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Organ-confined prostate cancer of low-grade histopathology is managed with radiation, surgery, active surveillance, or watchful waiting and exhibits a 5-year overall survival (OS) of 95%, while metastatic prostate cancer (PCa) is incurable, holding a 5-year OS of 30%. Treatment options for advanced PCa-metastatic and non-metastatic-include hormone therapy that inactivates androgen receptor (AR) signaling, chemotherapy and genome-targeted therapy entailing synthetic lethality of tumor cells exhibiting aberrant DNA damage response, and immune checkpoint inhibition (ICI), which suppresses tumors with genomic microsatellite instability and/or deficient mismatch repair. Cancer genome sequencing uncovered novel somatic and germline mutations, while mechanistic studies are revealing their pathological consequences. A microRNA has shown biomarker potential for stratifying patients who may benefit from angiogenesis inhibition prior to ICI. A 22-gene expression signature may select high-risk localized PCa, which would not additionally benefit from post-radiation hormone therapy. We present an up-to-date review of the molecular and therapeutic aspects of PCa, highlight genomic alterations leading to AR upregulation and discuss AR-degrading molecules as promising anti-AR therapeutics. New biomarkers and druggable targets are shaping innovative intervention strategies against high-risk localized and metastatic PCa, including AR-independent small cell-neuroendocrine carcinoma, while presenting individualized treatment opportunities through improved design and precision targeting.
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Background Intravenous sufentanil-midazolam and inhalational sevoflurane are widely used for anesthetic induction in children undergoing cardiac surgery. However, knowledge about their effects on hemodynamics and cardiac efficiency remains limited due largely to the lack of direct monitoring method. We used minimally invasive technique pressure recording analytical method (PRAM) to directly monitor hemodynamics and cardiac efficiency and compared the effects of the two anesthetic regimens in children undergoing ventricular septal defect repair. Methods Forty-Four children (2.3±0.9 years) were randomly divided into two groups to receive either intravenous sufentanil (1 µg/kg) and midazolam (0.2 mg/kg) (Group SM) or 2.0 minimal alveolar concentration (MAC) sevoflurane (Group S) to complete induction after sedation was obtained with 2.0 MAC sevoflurane. Systemic hemodynamic data recorded by PRAM included heart rate (HR), systolic (SBP) and mean (MBP) blood pressure, stroke volume index (SVI), cardiac index (CI), systemic vascular resistance index (SVRI), the maximal slope of systolic upstroke (dp/dtmax) and cardiac cycle efficiency (CCE) after sedation obtained, 1, 2, 5 min after induction achieved, 1, 2, 5 and 10 min after intubation. Results HR, SVRI showed a decrease in Group SM but an increase in Group S (Ptime*group<0.0001) in the study period. SVI and CCE showed an increase in Group SM but a decrease in Group S (Ptime*group<0.0001). SBP, MBP and CI were related to time after polynomial transformation, and showed an increase after intubation in Group SM but a decrease in Group S (Ptime2*group<0.0001). Conclusion PRAM provides meaningful and direct monitoring of hemodynamics and cardiac efficiency during the dynamic period of anesthetic induction in children undergoing cardiac surgery. As compared to inhalational sevoflurane, intravenous sufentanil-midazolam exerts more favorable effects on systemic hemodynamics and cardiac efficiency during anesthetic induction in this group of patients.
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Anestesia Geral/métodos , Comunicação Interventricular/cirurgia , Hemodinâmica/fisiologia , Monitorização Intraoperatória/métodos , Sevoflurano/administração & dosagem , Sufentanil/administração & dosagem , Analgésicos Opioides , Anestésicos Inalatórios/administração & dosagem , Pré-Escolar , Feminino , Seguimentos , Comunicação Interventricular/fisiopatologia , Humanos , Injeções Intramusculares , Masculino , Estudos Prospectivos , Fatores de TempoRESUMO
BACKGROUND: An emerging paradigm holds that resistance to the development of allergic diseases, including allergic rhinoconjunctivitis, relates to an intact epithelial/epidermal barrier during early childhood. Conceivably, the immunologic and genomic footprint of this resistance is preserved in nonatopic, nonallergic adults and is unmasked during exposure to an aeroallergen. OBJECTIVE: The aim of this study was to obtain direct support of the epithelial/epidermal barrier model for allergic rhinoconjunctivitis. METHODS: Twenty-three adults allergic to house dust mites (HDMs) (M+) and 15 nonsensitive, nonallergic (M-) participants completed 3-hour exposures to aerosolized HDM (Dermatophagoides pteronyssinus) powder on 4 consecutive days in an allergen challenge chamber. We analyzed: (1) peripheral blood leukocyte levels and immune responses; and (2) RNA sequencing-derived expression profiles of nasal cells, before and after HDM exposure. RESULTS: On HDM challenge: (1) only M+ persons developed allergic rhinoconjunctivitis symptoms; and (2) peripheral blood leukocyte levels/responses and gene expression patterns in nasal cells were largely concordant between M+ and M- participants; gross differences in these parameters were not observed at baseline (pre-exposure). Two key differences were observed. First, peripheral blood CD4+ and CD8+ T-cell activation levels initially decreased in M- participants versus increased in M+ participants. Second, in M- compared with M+ participants, genes that promoted epidermal/epithelial barrier function (eg, filament-aggregating protein [filaggrin]) versus inflammation (eg, chemokines) and innate immunity (interferon) were upregulated versus muted, respectively. CONCLUSION: An imprint of resistance to HDM challenge in nonatopic, nonallergic adults was muted T-cell activation in the peripheral blood and inflammatory response in the nasal compartment, coupled with upregulation of genes that promote epidermal/epithelial cell barrier function.
