RESUMEN
PURPOSE: To develop a novel Taiwanese prostate cancer (PCa) risk model for predicting PCa, comparing its predictive performance with that of two well-established PCa risk calculator apps. METHODS: 1545 men undergoing prostate biopsies in a Taiwanese tertiary medical center between 2012 and 2019 were identified retrospectively. A five-fold cross-validated logistic regression risk model was created to calculate the probabilities of PCa and high-grade PCa (Gleason score ⧠7), to compare those of the Rotterdam and Coral apps. Discrimination was analyzed using the area under the receiver operator characteristic curve (AUC). Calibration was graphically evaluated with the goodness-of-fit test. Decision-curve analysis was performed for clinical utility. At different risk thresholds to biopsy, the proportion of biopsies saved versus low- and high-grade PCa missed were presented. RESULTS: Overall, 278/1309 (21.2%) patients were diagnosed with PCa, and 181 out of 278 (65.1%) patients had high-grade PCa. Both our model and the Rotterdam app demonstrated better discriminative ability than the Coral app for detection of PCa (AUC: 0.795 vs 0.792 vs 0.697, DeLong's method: P < 0.001) and high-grade PCa (AUC: 0.869 vs 0.873 vs 0.767, P < 0.001). Using a ≥ 10% risk threshold for high-grade PCa to biopsy, our model could save 67.2% of total biopsies; among these saved biopsies, only 3.4% high-grade PCa would be missed. CONCLUSION: Our new logistic regression model, similar to the Rotterdam app, outperformed the Coral app in the prediction of PCa and high-grade PCa. Additionally, our model could save unnecessary biopsies and avoid missing clinically significant PCa in the Taiwanese population.
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Aplicaciones Móviles , Neoplasias de la Próstata/epidemiología , Medición de Riesgo/métodos , Anciano , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Estudios Retrospectivos , TaiwánRESUMEN
BACKGROUND: Mobile health apps have emerged as useful tools for patients and clinicians alike, sharing health information or assisting in clinical decision-making. Prostate cancer (PCa) risk calculator mobile apps have been introduced to assess risks of PCa and high-grade PCa (Gleason score ≥7). The Rotterdam Prostate Cancer Risk Calculator and Coral-Prostate Cancer Nomogram Calculator apps were developed from the 2 most-studied PCa risk calculators, the European Randomized Study of Screening for Prostate Cancer (ERSPC) and the North American Prostate Cancer Prevention Trial (PCPT) risk calculators, respectively. A systematic review has indicated that the Rotterdam and Coral apps perform best during the prebiopsy stage. However, the epidemiology of PCa varies among different populations, and therefore, the applicability of these apps in a Taiwanese population needs to be evaluated. This study is the first to validate the PCa risk calculator apps with both biopsy and prostatectomy cohorts in Taiwan. OBJECTIVE: The study's objective is to validate the PCa risk calculator apps using a Taiwanese cohort of patients. Additionally, we aim to utilize postprostatectomy pathology outcomes to assess the accuracy of both apps with regard to high-grade PCa. METHODS: All male patients who had undergone transrectal ultrasound prostate biopsies in a single Taiwanese tertiary medical center from 2012 to 2018 were identified retrospectively. The probabilities of PCa and high-grade PCa were calculated utilizing the Rotterdam and Coral apps, and compared with biopsy and prostatectomy results. Calibration was graphically evaluated with the Hosmer-Lemeshow goodness-of-fit test. Discrimination was analyzed utilizing the area under the receiver operating characteristic curve (AUC). Decision curve analysis was performed for clinical utility. RESULTS: Of 1134 patients, 246 (21.7%) were diagnosed with PCa; of these 246 patients, 155 (63%) had high-grade PCa, according to the biopsy results. After confirmation with prostatectomy pathological outcomes, 47.2% (25/53) of patients were upgraded to high-grade PCa, and 1.2% (1/84) of patients were downgraded to low-grade PCa. Only the Rotterdam app demonstrated good calibration for detecting high-grade PCa in the biopsy cohort. The discriminative ability for both PCa (AUC: 0.779 vs 0.687; DeLong's method: P<.001) and high-grade PCa (AUC: 0.862 vs 0.758; P<.001) was significantly better for the Rotterdam app. In the prostatectomy cohort, there was no significant difference between both apps (AUC: 0.857 vs 0.777; P=.128). CONCLUSIONS: The Rotterdam and Coral apps can be applied to the Taiwanese cohort with accuracy. The Rotterdam app outperformed the Coral app in the prediction of PCa and high-grade PCa. Despite the small size of the prostatectomy cohort, both apps, to some extent, demonstrated the predictive capacity for true high-grade PCa, confirmed by the whole prostate specimen. Following our external validation, the Rotterdam app might be a good alternative to help detect PCa and high-grade PCa for Taiwanese men.
