RESUMEN
Acute myelogenous leukaemia (AML) originates and is maintained by leukaemic stem cells (LSCs) that are inherently resistant to antiproliferative therapies, indicating that a critical strategy for overcoming chemoresistance in AML therapy is to eradicate LSCs. In this work, we investigated the anti-AML activity of bortezomib (BTZ), emphasizing its anti-LSC potential, using KG-1a cells, an AML cell line with stem-like properties. BTZ presented potent cytotoxicity to both solid and haematological malignancy cells and reduced the stem-like features of KG-1a cells, as observed by the reduction in CD34- and CD123-positive cells. A reduction in NF-κB p65 nuclear staining was observed in BTZ-treated KG-1a cells, in addition to upregulation of the NF-κB inhibitor gene NFΚBIB. BTZ-induced DNA fragmentation, nuclear condensation, cell shrinkage and loss of transmembrane mitochondrial potential along with an increase in active caspase-3 and cleaved PARP-(Asp 214) level in KG-1a cells. Furthermore, BTZ-induced cell death was partially prevented by pretreatment with the pancaspase inhibitor Z-VAD-(OMe)-FMK, indicating that BTZ induces caspase-mediated apoptosis. BTZ also increased mitochondrial superoxide levels in KG-1a cells, and BTZ-induced apoptosis was partially prevented by pretreatment with the antioxidant N-acetylcysteine, indicating that BTZ induces oxidative stress-mediated apoptosis in KG-1a cells. At a dosage of 0.1 mg/kg every other day for 2 weeks, BTZ significantly reduced the percentage of hCD45-positive cells in the bone marrow and peripheral blood of NSG mice engrafted with KG-1a cells with tolerable toxicity. Taken together, these data indicate that the anti-LSC potential of BTZ appears to be an important strategy for AML treatment.
Asunto(s)
Bortezomib , Leucemia Mieloide Aguda , FN-kappa B , Células Madre Neoplásicas , Estrés Oxidativo , Bortezomib/farmacología , Estrés Oxidativo/efectos de los fármacos , Humanos , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/patología , Leucemia Mieloide Aguda/metabolismo , Animales , FN-kappa B/metabolismo , Línea Celular Tumoral , Ratones , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Apoptosis/efectos de los fármacos , Antineoplásicos/farmacología , Ensayos Antitumor por Modelo de Xenoinjerto , Ratones SCIDRESUMEN
OBJECTIVE: This study seeks to investigate immunohistochemical parameters that could distinguish non-aggressive Central giant cell granuloma (CGCG) from aggressive CGCG, two groups of lesions which differ in their clinical and radiographic features and prognosis. MATERIAL AND METHODS: 12 cases of non-aggressive CGCG and 11 cases of aggressive CGCG were investigated and associated the immunohistochemical expression of macrophages (CD68 and CD163), blood vessels (CD34 and CD105), lymphatic vessels (D2-40) and regulator proteins (p63 and Ki-67). Clinical and radiographic features were also studied. RESULTS: Associations between all proteins in non-aggressive and aggressive CGCG were not significant (p > 0.05). With respect to non-aggressive CGCG, there were no significant correlations, while in aggressive CGCG there was a significant positive correlation between CD68 and CD163 (p = 0.031), between CD34 and D2-40 proteins (p = 0.04), whereas a significant negative correlation was observed between CD105 and CD68 (p = 0.040). However, regardless of aggressiveness of CGCG, there was a significant positive correlation between CD68 and CD163 (p = 0,04). Among the clinical and immunohistochemical aspects, only the symptomatology was a significant risk factor for the occurrence of aggressive CGCG (OR = 12.00/p = 0.016). CONCLUSION: Macrophages and angiogenesis contribute to their maintenance and development of CGCG. In addition, immunohistochemistry used here was not able to differentiate their aggressiveness. However, symptomatology was proved to be a risk factor for the occurrence of aggressive CGCG. It is possible that clinical features, particularly symptomatology, represent the most appropriate parameter to attempt to distinguish GCCG.