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Alérgenos/imunologia , Antígenos de Dermatophagoides/imunologia , Conjuntivite Alérgica/imunologia , Pyroglyphidae/imunologia , Rinite Alérgica/imunologia , Administração por Inalação , Adulto , Animais , Conjuntivite Alérgica/genética , Resistência à Doença , Células Epiteliais/imunologia , Células Epiteliais/metabolismo , Feminino , Proteínas Filagrinas , Humanos , Contagem de Leucócitos , Masculino , Mucosa Nasal/imunologia , Mucosa Nasal/metabolismo , Rinite Alérgica/genética , TranscriptomaRESUMO
Fluid management is challenging in infants after cardiopulmonary bypass. Pulse pressure variation (PPV) derived from pressure recording analytical method (PRAM) is based on lung-heart interaction during mechanical ventilation. A prospective observational study conducted in operating room tested PPV to predict fluid responsiveness in ventricular septal defect infants. Infants in open chest conditions with median sternotomy (n = 26) or minimally invasive right thoracotomy (n = 29) undergoing ventricular septal defect repair were enrolled. After cardiopulmonary bypass and modified ultrafiltration, all patients received fluid challenge. PPV was recorded using PRAM along with heart rate, diastolic blood pressure, stroke volume index (SVI), and cardiac index (CI) before and after volume replacement. Patients were considered as responders to fluid loading when CI increased ≥15%. In infants with median sternotomy, 12 were responders and 14 non-responders. PPV in responders was higher than that in non-responders (24.7 ± 6.4 vs. 16.6 ± 5.0%, P < 0.01). Area under the curve was 0.85 (95% confidence interval, 0.69-1, P = 0.001) and cutoff value 19% with a sensitivity of 92% and a specificity of 71%. In infants with minimally invasive right thoracotomy, 16 were responders and 13 non-responders. PPV in responders was higher than that in non-responders (25.0 ± 6.8 vs. 18.2 ± 5.3, P < 0.01). Area under the curve was 0.83 (95 confidence interval, 0.66-0.98, P = 0.001) and cutoff value 18% with a sensitivity of 94% and a specificity of 69%. PPV sensitively predicts fluid responsiveness in ventricular septal defect infants after surgical repair in open chest conditions both with median sternotomy and minimally invasive right thoracotomy.
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Determinação da Pressão Arterial/métodos , Pressão Sanguínea/fisiologia , Ponte Cardiopulmonar/métodos , Hidratação/métodos , Comunicação Interventricular/cirurgia , Área Sob a Curva , Feminino , Comunicação Interventricular/fisiopatologia , Hemodinâmica/fisiologia , Humanos , Lactente , Masculino , Procedimentos Cirúrgicos Minimamente Invasivos/métodos , Estudos Prospectivos , Curva ROC , Sensibilidade e Especificidade , Esternotomia/métodos , Toracotomia/métodosRESUMO
BACKGROUND: Sevoflurane and ketamine are commonly used to obtain sedation and facilitate intravenous anesthetic induction in children undergoing cardiac surgery who are uncooperative. We used a new and direct systemic hemodynamic monitoring technique pressure recording analytical method and compared the hemodynamic effects of sevoflurane and ketamine to facilitate intravenous anesthetic induction. METHODS: Forty-four children with ventricular septal defect (2.2â±â1.2 years) were enrolled and randomized to receive sevoflurane (Group S) or intramuscular ketamine (GroupâK) for sedation, followed by intravenous midazolam-sufentanil induction and tracheal intubation. Recorded parameters included heart rate (HR), arterial pressures, stroke volume index (SVI), cardiac index (CI), systemic vascular resistance index (SVRI), the maximal slope of systolic upstroke (dp/dtmax) after sedation obtained with sevoflurane or ketamine, 1, 2, 5âminutes after midazolam-sufentanil, 1, 2, 5, and 10âminutes after tracheal intubation. Rate-pressure product (RPP) and cardiac power output (CPO) were calculated. RESULTS: As compared with Group S, Group K had faster decreases during intravenous anesthetic induction in arterial pressures (Pâ<â.01 for all), higher HR, arterial pressures, SVRI, dp/dtmax, RPP, lower SVI, CI, CPO (Pâ<â.05 for all) during the study period. CONCLUSION: As compared with sevoflurane, ketamine facilitated intravenous anesthetic induction exerts unfavorable effects on systemic hemodynamic and myocardial energetic in children with ventricular septal defect.