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Aplicaciones Móviles/normas , Neoplasias de la Próstata/diagnóstico , Medición de Riesgo/métodos , Anciano , Estudios de Cohortes , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , TaiwánRESUMEN
Protriptyline, a tricyclic anti-depressant, is used primarily to treat the combination of symptoms of anxiety and depression. However, the effect of protriptyline on prostate caner is unknown. This study examined whether the anti-depressant protriptyline altered Ca(2+) movement and cell viability in PC3 human prostate cancer cells. The Ca(2+)-sensitive fluorescent dye fura-2 was used to measure [Ca(2+)](i). Protriptyline evoked [Ca(2+)](i) rises concentration-dependently. The response was reduced by removing extracellular Ca(2+). Protriptyline-evoked Ca(2+) entry was inhibited by store-operated channel inhibitors (nifedipine, econazole and SKF96365), protein kinase C activator (phorbol 12-myristate 13 acetate, PMA) and protein kinase C inhibitor (GF109203X). Treatment with the endoplasmic reticulum Ca(2+) pump inhibitor 2,5-di-tert-butylhydr-oquinone (BHQ) in Ca(2+)-free medium inhibited 60% of protriptyline-evoked [Ca(2+)](i) rises. Conversely, treatment with protriptyline abolished BHQ-evoked [Ca(2+)](i) rises. Inhibition of phospholipase C with U73122 suppressed 50% of protriptyline-evoked [Ca(2+)](i) rises. At concentrations of 50-70 µM, protriptyline decreased cell viability in a concentration-dependent manner; which were not reversed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Collectively, in PC3 cells, protriptyline evoked [Ca(2+)](i) rises by inducing phospholipase C-associated Ca(2+) release from the endoplasmic reticulum and other stores, and Ca(2+) influx via protein kinase C-sensitive store-operated Ca(2+) channels. Protriptyline caused cell death that was independent of [Ca(2+)](i) rises.
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Apoptosis/efectos de los fármacos , Señalización del Calcio/efectos de los fármacos , Calcio/metabolismo , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Protriptilina/administración & dosificación , Transporte Biológico Activo/efectos de los fármacos , Línea Celular Tumoral , Relación Dosis-Respuesta a Droga , Humanos , Masculino , Neoplasias de la Próstata/tratamiento farmacológicoRESUMEN
OBJECTIVE: To present our experience with single mini-incision complete urinary tract exenteration (CUTE) for female dialysis patients suffering from urothelial carcinoma (UC). PATIENTS AND METHODS: Institutional review board approval was obtained. From 2005 through 2012, 14 female dialysis patients with UC underwent single mini-incision CUTE, in combination with radical hysterectomy and bilateral salpingo-oophorectomy. All were placed in the modified dorsal lithotomy position without repositioning. An infraumbilical midline mini-incision was made. Bilateral nephroureterectomy was first performed entirely extraperitoneally, followed by radical cystectomy with removal of the uterus and ovaries transperitoneally. RESULTS: All procedures were done successfully without major complications. The median operative time was 242.5 minutes, and estimated blood loss was 500 mL. The median time to oral intake was 2 postoperative days; the median hospital stay was 11 days. Ten patients remained cancer-free at a median follow-up of 46.5 months; six patients were confirmed as having preoperatively undetectable UC or renal cell carcinoma, even after reviewing preoperative computed tomography. CONCLUSIONS: This modified technique provides a time-saving complete urinary tract extirpation to eliminate preoperatively undetectable malignancy, reduce metachronous recurrences, and avert perioperative complications associated with pneumoperitoneum and repositioning. Good cancer control and early convalescence can mutually be achieved in experienced hands.
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Carcinoma de Células Transicionales/prevención & control , Diálisis , Fallo Renal Crónico/terapia , Neoplasias Renales/prevención & control , Procedimientos Quirúrgicos Mínimamente Invasivos/métodos , Exenteración Pélvica/métodos , Sistema Urinario/cirugía , Anciano , Anciano de 80 o más Años , Carcinoma de Células Transicionales/etiología , Femenino , Humanos , Fallo Renal Crónico/complicaciones , Neoplasias Renales/diagnóstico , Neoplasias Renales/etiología , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del Tratamiento , Procedimientos Quirúrgicos Urológicos/métodosRESUMEN
The aim of this study was to examine the feasibility of ureteroscope-assisted double-J stenting following laparoscopic ureterolithotomy and to evaluate the effects of retrograde ureteroscopic access exerted on the sutured ureterotomy site. From January 2002 to December 2011, 30 patients with proximal ureteral stone underwent ureteroscopic double-J stenting of the ureter following retroperitoneal laparoscopic ureterolithotomy. Patient demographics and perioperative parameters, including the degree of hydronephrosis, urine leakage, and drainage time, were retrospectively reviewed. These data were compared with those of 30 consecutive patients who received open ureterolithotomy and intracorporeal ureteral double-J stenting. In addition, a PubMed search was conducted and the related literature on the placement of a ureteral stent was reviewed. Twenty-eight patients successfully underwent ureteral double-J stenting with ureteroscopic access. No malposition of the ureteral stent was identified in the ureteroscopic group, but two patients in the intracorporeal group required postoperative adjustment of the stent. Residual stone fragments were found during stent placement in three patients in the ureteroscopic group and holmium:yttrium-aluminum-garnet laser lithotripsy was immediately performed. There was no significant difference in postoperative outcomes or complication rates between the two groups. Ureteroscope-assisted ureteral double-J stenting is a simple and safe alternative allowing intraluminal navigation along the entire ureter, correct stent placement, and prompt treatment of residual stone fragments, without radiation exposure. In addition, ureteral disruption and urinary extravasation may not be concerns for ureteroscopic access with continuous normal saline irrigation.