Asunto(s)
Granuloma de Células Gigantes/patología , Enfermedades Maxilomandibulares/patología , Macrófagos/patología , Neovascularización Patológica/patología , Adulto , Biomarcadores/análisis , Vasos Sanguíneos/patología , Femenino , Granuloma de Células Gigantes/metabolismo , Humanos , Inmunohistoquímica , Vasos Linfáticos/patología , Masculino , Persona de Mediana EdadRESUMEN
(1) Background: Activation of the PI3K-AKT pathway controls most hallmarks of cancer, and the hedgehog (HH) pathway has been associated with oral squamous cell carcinoma (OSCC) development and progression. We hypothesized that fibroblast-derived insulin-like growth factor-1 (IGF-1) acts in oral squamous cell carcinoma (OSCC) cells, leading to the non-canonical activation of the HH pathway, maintaining AKT activity and promoting tumor aggressiveness. (2) Methods: Primary fibroblasts (MF1) were genetically engineered for IGF-1 overexpression (MF1-IGF1) and CRISPR/Cas9-mediated IGF1R silencing was performed in SCC-4 cells. SCC-4 cells were co-cultured with fibroblasts or incubated with fibroblast conditioned medium (CM) or rIGF-1 for functional assays and the evaluation of AKT and HH pathways. (3) Results: Gene expression analysis confirmed IGF-1 overexpression in MF1-IGF1 and the absence of IGF-1 expression in SCC-4, while elevated IGF1R expression was detected. IGF1R silencing was associated with decreased survival of SCC-4 cells. Ihh was expressed in both MF1 and MF1-IGF1, and increased levels of GLI1 mRNA were observed in SCC-4 after stimulation with CM-MF1. Activation of both PI3K-AKT and the HH pathway (GLI1, Ihh and SMO) were identified in SCC-4 cells cultured in the presence of MF1-IGF1-CM. rIGF-1 promoted tumor cell proliferation, migration, invasion and tumorsphere formation, whereas CM-MF1 significantly stimulated angiogenesis. (4) Conclusions: IGF-1 exerts pro-tumorigenic effects by stimulating SCC-4 cell proliferation, migration, invasion and stemness. AKT and HH pathways were activated by IGF-1 in SCC-4, reinforcing its influence on the regulation of these signaling pathways.
Asunto(s)
Proteínas Hedgehog/metabolismo , Factor I del Crecimiento Similar a la Insulina/farmacología , Neoplasias de la Boca/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello/metabolismo , Carcinoma de Células Escamosas/patología , Técnicas de Cultivo de Célula , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Humanos , Factor I del Crecimiento Similar a la Insulina/metabolismo , Masculino , Neoplasias de la Boca/tratamiento farmacológico , Neoplasias de la Boca/patología , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Neovascularización Patológica/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Transducción de Señal/efectos de los fármacos , Carcinoma de Células Escamosas de Cabeza y Cuello/tratamiento farmacológico , Carcinoma de Células Escamosas de Cabeza y Cuello/patologíaRESUMEN
BACKGROUND: New blood vessel formation events are essential for tumor clonal expansion and progression. Despite the importance of vascular endothelial growth factor A (VEGFA) for tumor angiogenesis, few studies have evaluated the expression profile of this gene in oral squamous cell carcinoma (OSCC) and tumor margins (TM). This study aimed to evaluate the expression of the VEGFA gene and its protein in OSCC and TM. METHODS: Gene expression was evaluated in cryopreserved samples of OSCCs (n = 44), TM (n = 7), and normal mucosa from healthy patients (n = 3; NM). Quantitative PCRs were performed using the TaqMan system for the VEGFA gene, and gene expression was determined using the 2(-∆∆CQ) method. For immunohistochemical evaluation, 27 OSCC samples were embedded in a tissue microarray (TMA) block and reactions were performed using the Advance system. RESULTS: VEGFA transcript levels were 1.7-fold higher in OSCC than in NM samples. VEGFA mRNA was overexpressed in TM samples compared to NM (3.4-fold) and OSCC (2.0-fold) samples. VEGFA transcript level was overexpressed in T3-T4 tumors, metastatic lymph nodes, and stage III-IV OSCCs. Immunoreactivity of the VEGFA protein was detected in the cytoplasm of parenchymal and stromal cells, mainly in endothelial cells and fibroblasts. CONCLUSION: Our results show that VEGFA was overexpressed in aggressive OSCCs and that VEGFA expression may be an important prognostic factor in this type of cancer. Finally, tumor margins may be involved in the production of angiogenic molecules.