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Anestésicos Dissociativos/administração & dosagem , Anestésicos Inalatórios/administração & dosagem , Comunicação Interventricular/cirurgia , Hemodinâmica/efeitos dos fármacos , Ketamina/administração & dosagem , Éteres Metílicos/administração & dosagem , Pré-Escolar , Feminino , Humanos , Lactente , Intubação Intratraqueal , Masculino , Estudos Prospectivos , Sevoflurano , Resultado do TratamentoRESUMO
BACKGROUND: Modifiers of symptom severity in patients with allergic rhinoconjunctivitis (AR) are imprecisely characterized. The hygiene hypothesis implicates childhood microbial exposure as a protective factor. Cockroach sensitization (C+) might be a proxy for microbial exposure. OBJECTIVE: We sought to determine whether C+ assayed by means of skin prick tests influenced AR symptom severity in controlled and natural settings. METHODS: Total symptom scores (TSSs) were recorded by 21 participants with house dust mite allergy (M+) in the natural setting and during repeated exposures of 3 hours per day to house dust mite allergen in an allergen challenge chamber (ACC). In M+ participants the peripheral blood and nasal cells were assayed for T-cell activation and transcriptomic profiles (by using RNA sequencing), respectively. Participants allergic to mountain cedar (n = 21), oak (n = 34), and ragweed (n = 23) recorded TSSs during separate out-of-season exposures to these pollens (any pollen sensitization [P+]) in the ACC; a subset recorded TSSs in the pollination seasons. RESULTS: The hierarchy of TSSs (highest to lowest) among M+ participants tracked the following skin prick test sensitization statuses: M+P+C- > M+P+C+ > M+P-C- > M+P-C+. In nasal cells and peripheral blood the immune/inflammatory responses were rapidly resolved in M+P+C+ compared with M+P+C- participants. Among those allergic to pollen, C+ was associated with a lower TSS during pollen challenges and the pollination season. After aggregated analysis of all 4 ACC studies, C+ status was associated with a 2.8-fold greater likelihood of a lower TSS compared with C- status (odds ratio, 2.78; 95% CI, 1.18-6.67; P = .02). CONCLUSIONS: C+ status is associated with mitigation of AR symptom severity in adults with AR.
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Alérgenos/administração & dosagem , Baratas/imunologia , Conjuntivite Alérgica/terapia , Dessensibilização Imunológica/métodos , Pólen/imunologia , Rinite Alérgica Sazonal/terapia , Adulto , Alérgenos/química , Alérgenos/imunologia , Ambrosia/química , Ambrosia/imunologia , Animais , Baratas/química , Conjuntivite Alérgica/diagnóstico , Conjuntivite Alérgica/imunologia , Conjuntivite Alérgica/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Pólen/química , Pyroglyphidae/química , Pyroglyphidae/imunologia , Rinite Alérgica Sazonal/diagnóstico , Rinite Alérgica Sazonal/imunologia , Rinite Alérgica Sazonal/fisiopatologia , Estações do Ano , Índice de Gravidade de Doença , Testes CutâneosRESUMO
OBJECTIVE: To investigate the expression and regulation of colony-stimulating factor 1 (CSF-1) and its receptor, C-FMS, in endometriosis. DESIGN: In vivo and vitro study. SETTING: University-based academic medical center. PATIENT(S): Reproductive-age women undergoing surgery for benign conditions. INTERVENTION(S): Peritoneal and endometrial tissue samples were obtained. MAIN OUTCOME MEASURE(S): CSF-1 and C-FMS expression. RESULT(S): Significantly higher CSF-1 levels were found in peritoneal fluid of patients with endometriosis compared with control subjects. Ectopic endometriotic tissue had 3.5-fold and 1.7-fold increases in CSF-1 and C-FMS expression, respectively, compared with eutopic tissue. Coculture of endometrial cells from either established cell lines or patient samples with peritoneal mesothelial cells (PMCs) led to increased expression of CSF-1 and C-FMS. A higher but nonsignificant increase in levels of C-FMS and CSF-1 was found in cocultures of endometrial epithelial cells from patients with endometriosis compared with those without endometriosis. CONCLUSION(S): Increased CSF-1 levels may contribute to endometriosis lesion formation and progression. Elevation in CSF-1 after coculture of endometrial cells with PMCs suggests that endometrial tissue may be a source of peritoneal CSF-1. Increased C-FMS expression in endometrial cells from women with endometriosis cocultured with PMCs suggests that endometrial tissue involved in lesion formation is highly responsive to CSF-1 signaling.