Asunto(s)
Laparoscopía , Stents , Uréter/cirugía , Cálculos Ureterales/cirugía , Ureteroscopios , Adulto , Anciano , Anciano de 80 o más Años , Demografía , Remoción de Dispositivos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Stents/efectos adversos , Resultado del Tratamiento , Incontinencia Urinaria/etiologíaRESUMEN
M-3M3FBS (2,4,6-trimethyl-N-(meta-3-trifluoromethyl-phenyl)-benzenesulfonamide is a presumed phospholipase C activator which induced Ca²âº movement and apoptosis in different cell models. How- ever, the effect of m-3M3FBS on cytosolic free Ca²âº concentrations ([Ca²âº]i) and apoptosis in SCM1 human gastric cancer cells is unclear. This study explored whether m-3M3FBS elevated basal [Ca²âº]i levels in suspended cells by using fura-2 as a Ca²âº-sensitive fluorescent dye. M-3M3FBS at concentrations between 5-50 µM increased [Ca²âº]i in a concentration-dependent manner. The Ca²âº signal was reduced by half by removing extracellular Ca²âº. M-3M3FBS-induced Ca²âº influx was inhibited by nifedipine, econazole, SK&F96365, aristolochic acid, and GF109203X. In Ca²âº-free medium, 50 µM m-3M3FBS pretreatment inhibited the [Ca²âº]i rise induced by the endoplasmic reticulum Ca²âº pump inhibitor thapsigargin. Conversely, pretreatment with thapsigargin partly reduced m-3M3FBS-induced [Ca²âº]i rise. Suppression of inositol 1,4,5-trisphosphate production with U73122 did not change m-3M3FBS- induced [Ca²âº]i rise. At concentrations between 25 and 50 µM m-3M3FBS killed cells in a concentration- dependent manner. The cytotoxic effect of m-3M3FBS was not reversed by prechelating cytosolic Ca²âº with acetoxy-methyl ester of bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA/AM). Annexin V/propidium iodide staining data suggest that m-3M3FBS induced apoptosis at 25 and 50 µM. M-3M3FBS also increased levels of superoxide. Together, in human gastric cancer cells, m-3M3FBS induced a [Ca²âº]i rise by inducing phospholipase C-independent Ca²âº release from the endoplasmic reticulum and Ca²âº entry via protein kinase C-sensitive store-operated Ca²âº channels. M-3M3FBS induced cell death that might involve apoptosis via reactive oxygen species production.
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Apoptosis/efectos de los fármacos , Calcio/metabolismo , Neoplasias Gástricas/tratamiento farmacológico , Sulfonamidas/farmacología , Fosfolipasas de Tipo C/fisiología , Línea Celular Tumoral , Humanos , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologíaRESUMEN
The effect of 2,4,6-trimethyl-N-(meta-3-trifluoromethyl-phenyl)-benzenesulfonamide (m-3M3FBS), a presumed phospholipase C activator, on cytosolic free Ca² ⺠concentrations ([Ca² ⺠]i ) in HA59T human hepatoma cells is unclear. This study explored whether m-3M3FBS elevated basal [Ca² ⺠]i levels in suspended cells by using fura-2 as a Ca² ⺠-sensitive fluorescent dye. M-3M3FBS at concentrations of 10- 50 µM increased [Ca² ⺠]i in a concentration-dependent fashion. The Ca² ⺠signal was reduced partly by removing extracellular Ca² ⺠. M-3M3FBS-induced Ca² ⺠influx was inhibited by nifedipine, econazole, SK&F96365, aristolochic acid, and GF109203X. In Ca² ⺠-free medium, 50 µM m-3M3FBS pretreatment inhibited the [Ca² ⺠]i rise induced by the endoplasmic reticulum Ca² ⺠pump inhibitor thapsigargin. Conversely, pretreatment with thapsigargin partly reduced m-3M3FBS-induced [Ca² ⺠]i rise. Inhibition of inositol 1,4,5-trisphosphate formation with U73122 did not alter m-3M3FBS-induced [Ca² ⺠]i rise. At concentrations between 10 and 40 µM m-3M3FBS killed cells in a concentration-dependent manner. The cytotoxic effect of m-3M3FBS was not reversed by prechelating cytosolic Ca² ⺠with 1,2-bis(2- aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Annexin V/propidium iodide staining data suggest that m-3M3FBS induced apoptosis in a concentration-dependent manner. M-3M3FBS also increased levels of reactive oxygen species. Together, in human hepatoma cells, m-3M3FBS induced a [Ca² ⺠]i rise by inducing phospholipase C-independent Ca² ⺠release from the endoplasmic reticulum and Ca² ⺠entry via protein kinase C-sensitive store-operated Ca² ⺠channels. M-3M3FBS induced cell death that might involve apoptosis via mitochondrial pathways.