Asunto(s)
Carcinoma de Células Escamosas/metabolismo , Neoplasias de Cabeza y Cuello/metabolismo , Neoplasias de la Boca/metabolismo , ARN Mensajero/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Biomarcadores de Tumor/biosíntesis , Biomarcadores de Tumor/genética , Carcinoma de Células Escamosas/irrigación sanguínea , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Estudios de Casos y Controles , Citoplasma/metabolismo , Progresión de la Enfermedad , Expresión Génica , Neoplasias de Cabeza y Cuello/irrigación sanguínea , Neoplasias de Cabeza y Cuello/genética , Neoplasias de Cabeza y Cuello/patología , Humanos , Inmunohistoquímica , Neoplasias de la Boca/irrigación sanguínea , Neoplasias de la Boca/genética , Neoplasias de la Boca/patología , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Carcinoma de Células Escamosas de Cabeza y Cuello , Análisis de Matrices Tisulares , Factor A de Crecimiento Endotelial Vascular/biosíntesisRESUMEN
The canalis sinuosus (CS) is a neurovascular canal, a branch of the infraorbital canal through which the anterior superior alveolar nerve passes. There are no studies or case reports of anatomical variations related to this canal. A rare case of anatomical variation in the CS is reported that was detected by cone beam computed tomography done in a 47-year-old female as a pre-operative workup before dental implants. In this case, in the region slightly medial to tooth 23, a wide accessory branch from the CS was observed, running an intraosseous course in the inferior and posterior direction up to a foramen located in the hard palate, slightly medial in relation to tooth 23. The location of this branching, as well as its neurovascular component, is important for dental implant planning because of its proximity to the upper teeth. Identification of neurovascular bundles is fundamental to avoid complications for the patient.
Asunto(s)
Variación Anatómica , Tomografía Computarizada de Haz Cónico/métodos , Maxilar/anatomía & histología , Nervio Maxilar/anatomía & histología , Implantes Dentales , Femenino , Humanos , Maxilar/diagnóstico por imagen , Nervio Maxilar/diagnóstico por imagen , Persona de Mediana Edad , Paladar Duro/anatomía & histología , Paladar Duro/diagnóstico por imagen , Cuidados Preoperatorios/métodos , Enfermedades RarasRESUMEN
BACKGROUND: Sonic hedgehog (SHH) pathway activation has been identified as a key factor in the development of many types of tumors, including odontogenic tumors. Our study examined the expression of genes in the SHH pathway to characterize their roles in the pathogenesis of keratocystic odontogenic tumors (KOT) and ameloblastomas (AB). METHODS: We quantified the expression of SHH, SMO, PTCH1, SUFU, GLI1, CCND1, and BCL2 genes by qPCR in a total of 23 KOT, 11 AB, and three non-neoplastic oral mucosa (NNM). We also measured the expression of proteins related to this pathway (CCND1 and BCL2) by immunohistochemistry. RESULTS: We observed overexpression of SMO, PTCH1, GLI1, and CCND1 genes in both KOT (23/23) and AB (11/11). However, we did not detect expression of the SHH gene in 21/23 KOT and 10/11 AB tumors. Low levels of the SUFU gene were expressed in KOT (P = 0.0199) and AB (P = 0.0127) relative to the NNM. Recurrent KOT exhibited high levels of SMO (P = 0.035), PTCH1 (P = 0.048), CCND1 (P = 0.048), and BCL2 (P = 0.045) transcripts. Using immunolabeling of CCND1, we observed no statistical difference between primary and recurrent KOT (P = 0.8815), sporadic and NBCCS-KOT (P = 0.7688), and unicystic and solid AB (P = 0.7521). CONCLUSIONS: Overexpression of upstream (PTCH1 and SMO) and downstream (GLI1, CCND1 and BCL2) genes in the SHH pathway leads to the constitutive activation of this pathway in KOT and AB and may suggest a mechanism for the development of these types of tumors.