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Endometriose/imunologia , Endométrio/imunologia , Fator Estimulador de Colônias de Macrófagos/metabolismo , Receptor de Fator Estimulador de Colônias de Macrófagos/metabolismo , Centros Médicos Acadêmicos , Líquido Ascítico/imunologia , Comunicação Celular , Células Cultivadas , Técnicas de Cocultura , Endometriose/genética , Endometriose/patologia , Endométrio/patologia , Células Epiteliais/imunologia , Células Epiteliais/patologia , Feminino , Humanos , Fator Estimulador de Colônias de Macrófagos/genética , Peritônio/imunologia , Peritônio/patologia , Receptor de Fator Estimulador de Colônias de Macrófagos/genética , Células Estromais/imunologia , Células Estromais/patologia , Texas , Regulação para CimaRESUMO
A common single nucleotide polymorphism (SNP) in the gene of brain-derived neurotrophic factor (BDNF) results from a substitution at position 66 from valine (Val) to methionine (Met) and may predispose to human neuropsychiatric disorders. We proposed to determine whether these BDNF gene SNPs were associated with fibromyalgia syndrome (FMS) and/or any of its typical phenotypes. Patients with FMS (N = 95) and healthy normal controls (HNC, N = 58) were studied. Serum high-sensitivity C-reactive protein (hsCRP) levels were measured using an enzyme-linked immunosorbent assay (ELISA). The BDNF SNPs were determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).The BDNF SNP distribution was 65 (68%) Val/Val, 28 (30%) Val/Met, and 2 (2%) Met/Met for FMS and 40 (69%), 17(29%), and 1 (2%) for HNC, respectively. The serum high-sensitivity C-reactive protein (hsCRP)and body mass index (BMI) in FMS were higher than in HNC. The FMS with BDNF Val66Val had significantly higher mean BMI (P = 0.0001) and hsCRP (P = 0.02) than did FMS carrying the Val66Met genotype. This pattern was not found in HNC. Phenotypic measures of subjective pain, pain threshold, depression, or insomnia did not relate to either of the BDNF SNPs in FMS. The relative distribution BDNF SNPs did not differ between FMS and HNC. The BDNF Val66Met polymorphism is not selective for FMS. The BDNF Val66Val SNP identifies a subgroup of FMS with elevated hsCRP and higher BMI. This is the first study to associate a BDNF polymorphism with a FMS subgroup phenotype.
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Índice de Massa Corporal , Fator Neurotrófico Derivado do Encéfalo/genética , Proteína C-Reativa/análise , Fibromialgia/genética , Polimorfismo de Nucleotídeo Único , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Ensaio de Imunoadsorção Enzimática , Feminino , Fibromialgia/sangue , Fibromialgia/diagnóstico , Fibromialgia/imunologia , Frequência do Gene , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Fenótipo , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Medição de Risco , Fatores de Risco , Texas , Regulação para CimaRESUMO
The development of nonhuman primate (NHP) embryonic stem cell (ESC) models holds great promise for cell-mediated treatment of debilitating diseases and to address numerous unanswered questions regarding the therapeutic efficacy of ESCs while supplanting ethical considerations involved with human studies. Here we report successful establishment and characterization of 3 novel baboon (Papio cynocephalus) ESC lines from the inner cell mass of intracytoplasmic sperm injection-derived blastocysts. Embryos were cultured in an improved baboon embryo in vitro culture protocol. The inner cell mass of blastocyst was laser-dissected and plated on mouse embryonic fibroblast feeder cell monolayer in the NHP ESC culture medium. Three cell lines with characteristic ESC morphology have been cultured through an extended period (>14 months), with 2 male cell lines (UT-1 and -2) and 1 female cell line (UT-3) displaying normal baboon karyotypes. Reverse transcription-polymerase chain reaction analysis confirmed that all 3 lines express primate ESC pluripotency markers, including OCT-4, NANOG, SOX-2, TERT, TDGF, LEFTYA, and REX-1. All 3 lines demonstrated positive immunocytochemical staining for OCT-4, stage-specific embryonic antigen-3, stage-specific embryonic antigen-4, TRA-1-60, and TRA-1-81. Baboon ESCs injected into NOD/SCID mice formed teratomas with all 3 germ layers. In addition, embryoid body-like spherical structures were derived and initial outgrowth was observed when embedded into extracellular matrix Matrigel. The ESC lines established in this NHP model have the potential to extend our knowledge in the fields of developmental biology, regenerative medicine, and future applications, including preclinical safety assessment of in vivo stem cell therapy.
