with trisomy 7/disomy 9 in advanced stages was significantly elevated over the mean frequency observed in organ-confined tumors (P = 0.02). These results are consistent with our previous data on paraffin-embedded prostate tissue sections using single-color FISH procedures. However, the method used in the present study enhances the accuracy of distinguishing trisomic 7 cells from potentially triploid (trisomy 7/trisomy 9) cells. Furthermore, the use of fine-needle aspirates rather than paraffin sections provides an easy method to examine whole nuclei. Our study also suggests that FISH provides a better measure of genetic instability (e.g., aneuploidy) in prostate tumors than flow cytometry.
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Cromosomas Humanos Par 7/genética , Trisomía , Biomarcadores , ADN de Neoplasias/análisis , Progresión de la Enfermedad , Citometría de Flujo , Humanos , Hibridación Fluorescente in Situ , Masculino , Ploidias , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/patologíaRESUMEN
We established two human prostate cancer cell lines, MDA PCa 2a and MDA PCa 2b, the TabBO model system, that reflect common features of human androgen-independent prostate cancer that are not present in other model systems: bone origin, prostate-specific antigen production, androgen receptor expression, and androgen sensitivity. We therefore hypothesized that molecular pathways in our model system reflect common alterations responsible for the progression of a subset of human prostate cancer. Progression to androgen independence has been hypothesized to be largely associated with impairment of the regulation of cell growth or apoptosis of prostate cancer cells. Therefore, in this study, we examined molecular markers known or suspected to be important in prostate cancer progression and key regulators of cell growth and apoptosis: p53, p21WAF1/CIP1, Bcl-2, Bax, retinoblastoma (Rb), and p16INK4A/MITS1. We analyzed the expression of these markers in the cell lines, their tumor of origin, and tumors derived from the cell lines by s.c. inoculation into nude mice. DNA sequencing of the entire open reading frames of the p53 and p21 genes revealed no mutations. Additionally, accumulation of the p53 protein was not found by Western blot analysis, nor was overexpression of the Bcl-2 oncoprotein detected. Bax expression was detected in MDA PCa 2a cells, whereas it was absent in MDA PCa 2b. Rb and p16 protein expression was normal as measured by both Western blot and immunochemical analyses. Immunohistochemical studies of p53, p21, Bcl-2, and Rb in both samples from the original human cancer from which the lines were derived and mouse xenografts derived from the lines revealed similar levels of protein. These results are consistent with reports indicating that 40-50% of bone metastases of prostate cancer have wild-type p53, 50-70% do not overexpress the Bcl-2 protein, and mutations in the p21 gene are rare. Therefore, we conclude that MDA PCa 2a and MDA PCa 2b reflect molecular pathways in a common subset of human androgen-independent prostate cancer and that important molecular players in apoptosis (namely, p53 and Bcl-2) seem to be intact in this subset of androgen-independent prostate cancer. Understanding the signal-transduction pathways operating in these cell lines may help to identify therapeutic targets for prostate cancer.
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Células Tumorales Cultivadas , Andrógenos/farmacología , Animales , Western Blotting , Proteínas Portadoras/análisis , División Celular/efectos de los fármacos , Inhibidor p16 de la Quinasa Dependiente de Ciclina , Inhibidor p21 de las Quinasas Dependientes de la Ciclina , Ciclinas/análisis , Ciclinas/genética , ADN de Neoplasias/química , ADN de Neoplasias/genética , Progresión de la Enfermedad , Humanos , Inmunohistoquímica , Masculino , Ratones , Ratones Desnudos , Modelos Biológicos , Mutación , Trasplante de Neoplasias , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Proteínas Proto-Oncogénicas/análisis , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Proteína de Retinoblastoma/análisis , Análisis de Secuencia de ADN , Trasplante Heterólogo , Proteína p53 Supresora de Tumor/análisis , Proteína p53 Supresora de Tumor/genética , Proteína X Asociada a bcl-2RESUMEN
The tumor grade (Gleason score) in the biopsy and pretherapy prostate-specific antigen level do not accurately predict disease outcome of individual patients' prostate cancer. We used a rapid colorimetric in situ hybridization technique to evaluate the expression level of E-cadherin (which affects cell cohesion); matrix metalloproteinases (MMPs) types 2 and 9 (which affect invasion); and vascular endothelial growth factor/vascular permeability factor (which affects angiogenesis) in archival prostatectomy specimens from 40 patients. Intratumoral heterogeneity for gene expression (edge versus center versus perineural area) was more pronounced in advanced cancers than in those that were organ confined. Regardless of Gleason score, the highest expression level for E-cadherin was found in the center or perineural area of the tumors, whereas the highest expression levels for MMP-2 and MMP-9 were associated with the invasive edge. The relationship between advancing pathological stage and expression of all four metastasis-related genes was highly significant. Decreased expression of E-cadherin and increased expression of MMP-2, MMP-9, and vascular endothelial growth factor/vascular permeability factor were associated with the Gleason score of the tumors. Irrespective of serum prostate-specific antigen level or Gleason score, the ratio between expression of MMPs and E-cadherin at the invasive edge of tumors exhibited the strongest association with nonorgan-confined prostate cancer. These data suggest that the relative expression of metastasis-related genes in radical prostatectomy specimens can distinguish between organ-confined and advanced prostate cancers and provides the rationale for a prospective study correlating gene expression in pretherapy core biopsies with outcome.