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Señalización del Calcio/efectos de los fármacos , Calcio/metabolismo , Sulfonamidas/farmacología , Fosfolipasas de Tipo C/metabolismo , Apoptosis/efectos de los fármacos , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Humanos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
INTRODUCTION: There are limited data concerning the relationship between the sexual functioning of each partner in a heterosexual couple. AIM: This cross-sectional study was to investigate the association between female sexual function and the male partners' erectile function. METHODS: Two self-administered questionnaires were used, one distributed to 2,159 female employees of two hospitals in Southern Taiwan and the other to their male partners, if available, to assess sexual function in each partner of the couple. OUTCOME MEASURE: Female sexual function and male erectile function were assessed by the Female Sexual Function Index (FSFI) and by the International Index of Erectile Function (IIEF), respectively. RESULTS: Among the 1,580 female and 779 male respondents, 632 sexually active couples were eligible for the analysis with mean ages of 36.9 years (range 21-67) and 39.5 years (range 18-80) for the women and men, respectively. After adjustment for female age group, nearly all the FSFI and IIEF domain scores correlated significantly to a slight to moderate degree. On the basis of the FSFI and IIEF scores, 42.9% (255/594) of the women reported sexual difficulty, and 15.0% (96/632) of the men reported mild to moderate erectile dysfunction (ED). After adjustment for female age group, the female partners of men with ED had significantly lower total and domain scores of the FSFI than those of men without ED, with effect sizes of η(p)(2) = 0.02-0.08. After further adjustment for other risk factors, ED of the male partner was still a significant risk factor for female sexual difficulty as well as for sexual difficulty in the aspects of arousal, orgasm, sexual satisfaction, and sexual pain (odds ratio = 2.5-3.3). CONCLUSIONS: Significant correlations between female sexual functioning and male erectile function were identified.
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Disfunción Eréctil/diagnóstico , Disfunción Eréctil/psicología , Disfunciones Sexuales Psicológicas/diagnóstico , Disfunciones Sexuales Psicológicas/psicología , Esposos/psicología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Estudios Transversales , Disfunción Eréctil/epidemiología , Femenino , Encuestas Epidemiológicas , Humanos , Masculino , Matrimonio , Cómputos Matemáticos , Persona de Mediana Edad , Psicometría/estadística & datos numéricos , Disfunciones Sexuales Psicológicas/epidemiología , Estadística como Asunto , Encuestas y Cuestionarios , Taiwán , Adulto JovenRESUMEN
The effect of the natural product diindolylmethane on cytosolic Ca(2+) concentrations ([Ca(2+)](i)) and viability in PC3 human prostate cancer cells was explored. The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)](i). Diindolylmethane at concentrations of 20-50 µM induced [Ca(2+)](i) rise in a concentration-dependent manner. The response was reduced partly by removing Ca(2+). Diindolylmethane-evoked Ca(2+) entry was suppressed by nifedipine, econazole, SK&F96365, protein kinase C modulators and aristolochic acid. In the absence of extracellular Ca(2+), incubation with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) inhibited or abolished diindolylmethane-induced [Ca(2+)](i) rise. Incubation with diindolylmethane also inhibited thapsigargin or BHQ-induced [Ca(2+)](i) rise. Inhibition of phospholipase C with U73122 reduced diindolylmethane-induced [Ca(2+)](i) rise. At concentrations of 50-100 µM, diindolylmethane killed cells in a concentration-dependent manner. This cytotoxic effect was not altered by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Annexin V/PI staining data implicate that diindolylmethane (50 and 100 µM) induced apoptosis in a concentration-dependent manner. In conclusion, diindolylmethane induced a [Ca(2+)](i) rise in PC3 cells by evoking phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via phospholipase A(2)-sensitive store-operated Ca(2+) channels. Diindolylmethane caused cell death in which apoptosis may participate.
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Señalización del Calcio/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Homeostasis/efectos de los fármacos , Neoplasias de la Próstata/metabolismo , Calcio/metabolismo , Línea Celular Tumoral , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/metabolismo , Fura-2/farmacología , Humanos , Indoles/farmacología , Masculino , Tapsigargina/farmacología , Fosfolipasas de Tipo C/metabolismoRESUMEN
The effect of the natural product 3,3'-diindolylmethane (DIM) on cytosolic Ca(2+) concentrations ([Ca(2+)](i)) and viability in MG63 human osteosarcoma cells was explored. The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)](i). DIM at concentrations of 40-80 µM induced a [Ca(2+)](i) rise in a concentration-dependent manner. The response was reduced partly by removing Ca(2+). DIM-evoked Ca(2+) entry was suppressed by nifedipine, econazole, SK&F96365 and protein kinase C modulators. In the absence of extracellular Ca(2+), incubation with the endoplasmic reticulum Ca(2+) pump inhibitors thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) inhibited or abolished DIM-induced [Ca(2+)](i) rise. Incubation with DIM also inhibited thapsigargin or BHQ-induced [Ca(2+)](i) rise. Inhibition of phospholipase C with U73122 abolished DIM-induced [Ca(2+)](i) rise. At concentrations of 10-50 µM, DIM killed cells in a concentration-dependent manner. This cytotoxic effect was not altered by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Annexin V/propidium iodide staining data implicate that DIM (20 and 40 µM) induced apoptosis in a concentration-dependent manner. In sum, in MG63 cells, DIM induced a [Ca(2+)](i) rise by evoking phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via protein kinase C-sensitive store-operated Ca(2+) channels. DIM caused cell death that may involve apoptosis.
