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OBJECTIVE: This study is to explore the correlation between the contrast-enhanced ultrasound (CEUS) characteristics of breast cancer and the epithelial-mesenchyme transformation (EMT). METHODS: Totally 119 patients of breast cancer underwent CEUS. Tissues in the active area were collected and subjected to the immunohistochemical detection, PT-PCR and Western blot. Correlation analysis was conducted between the clinical pathological parameters and the CEUS indicators. RESULTS: The expression levels of CD44, N-cadherin, and ß-catenin in breast cancer tissues were higher than those in adjacent tissues (P<0.05). However, the expression levels of CD24 and E-cadherin in breast cancer tissues were lower than those in adjacent tissues (P<0.05). There was no significant difference in E-cadherin mRNA and Vimentin levels between cancer and adjacent tissues (P>0.05). The expressions were up-regulated in the CSCs, with higher histological grade, lymph node metastasis, and negative estrogen receptor (ER) expression. Smaller breast tumors, with no lymph node metastasis, lower clinical stage, and positive ER expression, tended to exhibit the up-regulated epithelial phenotype. Breast tumors, with high histological grade, lymph node metastasis, high clinical staging grade, and negative ER expression, tended to exhibit the up-regulated interstitial phenotype. The peak intensity of the time-intensity curve (TIC) for the CEUS was positively correlated with the CSC marker CD44 and the interstitial phenotype marker N-cadherin. The starting time of enhancement was negatively correlated with the N-cadherin. Area under the curve was positively correlated with the expression of CD44 and N-cadherin, while negatively correlated with the epithelial phenotype marker ß-catenin. The time to peak was negatively correlated with the interstitial phenotypes Vimentin and N-cadherin, with no correlation with the E-cadherin or ß-catenin. CONCLUSION: Breast cancers show the enlarged lesions after enlargement and perfusion defect for the CEUS. The fast-in pattern, high enhancement, and high perfusion in the TIC are correlated with the CSCs and EMT expressions, suggesting poor disease prognosis.
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Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/patología , Medios de Contraste/metabolismo , Transición Epitelial-Mesenquimal , Microcirculación , Células Madre Neoplásicas/patología , Ultrasonografía Mamaria/métodos , Neoplasias de la Mama/diagnóstico por imagen , Neoplasias de la Mama/metabolismo , Femenino , Humanos , Persona de Mediana Edad , Células Madre Neoplásicas/metabolismo , PronósticoRESUMEN
BACKGROUND: This study is to examine the feasibility of shear wave elastography (SWE) anisotropy in assessing the prognosis of breast cancer. METHODS: We enrolled 119 breast cancer patients from January 2017 to October 2019. SWE was performed before operation. Emax (maximum elasticity value), Emean (average elasticity value), Esd (standard deviation of the lesion elasticity value), Eratio (elasticity value of adipose tissue), anisotropy coefficient and difference were recorded. After operation, we collected clinical pathological data, and performed immunohistochemistry and real-time PCR tests on CD44, CD24, E-cadherin, ß-catenin, vimentin and N-cadherin. Finally, we analyzed the correlation among parameters of SWE, anisotropy and clinicopathology, and markers of CSCs (cancer stem cells) and EMT (epithelial-mesenchymal transition). RESULTS: Emax, Emean and Esd of the cross section were higher than those of the longitudinal section. Breast cancer with a higher elastic modulus was often accompanied by a hyperechoic halo, which was manifested as mixed echo and post-echo attenuation, and was accompanied by a higher BI-RADS (breast imaging reporting and data system) classification. When breast cancer had hyperechoic halo and weakened posterior echo, SWE of the lesion showed more obvious anisotropy. In addition, larger diameter of the longitudinal section indicated higher stiffness of the cross section. Correlation analysis showed that E-cadherin was negatively correlated with SWE in longitudinal section. CD44, N-cadherin, ß-catenin were positively correlated with SWE in longitudinal and cross sections. Vimentin and CD24 had no correlation with SWE parameters. CONCLUSION: SWE of breast cancer is anisotropic. The cross-sectional SWE is better than the longitudinal SWE, Emax is better than Emean, the anisotropy of SWE is better than SWE, and the anisotropy factor is better than the anisotropy difference.