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Blastocisto/citologia , Técnicas de Cultura de Células/métodos , Células-Tronco Embrionárias/citologia , Fertilização in vitro , Papio/embriologia , Animais , Blastocisto/metabolismo , Agregação Celular/genética , Diferenciação Celular/genética , Linhagem Celular , Dissecação , Corpos Embrioides/citologia , Corpos Embrioides/metabolismo , Células-Tronco Embrionárias/metabolismo , Feminino , Humanos , Imuno-Histoquímica , Cariotipagem , Masculino , Camundongos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Teratoma/patologiaRESUMO
OBJECTIVE: We previously demonstrated that adherence of endometrial epithelial (EECs) and stromal cells (ESCs) to peritoneal mesothelial cells (PMCs) is partly regulated by ESC/EEC CD44 interactions with PMC associated hyaluronan. CD44, a transmembrane glycoprotein and major ligand for hyaluronan, has numerous splice variants which may impact hyaluronan binding. Here, we assessed whether ESCs and EECs from women with endometriosis demonstrate increased adherence to PMCs and examined CD44 splice variants' potential role in this process. DESIGN: In vitro study. SETTING: Academic medical center. PATIENT(S): Fertility patients with and without endometriosis. INTERVENTION(S): Menstrual endometrium was collected from women with and without endometriosis confirmed surgically. The adherence of ESC/EECs to PMCs was measured. The ESC/EEC CD44 splice variants were assessed using dot-blot analysis. RESULT(S): The ESCs and EECs from women with endometriosis demonstrated increased adherence to PMCs. The predominant CD44 splice variants expressed by ESCs and EECs from women with and without endometriosis were v3, v6, v7, v8, v9, and v10. The ESCs and EECs from women with endometriosis were more likely to express v6, v7, v8, and v9. CONCLUSION(S): Increased eutopic endometrial-PMC adherence and CD44 splice variant expression may contribute to the histogenesis of endometriotic lesions. Elucidation of factors controlling this expression may lead to novel endometriosis therapies.
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Endometriose/patologia , Endométrio/patologia , Endométrio/fisiopatologia , Receptores de Hialuronatos/genética , Doenças Peritoneais/patologia , Peritônio/fisiologia , Sequência de Bases , Adesão Celular/fisiologia , Células Cultivadas , Epitélio/patologia , Epitélio/fisiopatologia , Feminino , Humanos , Receptores de Hialuronatos/metabolismo , Ciclo Menstrual/fisiologia , Peritônio/metabolismo , Peritônio/patologia , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Células Estromais/metabolismo , Células Estromais/patologia , Células Estromais/fisiologia , Regulação para Cima/genéticaRESUMO
A new class of platinum(II) coordination complexes and their dye tagged conjugates has been synthesized from N-substituted diaminocyclohexane ligands. The in vitro anticancer activities of the platinum compounds have been validated against the breast cancer cell-line MCF-7 and the normal cell-line MCF-10A via sulforhodamine B and colony formation assay. The platinum compounds and the corresponding metal-free ligands exhibited higher drug efficiencies than cisplatin and oxaliplatin against MCF-7 cells. Cellular uptake and DNA-bound Pt were demonstrated by atomic absorption spectroscopy. The platinum complexes displayed increased cellular accumulation and DNA binding as compared with cisplatin. Real-time reverse transcription polymerase chain reaction assay was employed to investigate drug effects on mRNA expression in MCF-7 cells. The results indicated that the study compounds are effective in regulating cyclin D1, Bcl-2, and p53 genes; yet, oxaliplatin is less effective in manipulating those genes. The luminescent probe that was integrated into the platinum complexes made it possible to monitor cellular drug distribution using optical imaging. Targeting of tumor cell nuclei by the study compounds was confirmed by confocal microscopy. Taken together, these new platinum(II)-based antitumor agents are different from marketed platinum drugs in several critical aspects and could have potential in cancer therapy.
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Antineoplásicos/toxicidade , Corantes Fluorescentes/química , Compostos Organoplatínicos/química , Antineoplásicos/síntese química , Antineoplásicos/química , Linhagem Celular Tumoral , Cisplatino/toxicidade , Ciclina D1/genética , Ciclina D1/metabolismo , DNA/metabolismo , Regulação da Expressão Gênica , Humanos , Microscopia Confocal , Compostos Organoplatínicos/síntese química , Compostos Organoplatínicos/toxicidade , Oxaliplatina , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismoRESUMO
Transforming growth factor beta 1 (TGF-beta1) levels are increased in the peritoneal fluid of endometriosis patients, and endometrial cells express TGF-beta signaling components; however, little is known regarding the role of TGF-beta in endometriosis. Our objective was to examine the effects of TGF-beta1 on (i) the expression of macrophage colony-stimulating factor receptor encoded by the c-fms gene, (ii) transmesothelial invasiveness of endometrial cells, (iii) cellular proliferation and (iv) attachment to peritoneal mesothelial cells (PMCs). Effects of TGF-beta1 on c-fms mRNA expression were determined by real-time RT-PCR and c-fms cell-surface expression by flow cytometry. Effects of TGF-beta1 on the invasiveness of the immortalized endometrial epithelial cell (EEC) line EM42 and primary EECs were examined using a three-dimensional in vitro system modeling the peritoneum. Cellular proliferation and attachment to PMCs were also examined using established techniques. TGF-beta1 had little or no effect on cellular proliferation and endometrial cell attachment to PMCs. TGF-beta1 significantly induced the expression of c-fms mRNA and c-fms cell-surface expression. TGF-beta1 enhanced transmesothelial invasion by EM42 cells and EECs. Antagonists of TGF-beta1 signaling significantly inhibited both the induction of c-fms expression and cellular invasiveness, suggesting that additional studies are warranted to assess the therapeutic potential of TGF-beta antagonists in endometriosis.