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Anticarcinógenos/farmacología , Calcio/metabolismo , Indoles/farmacología , Osteosarcoma/tratamiento farmacológico , Anticarcinógenos/administración & dosificación , Apoptosis/efectos de los fármacos , Canales de Calcio/metabolismo , Línea Celular Tumoral , Supervivencia Celular , Relación Dosis-Respuesta a Droga , Retículo Endoplásmico/metabolismo , Colorantes Fluorescentes/metabolismo , Fura-2/metabolismo , Homeostasis , Humanos , Indoles/administración & dosificación , Osteosarcoma/patología , Fosfolipasas de Tipo C/metabolismoRESUMEN
Celecoxib has been shown to have an antitumor effect in previous studies, but the mechanisms are unclear. Ca(2+) is a key second messenger in most cells. The effect of celecoxib on cytosolic free Ca(2+) concentrations ([Ca(2+)](i)) in human suspended PC3 prostate cancer cells was explored by using fura-2 as a fluorescent dye. Celecoxib at concentrations between 5 and 30 µM increased [Ca(2+)](i) in a concentration-dependent manner. The Ca(2+) signal was reduced partly by removing extracellular Ca(2+). Celecoxib-induced Ca(2+) influx was not blocked by L-type Ca(2+) entry inhibitors or protein kinase C/A modulators [phorbol 12-myristate 13-acetate (PMA), GF109203X, H-89], but was inhibited by the phospholipase A(2) inhibitor, aristolochic acid. In Ca(2+)-free medium, 30 µM of celecoxib failed to induce a [Ca(2+)](i) rise after pretreatment with thapsigargin (an endoplasmic reticulum [ER] Ca(2+) pump inhibitor). Conversely, pretreatment with celecoxib inhibited thapsigargin-induced Ca(2+) release. Inhibition of phospholipase C with U73122 did not change celecoxib-induced [Ca(2+)](i) rises. Celecoxib induced slight cell death in a concentration-dependent manner, which was enhanced by chelating cytosolic Ca(2+) with BAPTA. Collectively, in PC3 cells, celecoxib induced [Ca(2+)](i) rises by causing phospholipase C-independent Ca(2+) release from the ER and Ca(2+) influx via non-L-type, phospholipase A(2)-regulated Ca(2+) channels. These data may contribute to the understanding of the effect of celecoxib on prostate cancer cells.
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Calcio/metabolismo , Inhibidores de la Ciclooxigenasa 2/farmacología , Neoplasias de la Próstata/tratamiento farmacológico , Pirazoles/farmacología , Sulfonamidas/farmacología , Canales de Calcio Tipo L/metabolismo , Celecoxib , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Inhibidores de la Ciclooxigenasa 2/administración & dosificación , Relación Dosis-Respuesta a Droga , Retículo Endoplásmico/metabolismo , Colorantes Fluorescentes/química , Fura-2/química , Humanos , Masculino , Fosfolipasas A2/metabolismo , Neoplasias de la Próstata/patología , Pirazoles/administración & dosificación , Sulfonamidas/administración & dosificaciónRESUMEN
The effect of the antidepressant paroxetine on cytosolic free Ca2+ concentrations ([Ca2+]i) in OC2 human oral cancer cells is unclear. This study explored whether paroxetine changed basal [Ca2+]i levels in suspended OC2 cells by using fura-2 as a Ca2+-sensitive fluorescent dye. Paroxetine at concentrations between 100-1,000 microM increased [Ca2+]i in a concentration-dependent manner. The Ca2+ signal was reduced by 50% by removing extracellular Ca2+. Paroxetine-induced Ca2+ influx was inhibited by the store-operated Ca2+ channel blockers nifedipine, econazole and SK&F96365, and protein kinase C modulators. In Ca2+-free medium, pretreatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin abolished paroxetine-induced [Ca2+]i rise. Inhibition of phospholipase C with U73122 did not alter paroxetine-induced [Ca2+]i rise. Paroxetine at 10-50 microM induced cell death in a concentration-dependent manner. The death was not reversed when cytosolic Ca2+ was chelated with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. Propidium iodide staining suggests that apoptosis plays a role in the death. Collectively, in OC2 cells, paroxetine induced [Ca2+]i rise by causing phospholipase C-independent Ca2+ release from the endoplasmic reticulum and Ca2+ influx via store-operated Ca2+ channels in a manner regulated by protein kinase C and phospholipase A2. Paroxetine (up to 50 microM) induced cell death in a Ca2+-independent manner.