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Neoplasias de la Mama/diagnóstico por imagen , Neoplasias de la Mama/patología , Diagnóstico por Imagen de Elasticidad/métodos , Transición Epitelial-Mesenquimal , Células Madre Neoplásicas , Adulto , Anciano , Anciano de 80 o más Años , Anisotropía , Biomarcadores de Tumor/metabolismo , Módulo de Elasticidad , Estudios de Factibilidad , Femenino , Humanos , Persona de Mediana Edad , PronósticoRESUMEN
This paper aimed to investigate the application of ultrasound image segmentation technology based on the back propagation neural network (BPNN) artificial intelligence algorithm in the diagnosis of breast cancer axillary lymph node metastasis, thereby providing a theoretical basis for clinical diagnosis. In this study, 90 breast cancer patients with axillary lymph node metastasis were selected as the research objects and rolled randomly into an experimental group and a control group. Besides, all of them were examined by ultrasound. The BPNN algorithm for the ultrasound image segmentation diagnosis method was applied to the patiens from the experimental group, while the control group was given routine ultrasound diagnosis. Thus, the value of this algorithm in ultrasonic diagnosis was compared and explored. The results showed that when the number of hidden layer nodes based on the BPNN artificial intelligence algorithm was 2, 3, 4, 5, 6, 7, and 8, the corresponding segmentation accuracy was 97.3%, 96.5%, 94.8%, 94.8%, and 94.1% in turn. Among them, the segmentation accuracy was the highest when the number of hidden layer nodes was 2. The correlation of independent variable bubble plot analysis showed that the presence or absence of capsules, the presence of crab feet or burrs in breast cancer lesions was critical influencing factors for the occurrence of axillary lymph node metastasis, and the standardized importance was 99.7% and 70.8%, respectively. Besides, the area under the two-dimensional receiver operating characteristic (ROC) curve of the BPNN artificial intelligence algorithm model classification was always greater than the area under the curve of manual segmentation, and the segmentation accuracy was 90.31%, 94.88%, 95.48%, 95.44%, and 97.65% in sequence. In addition, the segmentation specificity of different running times was higher than that of manual segmentation. In conclusion, the BPNN artificial intelligence algorithm had high accuracy, sensitivity, and specificity for ultrasound image segmentation, with a better segmentation effect. Therefore, it had a better diagnostic effect for breast cancer axillary lymph node metastasis.
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Neoplasias de la Mama , Algoritmos , Inteligencia Artificial , Neoplasias de la Mama/diagnóstico , Femenino , Humanos , Metástasis Linfática/diagnóstico por imagen , Sensibilidad y Especificidad , Tecnología , UltrasonografíaRESUMEN
Circular RNAs (circRNAs) are tissue-specific RNAs with a more stable structure than linear RNAs, and their association with breast cancer (BC) is poorly understood. This study examined the biological effects of circ_0000043 in the progression of BC. In this study, expression of circ_0000043 in BC tissue samples was measured using quantitative real-time polymerase chain reaction. Immunohistochemistry and Western blot were used to detect the expression of Smad family member 3 (Smad3). CCK-8, wound healing, and Transwell assays were used to assess the effect of circ_0000043 on the proliferation, migration, and invasiveness of BC cells. Moreover, the binding relationships between circ_0000043 and miR-136, and miR-136 and Smad3 were detected by dual-luciferase reporter assay. Additionally, Western blot was used to detect the expressions of markers related to epithelial-mesenchymal transition, including E-cadherin, N-cadherin, and vimentin. Our results show that the expression of circ_0000043 is up-regulated in BC tissues and cell lines. The proliferation, migration, invasiveness, and epithelial-mesenchymal transition of BC cells were significantly inhibited by knockdown of circ_0000043, and overexpression of circ_0000043 had the opposite effects. Additionally, circ_0000043 up-regulate the expression of Smad3 by sponging miR-136. In conclusion, our study demonstrates that circ_0000043 promote the progression of BC via regulating the miR-136-Smad3 axis.