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Endométrio/citologia , Células Epiteliais/citologia , Receptor de Fator Estimulador de Colônias de Macrófagos/metabolismo , Fator de Crescimento Transformador beta/farmacologia , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Imunoprecipitação da Cromatina , Inibidores Enzimáticos/farmacologia , Células Epiteliais/efeitos dos fármacos , Feminino , Citometria de Fluxo , Expressão Gênica/efeitos dos fármacos , Humanos , Imidazóis/farmacologia , Regiões Promotoras Genéticas/genética , Piridinas/farmacologia , Receptor de Fator Estimulador de Colônias de Macrófagos/genética , Fator de Crescimento Transformador beta/antagonistas & inibidores , Fator de Crescimento Transformador beta/fisiologiaRESUMO
We report a new series of Herceptin-platinum(II) binding complexes, Her-nLPt(II) (Her denotes Herceptin; L denotes diamino ligands and L=L1-L4; n=1, 5, or 10). Solution chemistry studies have shown that these complexes are stable under physiological conditions (pH 7.4 in PBS). The platinum(II) compound L1Pt(II)Cl(2) inhibits the growth of a panel of human cancer cell lines at sub-micromolar concentrations. Remarkable cancer-cell-specific cytotoxicity was observed with Her-nL1Pt(II) (n=1, 5, 10) toward Her2/neu-overexpressing cancer cells (SK-BR-3 and SK-OV-3) over normal fibroblast cells. Annexin V apoptosis assays in SK-BR-3 and low-Her2/neu-expressing MCF-7 breast cancer cells further confirmed the critical role of Herceptin with this cancer-cell-specific agent. It was also found that the L1Pt(II)Cl(2) complex is an efficient regulator of the apoptotic genes Bcl-2 in the treated SK-BR-3 cells. Also, enhanced regulatory effects were observed in Her-10L1Pt(II). Taken together, this study suggests a new approach for the development of mAb-platinum(II)-based targeting agents for the treatment of human cancers.
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Anticorpos Monoclonais/química , Anticorpos Monoclonais/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Compostos Organoplatínicos/química , Compostos Organoplatínicos/farmacologia , Anticorpos Monoclonais Humanizados , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Conformação Molecular , Receptor ErbB-2/antagonistas & inibidores , Reprodutibilidade dos Testes , Estereoisomerismo , TrastuzumabRESUMO
A set of 12 enantiomeric diamine-based small molecules was synthesized and screened for anticancer activity against five human cancer cell lines: NCI-H460, A549, MCF-7, SK-BR-3, and T-47D. The salicyl diamino compounds (1-6) were found to induce inhibition of the growth of cancer cells at submicromolar concentrations. The lead compound, N,N'-bis-salicyl-(1R,2R)-diaminocyclohexane (1) displayed single-reagent anticancer activity with an IC(50) value equal to or less than 2.0 microM in H460 and A-549 cancer cells. SRB and colony formation assays indicated that compound 1 shows greater cytotoxic activity toward MCF-7 cells than MCF-10A cells. Real-time RT-PCR analysis demonstrated that compound 1, is an extremely efficient regulator of antiapoptotic genes, Bcl-xL, Bcl-2, and the cell cycle related gene, cyclin D1. This study provides a new insight into the development of novel small molecules in the treatment of human breast cancers.
Assuntos
Antineoplásicos/química , Diaminas/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Diaminas/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Espectroscopia de Ressonância Magnética , RNA Mensageiro/antagonistas & inibidores , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Espectrometria de Massas por Ionização por Electrospray , EstereoisomerismoRESUMO
Cervical cancer is the third most common gynecologic cancer in the United States. The presence and possible involvement of several cytokines have been studied in cervical cancer; however, very little data, if any, are available on whether cervical tumors are responsive to stimulation by the macrophage colony-stimulating factor-1 (CSF-1). Given the involvement of c-fms and its ligand CSF-1 in gynecologic cancers, such as that of the uterus and the ovaries, we have examined the expression of c-fms and CSF-1 in cervical tumor (n = 17) and normal cervix (n = 8) samples. The data show that c-fms and its ligand are significantly higher in cervical carcinomas compared with normal samples. Immunohistochemistry not only showed that tumor cells expressed significantly higher levels of c-fms but also c-fms levels were markedly higher in tumor cells than tumor-associated stromal cells. Blocking c-fms activity in cervical cancer cells, which express CSF-1 and c-fms, resulted in increased apoptosis and decreased motility compared with control, suggesting that CSF-1/c-fms signaling may be involved in enhanced survival and possibly invasion by cervical cancer cells via an autocrine mechanism. Combined, the data show for the first time the induction of CSF-1 and c-fms in cervical carcinomas and suggest that c-fms activation may play a role in cervical carcinogenesis. Additionally, our data suggest that transforming growth factor-beta1 may be a factor in inducing the expression of c-fms in cervical cancer cells. The data suggest that c-fms may be a valuable therapeutic target in cervical cancer.