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Cadherinas/biosíntesis , Colagenasas/biosíntesis , Factores de Crecimiento Endotelial/biosíntesis , Linfocinas/biosíntesis , Metaloproteinasa 2 de la Matriz/biosíntesis , Metaloproteinasa 9 de la Matriz/biosíntesis , Neoplasias de la Próstata/diagnóstico , Neoplasias de la Próstata/metabolismo , Biopsia , Cadherinas/genética , Colagenasas/genética , Factores de Crecimiento Endotelial/genética , Humanos , Procesamiento de Imagen Asistido por Computador , Hibridación in Situ , Modelos Logísticos , Linfocinas/genética , Masculino , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Sondas de Oligonucleótidos/metabolismo , Neoplasias de la Próstata/patología , Neoplasias de la Próstata/cirugía , Factor A de Crecimiento Endotelial Vascular , Factores de Crecimiento Endotelial VascularRESUMEN
Human prostate cancer cell lines are particularly difficult to establish, and most existing cell lines do not exhibit features commonly seen in human prostate cancer. Most available models either grow only in vivo as xenografts or are androgen insensitive and fail to express prostate-specific antigen (PSA). The lack of functionally relevant model systems of advanced prostate cancer has limited prostate cancer research and therapy development. Of 30 processed samples derived from patients with prostate cancer, we established two cell lines (MDA PCa 2a and MDA PCa 2b) that express PSA and androgen receptor, grow in vitro, and are androgen sensitive. Cells from these lines produced tumors in nude mice when injected either s. c. or orthotopically (intraprostatic). Both cell lines were established from a bone metastasis of a patient whose cancer was exhibiting androgen-independent growth. Although both were derived from two samples of the same specimen, they have different genetic features (as assessed by karyotype analysis) and different phenotypes (e.g., morphology and growth rate). It is likely that they are distinct clones isolated by the use of different culture procedures and reflect the genetic heterogeneity of the tumor. These new cell lines are the first available derived from a bone metastasis of an androgen-independent prostatic adenocarcinoma that grow both in vivo and in vitro and have retained PSA expression and androgen sensitivity. They therefore constitute important model systems to address critical questions related to the androgen-independent growth of human prostate cancer and to the complex process of bone metastasis.
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Neoplasias Óseas/patología , Neoplasias Óseas/secundario , Neoplasias de la Próstata/patología , Andrógenos/farmacología , Animales , Neoplasias Óseas/genética , Neoplasias Óseas/cirugía , Técnicas de Cultivo de Célula/métodos , División Celular/efectos de los fármacos , Humanos , Cariotipificación , Masculino , Ratones , Ratones Desnudos , Persona de Mediana Edad , Orquiectomía , Antígeno Prostático Específico/sangre , Neoplasias de la Próstata/sangre , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/cirugía , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
INTRODUCTION: Chronic allograft nephropathy is the main cause of graft loss. Although many factors are involved in its development, ischemia-reperfusion injury has received increasing attention as a risk determinant. In a previous study of syngeneic renal transplantation and ischemia, we demonstrated a protective effect of acute damage by FTY 720 and antisense oligonucleotides of ICAM-1 (Oligos). The purpose of the current study was to evaluate the impact of these agents on the development of chronic graft damage in the same model. METHODS: Lewis rat were used as donors and recipients. The harvested left kidney was kept in Collins solution for 2 hours. Recipient animals received treatment with FTY 720 or Oligos or saline. At 12 and 36 weeks after transplantation, creatinine clearance, GFR, proteinuria, and arterial blood pressure were recorded. Tissue from some animals were submitted for histological studies and quantification of mRNA TGF-beta1. RESULTS: All groups showed decreased levels of GFR and creatinine clearence, but only the untreated animals showed significant deterioration compared to the pretransplant values (0.53 +/- 0.24 versus 0.21 +/- 0.24 at 36 weeks respectively; P < .05). Proteinuria was also significant in control animals at 36 weeks. Blood pressure showed a moderate increase in all groups. Histological analysis showed that treated animals had fewer signs of chronic damage according to the Banff score. All groups displayed slight increases in TGF-beta1 without differences among them. CONCLUSIONS: In this model the use of FTY or antisense oligonucleotides of ICAM-1 were associated with less functional and morphological evidence of chronic graft damage secondary to an ischemia-reperfusion injury.
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Riñón/patología , Oligonucleótidos Antisentido/uso terapéutico , Glicoles de Propileno , Daño por Reperfusión/prevención & control , Esfingosina/análogos & derivados , Animales , Modelos Animales de Enfermedad , Clorhidrato de Fingolimod , Molécula 1 de Adhesión Intercelular/genética , Inulina/farmacocinética , Riñón/efectos de los fármacos , Pruebas de Función Renal , Masculino , Ratas , Ratas Endogámicas Lew , Daño por Reperfusión/complicacionesRESUMEN
Living donation is the best choice for kidney transplantation, obtaining long-lasting good results for the recipient. Some concern still remains regarding the donor's long-term health. Kidney biopsy was routinely performed in our donor population at the time of donation many years ago. We found the existence of morphological kidney disease in those samples, in spite of normal clinical evaluations before donation. We attempted to correlate those abnormalities with long-term clinical outcomes. Donors were at least 10 years after surgery. A medical interview, including the SF-36 Health Survey, laboratory evaluation, and ambulatory blood pressure monitoring was performed on 27 donors meeting the inclusion criteria. Two donors had died after donation from unrelated causes with no known nephropathy. Histological analysis showed abnormalities in 16 of 29 donors. We found an increased prevalence of hypertension compared to the general population. Interestingly, there was no proteinuria in the donor population, and none developed clinical nephropathy. All subjects felt emotionally rewarded with donation, stating that their lives had no limitations. Our results suggest that kidney biopsy is neither necessary nor useful prior to donation because, although many donors had morphological kidney disease, none developed clinical nephropathy in the long term.