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Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/patología , Transición Epitelial-Mesenquimal , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , ARN Circular/genética , Proteína smad3/metabolismo , Apoptosis , Biomarcadores de Tumor/genética , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Movimiento Celular , Proliferación Celular , Femenino , Humanos , Persona de Mediana Edad , Invasividad Neoplásica , Pronóstico , Proteína smad3/genética , Tasa de Supervivencia , Células Tumorales Cultivadas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Silicosis is a chronic fibrotic lung disease caused by the accumulation of silica dust in the distal lung. Canonical Wnt signaling and NADPH oxidase 4 (NOX4) have been demonstrated to play a crucial role in the pathogenesis of pulmonary fibrosis including silicosis. However, the underlying mechanisms of crosstalk between these two signalings are not fully understood. In the present study, we aimed to explore the interaction of Wnt/ß-catenin and NOX4 of human epithelial cells in response to an exposure of silica dust. Results demonstrated an elevated expression of key components of Wnt/ß-catenin signaling and NOX4 in the lungs of silicon dioxide- (SiO2-) induced silicosis mice. Furthermore, the activated Wnt/ß-catenin and NOX4 signaling are accompanied by an inhibition of cell proliferation, an increase of ROS production and cell apoptosis, and an upregulation of profibrogenic factors in BEAS-2B human lung epithelial cells exposed to SiO2. A mechanistic study further demonstrated that the Wnt3a-mediated activation of canonical Wnt signaling could augment the SiO2-induced NOX4 expression and reactive oxygen species (ROS) production but reduced glutathione (GSH), while Wnt inhibitor DKK1 exhibited an opposite effect to Wnt3a. Vice versa, an overexpression of NOX4 further activated SiO2-induced Wnt/ß-catenin signaling and NFE2-related factor 2 (Nrf2) antioxidant response along with a reduction of GSH, whereas the shRNA-mediated knockdown of NOX4 showed an opposite effect to NOX4 overexpression. These results imply a positive feed forward loop between Wnt/ß-catenin and NOX4 signaling that may promote epithelial-mesenchymal transition (EMT) of lung epithelial cells in response to an exposure of silica dust, which may thus provide an insight into the profibrogenic role of Wnt/ß-catenin and NOX4 crosstalk in lung epithelial cell injury and pathogenesis of silicosis.
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Células Epiteliales/metabolismo , Transición Epitelial-Mesenquimal/efectos de los fármacos , Pulmón/metabolismo , NADPH Oxidasa 4/metabolismo , Mucosa Respiratoria/metabolismo , Dióxido de Silicio/toxicidad , Vía de Señalización Wnt/efectos de los fármacos , Animales , Células Epiteliales/patología , Femenino , Pulmón/patología , Masculino , Ratones , Mucosa Respiratoria/patología , Silicosis/metabolismo , Silicosis/patologíaRESUMEN
Nonluminal breast cancer has high early metastasis and treatment resistance, and neoadjuvant chemotherapy (NAC) is needed. The presence of cancer stem cells (CSC) and epithelial-mesenchymal transition (EMT) leads to poor prognosis. This study investigated the changes in CSC markers and EMT markers after NAC in nonluminal breast cancer and their correlation with contrast-enhanced ultrasound (CEUS) features and chemotherapy efficacy. Before NAC, the range of nonluminal breast cancer on CEUS was larger than that of two-dimensional ultrasound, but after NAC, it was significantly smaller than that of two-dimensional ultrasound and closer to the postoperative pathological size. After NAC, the enlarged lesions and perfusion defects were significantly less than those before NAC. The time-intensity curve showed the characteristics of slow-in, low enhancement, and low perfusion. Nonluminal breast cancer downregulated the expression of CSC markers and EMT markers after NAC, but the epithelial phenotype of nonluminal breast cancer with good response to chemotherapy was upregulated. In nonluminal breast cancer with poor response to chemotherapy, markers of CSC and EMT were highly expressed before chemotherapy. In conclusion, CEUS is better than conventional ultrasound in estimating NAC efficacy in this mode. CEUS can also predict the prognosis of nonluminal breast cancer before NAC with the characteristics of enhanced enlargement and perfusion defects. The contrast-enhanced time-intensity curve of lesions with relatively poor blood supply may have more CSC and EMT characteristics.