Assuntos
Carcinoma/genética , Regulação Neoplásica da Expressão Gênica , Fator Estimulador de Colônias de Macrófagos/metabolismo , Receptor de Fator Estimulador de Colônias de Macrófagos/metabolismo , Fator de Crescimento Transformador beta1/fisiologia , Neoplasias do Colo do Útero/genética , Apoptose/efeitos dos fármacos , Carcinoma/metabolismo , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Feminino , Células HeLa , Humanos , Fator Estimulador de Colônias de Macrófagos/antagonistas & inibidores , Fator Estimulador de Colônias de Macrófagos/genética , Receptor de Fator Estimulador de Colônias de Macrófagos/genética , Fator de Crescimento Transformador beta1/metabolismo , Células Tumorais Cultivadas , Neoplasias do Colo do Útero/metabolismoRESUMO
Various proteins involved in transcriptional regulation possess highly selective DNA-binding domains, known as zinc fingers. However, little is known about small-molecule zinc(II) complexes in the regulation of gene expression and programmed cell death. A new family of zinc(II) complexes is reported, which might be useful against human cancer cells. By using template synthesis and in vitro cell-line screening, a set of zinc(II) complexes has been found to induce apoptosis of cancer cells and display single-reagent in vitro cytotoxicity. The method used to synthesize the molecules resulted in "built-in" luminescent behavior. Confocal optical imaging clearly demonstrated penetration through the cell membrane by these metal complexes. We have discovered that C3, the meso-zinc(II) complex is an extremely efficient regulator of the cell cycle and anti-apoptosis genes bcl-2 and bcl-xL. This study provides a new insight into the development of zinc(II) complexes as potential drugs.
Assuntos
Antineoplásicos/farmacologia , Proteínas Reguladoras de Apoptose/genética , Genes cdc/efeitos dos fármacos , Compostos Organometálicos , Zinco/química , Apoptose , Neoplasias da Mama/tratamento farmacológico , Ciclo Celular , Linhagem Celular Tumoral , Regulação para Baixo/efeitos dos fármacos , Feminino , Humanos , Concentração Inibidora 50 , Estrutura Molecular , Compostos Organometálicos/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismoRESUMO
UNLABELLED: A growing body of evidence suggests that early growth response-1 (Egr-1), a transcription factor, may function as a tumor suppressor. The aim of this study was to gain more evidence to support the role of Egr-1 in the suppression of cancer cell growth and to examine the potential correlation between Egr-1 and gelsolin. MATERIALS AND METHODS: Histochemical staining coupled with breast cancer tissue arrays were used to examine the expression levels of Egr-1 and gelsolin. Reporter assays and gel shift were used to study the transcriptional activity of Egr-1 on the regulation of gelsolin. RESULTS: Our data showed that most normal mammary tissues expressed high levels of Egr-1, while the majority of breast cancer tissues expressed very small amounts of Egr-1. The expression pattern of Egr-1 in human breast cancer tissues was highly correlated with gelsolin expression. Induction of Egr-1 by serum stimulation accompanied the increase of gelsolin expression. In cells lacking the induction of Egr-1 in response to serum stimulation, gelsolin expression remained unchanged. Furthermore, gelsolin promoter activity was profoundly reduced in Egr-1 null mouse embryonic fibroblasts compared to Egr-1 wild-type mouse embryonic fibroblasts. Gel shift experiments indicated that Egr-1 can directly bind to the gelsolin promoter. CONCLUSION: Our results suggest that Egr-1 may be an important breast cancer marker and that an as yet uncharacterized pathway involved in Egr-1 and gelsolin expression exists which leads to breast cancer cell development.
Assuntos
Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Proteína 1 de Resposta de Crescimento Precoce/genética , Proteína 1 de Resposta de Crescimento Precoce/metabolismo , Gelsolina/genética , Gelsolina/metabolismo , Regulação Neoplásica da Expressão Gênica , Animais , Sítios de Ligação , Neoplasias da Mama/genética , Linhagem Celular Tumoral , Regulação para Baixo , Humanos , Imuno-Histoquímica , Camundongos , Regiões Promotoras Genéticas/genética , Fatores de TempoRESUMO
Epidemiologic studies have implicated estrogenic exposure as well as human papilloma virus (HPV) infection in cervical carcinogenesis, and some studies have suggested that estrogen and HPV may play synergistic roles in cervical tumorigenesis. In this study, we report a novel finding that approximately 35% of cervical carcinomas tested (n = 19) express aromatase, the enzyme responsible for converting androgen to estrogen, the rate-limiting and final step in estrogen biosynthesis. On the other hand, no aromatase expression was detected in precancerous (n = 42) or normal cervical (n = 17) tissue samples. Increased aromatase was associated with increases in estrogen receptors (ER-alpha and ER-beta) and a decrease in progesterone receptor levels, suggesting that in situ estrogen signaling via ER may be involved in tumor growth. Stable overexpression of aromatase in HPV+ cervical cancer cells resulted in increased cellular proliferation, anchorage-independent growth, and ER expression and activity. In contrast, little change in ER was observed in HPV- cells. Steroid hormone receptor expression observed in vitro paralleled that seen in cervical carcinomas expressing aromatase. Aromatase overexpression also induced the expression of cyclin D1, proliferating cell nuclear antigen, and the HPV oncogenes, E6 and E7. Furthermore, the data underscores the importance of steroid receptor (estrogen and progesterone receptors) regulation in cervical carcinogenesis. To our knowledge, this is the first report demonstrating the induction of aromatase expression in cervical carcinomas, and opens the possibility that aromatase inhibitors may be potential therapeutic agents in cervical carcinomas expressing aromatase.