Asunto(s)
Apoptosis/efectos de los fármacos , Calcio/metabolismo , Neoplasias de la Boca/metabolismo , Paroxetina/farmacología , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Línea Celular Tumoral , Estrenos/farmacología , Humanos , Neoplasias de la Boca/patología , Fosfolipasas A2/fisiología , Proteína Quinasa C/fisiología , Pirrolidinonas/farmacologíaRESUMEN
The effect of the natural essential oil thymol on cytosolic Ca(2+) concentrations ([Ca(2+)](i)) and viability in human glioblastoma cells was examined. The Ca(2+)-sensitive fluorescent dye fura-2 was applied to measure [Ca(2+)](i). Thymol at concentrations of 400-1000 µM induced a [Ca(2+)](i) rise in a concentration-dependent fashion. The response was decreased partially by removal of extracellular Ca(2+). Thymol-induced Ca(2+) signal was not altered by nifedipine, econazole, SK&F96365, and protein kinase C activator phorbol myristate acetate (PMA), but was inhibited by the protein kinase C inhibitor GF109203X. When extracellular Ca(2+) was removed, incubation with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) abolished thymol-induced [Ca(2+)](i) rise. Incubation with thymol also abolished thapsigargin or BHQ-induced [Ca(2+)](i) rise. Inhibition of phospholipase C with U73122 abolished thymol-induced [Ca(2+)](i) rise. At concentrations of 200-800 µM, thymol killed cells in a concentration-dependent manner. This cytotoxic effect was not changed by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxy methyl (BAPTA/AM). Annexin V/propidium iodide staining data suggest that thymol (200, 400 and 600 µM) induced apoptosis in a concentration-dependent manner. Collectively, in human glioblastoma cells, thymol induced a [Ca(2+)](i) rise by inducing phospholipase C- and protein kinase C-dependent Ca(2+) release from the endoplasmic reticulum and Ca(2+) entry via non store-operated Ca(2+) channels. Thymol induced cell death that may involve apoptosis.
Asunto(s)
Calcio/metabolismo , Supervivencia Celular/efectos de los fármacos , Glioblastoma/patología , Homeostasis/efectos de los fármacos , Timol/farmacología , Apoptosis/efectos de los fármacos , Astrocitos/efectos de los fármacos , Astrocitos/metabolismo , Astrocitos/patología , Línea Celular Tumoral , Relación Dosis-Respuesta a Droga , Fura-2/metabolismo , Humanos , Manganeso/metabolismo , Fosfolipasas de Tipo C/metabolismoRESUMEN
The effect of the environmental contaminant, bisphenol A, on cytosolic free Ca(2+) concentrations ([Ca(2+)](i)) in Madin-Darby canine kidney (MDCK) cells is unclear. This study explored whether bisphenol A changed basal [Ca(2+)](i) levels in suspended MDCK cells by using fura-2 as a Ca(2+)-sensitive fluorescent dye. Bisphenol A, at concentrations between 50 and 300 µM, increased [Ca(2+)](i) in a concentration-dependent manner. The Ca(2+) signal was reduced, partly, by removing extracellular Ca(2+). Bisphenol A induced Mn(2+) influx, leading to quenching of fura-2 fluorescence, suggesting Ca(2+) influx. This Ca(2+) influx was inhibited by phospholipase A2 inhibitor aristolochic acid, store-operated Ca(2+) channel blockers nifedipine and SK&F96365, and protein kinase C inhibitor GF109203X. In Ca(2+)-free medium, pretreatment with the mitochondrial uncoupler, carbonylcyanide m-chlorophenylhydrazone (CCCP), and the endoplasmic reticulum Ca(2+) pump inhibitors, thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ), inhibited bisphenol A-induced Ca(2+) release. Conversely, pretreatment with bisphenol A abolished thapsigargin (or BHQ)- and CCCP-induced [Ca(2+)](i) rise. Inhibition of phospholipase C with U73122 abolished bisphenol-induced [Ca(2+)](i) rise. Bisphenol A caused a concentration-dependent decrease in cell viability via apoptosis in a Ca(2+)-independent manner. Collectively, in MDCK cells, bisphenol A induced [Ca(2+)](i) rises by causing phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and mitochondria and Ca(2+) influx via phospholipase A2-, protein kinase C-sensitive, store-operated Ca(2+) channels.