Asunto(s)
Ameloblastoma/genética , Perfilación de la Expresión Génica , Proteínas Hedgehog/genética , Tumores Odontogénicos/genética , Transcripción Genética/genética , Adolescente , Adulto , Ameloblastoma/química , Ameloblastos/patología , Ciclina D1/análisis , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Proteínas Hedgehog/análisis , Humanos , Masculino , Neoplasias Mandibulares/química , Persona de Mediana Edad , Mucosa Bucal/química , Recurrencia Local de Neoplasia/química , Recurrencia Local de Neoplasia/patología , Tumores Odontogénicos/química , Receptores Patched , Receptor Patched-1 , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Receptores de Superficie Celular/análisis , Receptores Acoplados a Proteínas G/análisis , Proteínas Represoras/análisis , Receptor Smoothened , Factores de Transcripción/análisis , Adulto Joven , Proteína con Dedos de Zinc GLI1RESUMEN
Liver cancers, including hepatocellular carcinoma (HCC), are the sixth most common cancer and the third leading cause of cancer-related death worldwide, representing a global public health problem. This study evaluated nine patients with HCC. Six of the cases involved hepatic explants, and three involved hepatic segmentectomy for tumor resection. Eight out of nine tumors were HCC, with one being a combined hepatocellular-cholangiocarcinoma tumor. Conventional markers of hepatocellular differentiation (Hep Par-1, arginase, pCEA, and glutamine synthetase) were positive in all patients, while markers of hepatic precursor cells (CK19, CK7, EpCAM, and CD56) were negative in most patients, and when positive, they were detected in small, isolated foci. Based on in silico analysis of HCC tumors from The Cancer Genome Atlas database, we found that Hedgehog (HH) pathway components (GLI1, GLI2, GLI3 and GAS1) have high connectivity values (module membership > 0.7) and are strongly correlated with each other and with other genes in biologically relevant modules for HCC. We further validated this finding by analyzing the gene expression of HH components (PTCH1, GLI1, GLI2 and GLI3) in our samples through qPCR, as well as by immunohistochemical analysis. Additionally, we conducted a chemosensitivity analysis using primary HCC cultures treated with a panel of 18 drugs that affect the HH pathway and/or HCC. Most HCC samples were sensitive to sunitinib. Our results offer a comprehensive view of the molecular landscape of HCC, highlighting the significance of the HH pathway and providing insight into focused treatments for HCC.
Asunto(s)
Carcinoma Hepatocelular , Proteínas Hedgehog , Neoplasias Hepáticas , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Proteínas Hedgehog/metabolismo , Proteínas Hedgehog/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Femenino , Masculino , Persona de Mediana Edad , Anciano , Regulación Neoplásica de la Expresión Génica , Biomarcadores de Tumor/metabolismo , Biomarcadores de Tumor/genética , Proteína con Dedos de Zinc GLI1/metabolismo , Proteína con Dedos de Zinc GLI1/genética , Transducción de Señal , Sunitinib/farmacología , Sunitinib/uso terapéutico , Adulto , Proteína Gli2 con Dedos de Zinc/metabolismo , Proteína Gli2 con Dedos de Zinc/genéticaRESUMEN
The purpose of this study was to evaluate the expression of Hedgehog (HH) signaling molecules (SHH and GLI-1) by cancer-associated fibroblasts (CAF) in oral squamous cell carcinoma (OSCC). Immunohistochemistry was used to detect molecular HH signaling and CAF-related protein expression, including α-SMA and S100A4, in 70 samples of human OSCC. The colocalization of α-SMA and S100A4 with SHH was also evaluated by double-staining. In vitro study was performed using primary normal oral fibroblast (NOF) and CAF through immunofluorescence and Western Blot for CAF-proteins, SHH, and GLI-1. Forty-five cases (64.28%) were positive for α-SMA exclusively in tumor stroma, and S100A4 was identified in the cytoplasm of CAFs in 94.28% (n = 66) of the cases. With respect to stromal cells, 64 (91.43%) OSCC cases were positive for SHH, and 31 were positive for GLI-1 (44.29%); positive correlations were found between SHH and α-SMA (p < 0.0001, φ = 0.51), as well as between SHH and S100A4 (p = 0.087, φ = 0.94). Protein expression of SHH and GLI-1 was observed in primary CAFs and NOFs. Although SHH was found to be localized in the cellular cytoplasm of both cell types, GLI-1 was present only in the nuclei of CAF. Our results indicate that CAFs are not only potential sources of HH ligands in tumor stroma, but may also respond to HH signaling through nuclear GLI-1 activation. We further observed that elevated SHH expression by OSCC cells was associated with higher CAF density, reinforcing the chemoattractant role played by these molecules.