Assuntos
Aromatase/biossíntese , Estrogênios/metabolismo , Neoplasias do Colo do Útero/enzimologia , Aromatase/metabolismo , Processos de Crescimento Celular/fisiologia , Linhagem Celular Tumoral , Colo do Útero/enzimologia , Ciclina D1/metabolismo , Indução Enzimática , Receptor alfa de Estrogênio/biossíntese , Receptor beta de Estrogênio/biossíntese , Feminino , Humanos , Células-Tronco Neoplásicas/enzimologia , Células-Tronco Neoplásicas/patologia , Papillomaviridae , Proteínas E7 de Papillomavirus/genética , Infecções por Papillomavirus/enzimologia , Infecções por Papillomavirus/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Receptores de Progesterona/biossíntese , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/virologiaRESUMO
Aromatase transgenic mice exhibit hyperplastic and dysplastic changes, attesting to the importance of local estrogen in breast carcinogenesis. These mice also show increased levels of the estrogen receptor alpha and beta (ERalpha, ERbeta) suggesting that this receptor may play an important role in the initiation of estrogen-mediated mammary hyperplasia observed in these mice. To address the specific role of ERalpha in the mammary development and in the induction of estrogen-mediated hyperplasia in aromatase transgenic mice, we have generated MMTV-aromatase x ERalpha knockout cross (referred as aromatase/ERKO). Even though ERbeta is expressed in aromatase/ERKO mice, lack of ERalpha leads to impaired mammary growth in these mice. The data suggest that ERalpha plays an important role in the mammary gland development as well as in the induction of mammary hyperplasia in aromatase transgenic mice. Lack of ERalpha expression in the aromatase/ERKO mice resulted in a decrease in the expression of Cyclin D1, PCNA and TGFbeta relative to the aromatase parental strain. The studies involving aromatase/ERKO mice show that lack of ERalpha results in impaired mammary development even in the presence of continuous tissue estrogen, suggesting estrogen/ERalpha-mediated actions are critical for mammary development and carcinogenesis.
Assuntos
Aromatase/genética , Receptor alfa de Estrogênio/fisiologia , Glândulas Mamárias Animais/crescimento & desenvolvimento , Glândulas Mamárias Animais/patologia , Animais , Aromatase/metabolismo , Ciclina D1/metabolismo , Receptor alfa de Estrogênio/genética , Receptor alfa de Estrogênio/metabolismo , Hiperplasia/metabolismo , Hiperplasia/patologia , Masculino , Glândulas Mamárias Animais/imunologia , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Antígeno Nuclear de Célula em Proliferação/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismoRESUMO
A number of recent studies have suggested that the colony-stimulating factor (CSF-1) and its receptor c-fms may be involved in the development of mammary glands during lactation and breast cancer. To study the role of CSF-1 or its receptor in initiation of mammary tumorigenesis, we have generated two independent lines of transgenic mice that overexpress either CSF-1 or c-fms under the control of the mouse mammary tumor virus promoter. Mammary glands of the virgin CSF-1 transgenic mice show increased ductal branching, hyperplasia, dysplasia, and other preneoplastic changes, which are indicative of increased cellular proliferation. Similar changes were also evident in the mammary glands of the c-fms transgenic mice. These changes became more prominent with age and resulted in mammary tumor formation. Moreover, secondary events like dimethylbenz(a)anthracene treatment accelerated mammary tumor formation in these mice. Although the expression of estrogen receptor alpha was not significantly changed in either of the transgenic mouse strains, progesterone receptor levels was higher in both transgenic lines as compared with the nontransgenic littermates. Expression of G1 cyclins was prominently increased in the mammary glands of both the CSF-1 and c-fms transgenic lines, suggesting increased cell cycle progression in these strains. In addition, the proliferation marker proliferating cell nuclear antigen (PCNA) and the mitogen-responsive transcription factor c-jun were also increased as compared with the nontransgenic controls. These findings, along with the histological data, support the hypothesis that CSF-1 and its receptor are involved in the etiology of breast cancer.