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Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/diagnóstico por imagen , Neoplasias de la Mama/tratamiento farmacológico , Medios de Contraste/química , Transición Epitelial-Mesenquimal , Terapia Neoadyuvante , Células Madre Neoplásicas/metabolismo , Ultrasonografía Mamaria , Adulto , Anciano , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Persona de Mediana Edad , Células Madre Neoplásicas/patología , Perfusión , ARN Mensajero/genética , ARN Mensajero/metabolismo , Adulto JovenRESUMEN
The present study aimed to investigate the correlation between ultrasonographic features, basic fibroblast growth factor (bFGF), and the local invasiveness of papillary thyroid carcinoma (PTC).A total of 350 samples of thyroid nodules were collected. Routine ultrasonography was performed before the operation and routine pathological diagnosis and bFGF detection were performed after the operation.'These 350 samples of thyroid nodules included 90 samples of nodular goiter, 36 samples of focal thyroiditis, and 224 samples of PTC. A total of 326 thyroid nodules were examined for bFGF. The results revealed that the difference in the expression of bFGF between the benign and malignant groups was statistically significant (Pâ<â.05) and the difference in the positive expression of bFGF between the invasive and non-invasive PTC groups was statistically significant (Pâ<â.05).Whether the shape of PTC is regular or not and whether there is micro-calcification in PTC and other ultrasonographic features, the size and location of the lesions and the age of the patient help make a preliminary prediction of local invasiveness before the operation. Postoperative detection of bFGF is helpful for further risk assessments of PTC.
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Factor 2 de Crecimiento de Fibroblastos/metabolismo , Cáncer Papilar Tiroideo/diagnóstico por imagen , Cáncer Papilar Tiroideo/metabolismo , Neoplasias de la Tiroides/diagnóstico por imagen , Neoplasias de la Tiroides/metabolismo , Adulto , Factores de Edad , Anciano , Biomarcadores de Tumor/metabolismo , Femenino , Bocio/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Estudios Prospectivos , Cáncer Papilar Tiroideo/patología , Neoplasias de la Tiroides/patología , Nódulo Tiroideo/metabolismo , Tiroiditis/metabolismo , Ultrasonografía , Adulto JovenRESUMEN
BACKGROUND The aim of this study was to investigate the perfusion characteristics of different breast lesion regions in contrast-enhanced ultrasonography (CEUS). MATERIAL AND METHODS A total of 161 malignant and benign breast lesion cases were subjected to CEUS. Perfusion parameters were analyzed and compared between the central and peripheral lesion regions, and surrounding tissue. Mass section was marked with methylene blue. Samples were subjected to immunohistochemistry, and microvessel density (MVD) was calculated. RESULTS There were significant differences in perfusion performance between the central and peripheral lesion regions, and surrounding tissue. In the malignant tumors, the fast-in and fast-out pattern was the most common type in the peripheral region (57.98%), while the slow-in and slow-out patterns were the major types in the central region and surrounding tissue (49.58% and 57.98%, respectively). Compared with the surrounding tissue, the peripheral region in the cancers exhibited hyperechoic enhancement and fast-in and slow-out pattern, with large area under the curve (AUC), while the central region showed isoechoic enhancement and equally-in and slow-out pattern, with large AUC. In the benign lesions, the peripheral region exhibited hyperechoic enhancement and fast-in and fast-out pattern, with small AUC, while the central region showed isoechoic enhancement and equally-in and -out pattern, with the same AUC value. Moreover, the perfusion parameters in the central and peripheral regions were significantly associated with MVD. CONCLUSIONS It is more objective to evaluate the perfusion performance of breast lesions with the reference of surrounding tissue. Compared with the central region, the peripheral region could better reflect the perfusion characteristics of malignant lesions.
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Neoplasias de la Mama/irrigación sanguínea , Neoplasias de la Mama/diagnóstico por imagen , Ultrasonografía/métodos , Adulto , Anciano , Anciano de 80 o más Años , China , Medios de Contraste , Femenino , Humanos , Inmunohistoquímica , Microvasos/diagnóstico por imagen , Persona de Mediana Edad , Neovascularización Patológica , Coactivador 3 de Receptor Nuclear/metabolismoRESUMEN
OBJECTIVE: This study is to investigate the diagnostic role of multi-mode ultrasound in level 4 BI-RADS breast lesions and to establish a Logistic regression model. METHODS: Totally 179 patients with 182 sites of breast lesions were enrolled in this study. Preoperatively, the examinations of routine ultrasonography, elastography, contrast-enhanced ultrasonography and three-dimensional color Doppler were performed. Postoperatively, the breast lesions were diagnosed as benign and malignant lesions according to pathological results. Diagnostic indicators of each ultrasound analysis were determined and compared. The relationship between these diagnostic indicators and the benign and malignant features of breast lesions was analyzed by single factor analysis. Logistic regression model was established. RESULTS: The diagnostic indicators with high sensitivity and specificity were tumor edge, enhanced range and score of elastography. Four factors of tumor edge, enhanced order, contrast mode and score of elastography were related with the benign and malignant features of breast lesions. The prediction model was Logit (P) = 0.636 + 4.471X1 + 4.337X2 + 3.753X3 + 3.014X4 + 2.525X5 + 2.105X6. Likelihood ratio test showed that the model was statistically significant (χ(2) = 161.876, P < 0.0001). This model could effectively distinguish between benign and malignant tumors (R(2) = 0.813, prediction accuracy 92.3%). The differences in sensitivity and specificity between multi-mode ultrasound diagnosis and routine ultrasound diagnosis were statistically significant (P < 0.001). However, there was no significant difference between Logistic regression model and multi-mode ultrasound diagnosis. CONCLUSION: Multi-mode ultrasound and Logistic regression model are more effective in diagnosing level 4 BI-RADS breast lesions.