Asunto(s)
Señalización del Calcio/efectos de los fármacos , Calcio/metabolismo , Disruptores Endocrinos/toxicidad , Túbulos Renales/efectos de los fármacos , Fenoles/toxicidad , Animales , Compuestos de Bencidrilo , Técnicas de Cultivo de Célula , Línea Celular , Supervivencia Celular/efectos de los fármacos , Interpretación Estadística de Datos , Diploidia , Perros , Relación Dosis-Respuesta a Droga , Citometría de Flujo , Túbulos Renales/citología , Túbulos Renales/enzimología , Túbulos Renales/metabolismo , Inhibidores de Fosfolipasa A2 , Fosfolipasas de Tipo C/antagonistas & inhibidoresRESUMEN
Effect of the carcinogen thapsigargin on human prostate cancer cells is unclear. This study examined if thapsigargin altered basal [Ca²âº](i) levels in suspended PC3 human prostate cancer cells by using fura-2 as a Ca²âº-sensitive fluorescent probe. Thapsigargin at concentrations between 10 nM and 10 µM increased [Ca²âº](i) in a concentration-dependent fashion. The Ca²âº signal was reduced partly by removing extracellular Ca²âº indicating that Ca²âº entry and release both contributed to the [Ca²âº](i) rise. This Ca²âº influx was inhibited by suppression of phospholipase A2, but not by inhibition of store-operated Ca²âº channels or by modulation of protein kinase C activity. In Ca²âº-free medium, pretreatment with the endoplasmic reticulum Ca²âº pump inhibitor 2,5-di-(t-butyl)-1,4-hydroquinone (BHQ) nearly abolished thapsigargin-induced Ca²âº release. Conversely, pretreatment with thapsigargin greatly reduced BHQ-induced [Ca²âº](i) rise, suggesting that thapsigargin released Ca²âº from the endoplasmic reticulum. Inhibition of phospholipase C did not change thapsigargin-induced [Ca²âº](i) rise. At concentrations of 1-10 µM, thapsigargin induced cell death that was partly reversed by chelation of Ca²âº with BAPTA/AM. Annexin V/propidium iodide staining data suggest that apoptosis was partly responsible for thapsigargin-induced cell death. Together, in PC3 human prostate cancer cells, thapsigargin induced [Ca²âº](i) rises by causing phospholipase C-independent Ca²âº release from the endoplasmic reticulum and Ca²âº influx via phospholipase A2-sensitive Ca²âº channels. Thapsigargin also induced cell death via Ca²âº-dependent pathways and Ca²âº-independent apoptotic pathways.
Asunto(s)
Señalización del Calcio/efectos de los fármacos , Calcio/metabolismo , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Tapsigargina/farmacología , Apoptosis/efectos de los fármacos , Ácidos Aristolóquicos/farmacología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Estrenos/farmacología , Fluorescencia , Fura-2/metabolismo , Humanos , Espacio Intracelular/efectos de los fármacos , Espacio Intracelular/metabolismo , Masculino , Manganeso/metabolismo , Neoplasias de la Próstata/enzimología , Pirrolidinonas/farmacología , Fosfolipasas de Tipo C/metabolismoRESUMEN
The effect of sertraline, an antidepressant, on cytosolic-free Ca(2+) levels ([Ca(2+) ](i) ) in human cancer cells is unclear. This study examined whether sertraline altered basal [Ca(2+) ](i) levels in suspended PC3 human prostate cancer cells by using fura-2 as a Ca(2+) -sensitive fluorescent probe. At concentrations of 10-150 µM, sertraline induced a [Ca(2+) ](i) rise in a concentration-dependent fashion. The Ca(2+) signal was reduced partly by removing extracellular Ca(2+) indicating that Ca(2+) entry and release both contributed to the [Ca(2+) ](i) rise. Sertraline induced Mn(2+) influx, leading to quench of fura-2 fluorescence suggesting Ca(2+) influx. This Ca(2+) influx was inhibited by the suppression of store-operated Ca(2+) channels or by the modulation of protein kinase C activity. In Ca(2+) -free medium, pre-treatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin or 2,5-di-(t-butyl)-1,4-hydroquinone nearly abolished sertraline-induced Ca(2+) release. Conversely, pre-treatment with sertraline greatly reduced the inhibitor-induced [Ca(2+) ](i) rise, suggesting that sertraline released Ca(2+) from the endoplasmic reticulum. Inhibition of phospholipase C inhibited sertraline-induced [Ca(2+) ](i) rise. At 20-30 µM, sertraline killed cells in a concentration-dependent manner. The cytotoxic effect of sertraline was enhanced by chelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/AM. Annexin V-FITC data suggest that sertraline (20 and 30 µM) evoked apoptosis in a concentration-dependent manner. Together, in PC3 human prostate cancer cells, sertraline induced [Ca(2+) ](i) rises by causing phospholipase C-dependent Ca(2+) release from the endoplasmic reticulum and via multiple Ca(2+) influx pathways that involve store-operated Ca(2+) channels. Sertraline also induced apoptosis that was not triggered by [Ca(2+) ](i) rise.
Asunto(s)
Antidepresivos/farmacología , Calcio/metabolismo , Neoplasias de la Próstata/metabolismo , Sertralina/farmacología , Apoptosis , Señalización del Calcio/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Humanos , Masculino , Manganeso/metabolismo , Neoplasias de la Próstata/patología , Fosfolipasas de Tipo C/fisiologíaRESUMEN
Effect of sertraline, an antidepressant, on cytosolic free Ca(2+) levels ([Ca(2+)](i)) in human cancer cells is unclear. This study examined if sertraline altered basal [Ca(2+)](i) levels in suspended OC2 human oral cancer by using fura-2 as a Ca(2+)-sensitive fluorescent probe. At concentrations of 10-100 µM, sertraline induced a [Ca(2+)](i) rise in a concentration-dependent fashion. The Ca(2+) signal was reduced partly by removing extracellular Ca(2+) indicating that Ca(2+) entry and release both contributed to the [Ca(2+)](i) rise. Sertraline induced Mn(2+) influx, leading to quench of fura-2 fluorescence suggesting Ca(2+) influx. This Ca(2+) influx was inhibited by suppression of phospholipase A2, inhibition of store-operated Ca(2+) channels or by modulation of protein kinase C activity. In Ca(2+)-free medium, pretreatment with the endoplasmic reticulum Ca(2+) pump inhibitor thapsigargin or 2,5-di-(t-butyl)-1,4-hydroquinone (BHQ) nearly abolished sertraline-induced Ca(2+) release. Conversely, pretreatment with sertraline greatly reduced the inhibitor-induced [Ca(2+)](i) rise, suggesting that sertraline released Ca(2+) from the endoplasmic reticulum. Inhibition of phospholipase C did not change sertraline-induced [Ca(2+)](i) rise. Together, in human oral cancer cells, sertraline induced [Ca(2+)](i) rises by causing phospholipase C-independent Ca(2+) release from the endoplasmic reticulum and Ca(2+) influx via store-operated Ca(2+) channels.