Asunto(s)
Carcinoma de Células Escamosas/metabolismo , Proteínas Hedgehog/metabolismo , Neoplasias de la Boca/metabolismo , Transducción de Señal , Biomarcadores , Fibroblastos Asociados al Cáncer/metabolismo , Fibroblastos Asociados al Cáncer/patología , Carcinoma de Células Escamosas/etiología , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Técnica del Anticuerpo Fluorescente , Regulación Neoplásica de la Expresión Génica , Proteínas Hedgehog/genética , Humanos , Inmunohistoquímica , Ligandos , Neoplasias de la Boca/etiología , Neoplasias de la Boca/patología , Unión Proteica , Transporte de Proteínas , Células del Estroma/metabolismo , Células del Estroma/patología , Proteína con Dedos de Zinc GLI1/genética , Proteína con Dedos de Zinc GLI1/metabolismoRESUMEN
The aim of this investigation was to evaluate the effect of infrared (λ 846±20nm) LED irradiation on the expression profile of the extracellular matrix protein components, tenascin and fibronectin on skin wounds induced in well nourished and malnourished rats. Eighteen albino rats (21 days old) were randomly divided into a well-nourished group (standard diet) and a malnourished group (regional basic diet). After receiving the diet for 70 days, skin wounds were created and the animals were subdivided into three groups: well-nourished control (n=6), malnourished control (n=6), and malnourished+LED irradiated (λ 846±20nm, 100mW, 4J/cm(2)) (n=6). The animals were sacrificed 3 and 7 days after injury and histological sections were immunostained for both proteins. They were examined for the presence, intensity, distribution and pattern of immunolabeling. At 3 days, the distribution of tenascin was shown to be greater in the wound bed of malnourished animals compared to the well-nourished group. The intensity and distribution of tenascin was shown to be lower in the malnourished LED irradiated group compared to the malnourished control. There was a significant difference regarding the presence of fibronectin in the malnourished and well-nourished groups after 7 days (p=0.03). The intensity of fibronectin was slight (100%) in the irradiated group and moderate to intense in the malnourished control group. The results of the present study indicate that infrared LED irradiation modulates positively the expression of tenascin and particularly fibronectin.
Asunto(s)
Fibronectinas/metabolismo , Rayos Infrarrojos , Desnutrición/fisiopatología , Tenascina/metabolismo , Cicatrización de Heridas/efectos de la radiación , Animales , Expresión Génica/efectos de la radiación , Masculino , Ratas Wistar , Piel/patología , Piel/fisiopatología , Piel/efectos de la radiaciónRESUMEN
The nasopalatine canal is a long slender structure present in the midline of the anterior maxilla that connects the palate to the floor of the nasal cavity. The nasopalatine canal contains the nasopalatine nerve, the terminal branch of the nasopalatine artery, fibrous connective tissue, adipose tissue, and minor salivary glands. The purpose of this article was to report a case of a trifid nasopalatine canal detected by cone beam computed tomography prior to dental implant placement. A 47-year-old female patient was submitted to cone beam computed tomography. Axial and sagittal sections revealed a trifurcation of the nasopalatine canal. Each canal was separated from the other by bony septa and extended independently from the floor of the nasal cavity to the incisive foramen in the remnant of the alveolar process in the anterior region of the maxilla. Cone beam computed tomography has permitted better visualization of the details and anatomical variations of the nasopalatine canal. Detailed knowledge of variations in the shape, number and size of the nasopalatine canal is fundamental for surgical procedures, such as local anesthesia in the anterior maxillary region and placement of dental implants, in order to prevent damage to important arteries and nerves(AU)
El canal nasopalatino es una larga estructura delgada presente en la línea media del maxilar anterior que conecta el palato al suelo de la cavidad nasal. El canal nasopalatino contiene el nervio nasopalatino, la rama terminal de la arteria nasopalatina, el tejido conectivo fibroso, el tejido adiposo y las glándulas salivales menores. El propósito de este artículo es presentar el caso de un canal nasopalatino trifid detectado a través de tomografía computarizada de haz cónico anterior a la colocación de implantes dentales, en una paciente de femenino 47 años de edad. Secciones axiales y sagitales revelaron la trifurcación del canal nasopalatino. Cada canal se apartó del otro por tabiques ósea y extendida independientemente del suelo de la cavidad nasal para el agujero incisivo en el remanente del proceso alveolar en la región anterior del maxilar. La tomografía computarizada de haz cónico ha permitido una mejor visualización de los detalles y variaciones anatómicas del canal nasopalatino. El conocimiento detallado de las variaciones en su forma, el número y el tamaño del canal nasopalatino es fundamental para los procedimientos cirúrgicos, así como la anestesia local en la región anterior del maxilar superior y la colocación de los implantes dentales, con el fin de prevenir el daño a las arterias y a los nervios importantes(AU)