Asunto(s)
Antidepresivos/farmacología , Calcio/metabolismo , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Sertralina/farmacología , Bloqueadores de los Canales de Calcio/farmacología , Línea Celular Tumoral , Inhibidores Enzimáticos/farmacología , Fluorescencia , Fura-2/metabolismo , Humanos , Manganeso/metabolismoRESUMEN
The effect of diallyl disulfide (DADS) on cytosolic Ca(2+) concentrations ([Ca(2+)](i)) and viability in PC3 human prostate cancer cells is unclear. This study explored whether DADS changed [Ca(2+)](i) in PC3 cells by using fura-2. DADS at 50-1000 µM increased [Ca(2+)](i) in a concentration-dependent manner. The signal was reduced by removing Ca(2+). DADS-induced Ca(2+) influx was not inhibited by nifedipine, econazole, SK&F96365, and protein kinase C modulators; but was inhibited by aristolochic acid. In Ca(2+)-free medium, pretreatment with the endoplasmic reticulum Ca(2+) pump inhibitors thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) nearly abolished DADS-induced [Ca(2+)](i) rise. Incubation with DADS inhibited thapsigargin or BHQ-induced [Ca(2+)](i) rise. Inhibition of phospholipase C with U73122 did not alter DADS-induced [Ca(2+)](i) rise. At 500-1000 µM, DADS killed cells in a concentration-dependent manner. The cytotoxic effect of DADS was partly reversed by prechelating cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Propidium iodide staining suggests that DADS (500 µM) induced apoptosis in a Ca(2+)-independent manner. Annexin V/PI staining further shows that 10 µM and 500 µM DADS both evoked apoptosis. DADS also increased reactive oxygen species (ROS) production. Collectively, in PC3 cells, DADS induced [Ca(2+)](i) rise probably by causing phospholipase C-independent Ca(2+) release from the endoplasmic reticulum and Ca(2+) influx via phospholipase A(2)-sensitive channels. DADS induced Ca(2+)-dependent cell death, ROS production, and Ca(2+)-independent apoptosis.
Asunto(s)
Compuestos Alílicos/farmacología , Antineoplásicos/farmacología , Señalización del Calcio/efectos de los fármacos , Disulfuros/farmacología , Neoplasias de la Próstata/tratamiento farmacológico , Apoptosis/efectos de los fármacos , Calcio/química , Calcio/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Citosol/efectos de los fármacos , Citosol/metabolismo , Ensayos de Selección de Medicamentos Antitumorales , Fura-2/química , Humanos , Masculino , Neoplasias de la Próstata/patología , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The effect of 2,4,6-trimethyl-N-(meta-3-trifluoromethyl-phenyl)-benzenesulfonamide (m-3M3FBS), a presumed phospholipase C activator, on cytosolic free Ca²âº concentrations ([Ca²âº]i) in PC3 human prostate cancer cells is unclear. This study explored whether m-3M3FBS changed basal [Ca²âº]i levels in suspended PC3 cells by using fura-2 as a Ca²âº-sensitive fluorescent dye. M-3M3FBS at concentrations between 10-50 microM increased [Ca²âº]i in a concentration-dependent manner. The Ca²âº signal was reduced by 60% by removing extracellular Ca²âº. M-3M3FBS-induced Ca²âº influx was inhibited by the store-operated Ca²âº channel blockers nifedipine, econazole and SK&F96365, and by the phospholipase A2 inhibitor aristolochic acid. In Ca²âº-free medium, 30 microM m-3M3FBS pretreatment greatly inhibited the [Ca²âº]i rise induced by the endoplasmic reticulum Ca²âº pump inhibitor thapsigargin or BHQ. Conversely, pretreatment with thapsigargin, BHQ or cyclopiazonic acid reduced the major part of m-3M3FBS-induced [Ca²âº]i rise. Inhibition of phospholipase C with U73122 did not much alter m-3M3FBS-induced [Ca²âº]i rise. Collectively, in PC3 cells, m-3M3FBS induced [Ca²âº]i rises by causing phospholipase C-independent Ca²âº release from the endoplasmic reticulum and Ca²âº influx via store-operated Ca²âº channels.