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The aim of this study was to explore the association of (99m)Tc-HYNIC-octreotide SPECT/CT somatostatin receptor scintigraphy (SRS) with the pathological grading and expression of somatostatin receptor 2 (SSTR2) for meningioma, and to define possible roles of SRS in the pathological grading of meningioma. Thirty patients with meningiomas diagnosed by MRI and treated with (99m)Tc-HYNIC-octreotide SPECT/CT SRS. Meningioma tissues were obtained from analyzing pathological grading and measuring the expression of SSTR2 with immunohistochemical staining. The meningioma side (T) to the contralateral side (NT) ratios (T/TN) of radioactive counts were calculated to investigate their association with the pathological grading of meningioma and the expression of SSTR2. All 30 cases showed high meningioma radioactivity accumulation using SRS with a sensitivity of 100 %, while CT scans only detected 25 cases with a sensitivity of 83 %. Twenty cases with grade I meningioma had a T/NT ratio of 3.80 ± 1.67, which was significantly lower than the other 10 cases (9.57 ± 3.78) with a grade II meningioma (P < 0.01). All meningiomas expressed SSTR2 as detected by immunohistochemical staining, and the T/NT ratio was positively associated with the pathological grading of meningioma and the expression of SSTR2 (with r of 0.784 and 0.805, respectively). (99m)Tc-HYNIC-octreotide SPECT/CT SRS is a sensitive technique for detecting meningioma, and the T/NT ratio of the SRS data closely correlates with the pathological grade of meningioma and the expression of SSTR2.
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Neoplasias Meníngeas/patología , Meningioma/patología , Octreótido/análogos & derivados , Compuestos de Organotecnecio , Receptores de Somatostatina/metabolismo , Tomografía Computarizada de Emisión de Fotón Único , Tomografía Computarizada por Rayos X , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Estudios de Seguimiento , Humanos , Procesamiento de Imagen Asistido por Computador , Técnicas para Inmunoenzimas , Imagen por Resonancia Magnética , Masculino , Neoplasias Meníngeas/diagnóstico por imagen , Neoplasias Meníngeas/metabolismo , Meningioma/diagnóstico por imagen , Meningioma/metabolismo , Persona de Mediana Edad , Clasificación del Tumor , Pronóstico , RadiofármacosRESUMEN
PURPOSE: The aim of this study was to determine whether recurrence of meningiomas could be reduced by combining somatostatin receptor scintigraphy (SRS) of Tc-HYNIC-octreotide SPECT/CT and radio guidance with a hand-held γ-probe during surgery. MATERIALS AND METHODS: Thirty patients with meningiomas diagnosed by MRI and considered as the study group were treated with Tc-HYNIC-octreotide SPECT/CT preoperatively and pathologically examined postoperatively. Another 60 patients considered as the control group underwent only an MRI preoperatively and a pathological examination postoperatively. For the patients in the study group, meningiomas were removed by a hand-held γ-probe 4-12 h after SRS; these patients were followed up by MRI examination each year for 5 years to monitor the recurrence rate of the meningiomas. For the control group, routine operations without radio guidance were performed and followed up with MRI examination simultaneously. RESULTS: All patients in the study group, comprising 20 with grade I and 10 with grade II meningiomas, showed high Tc-HYNIC-octreotide accumulation with a sensitivity of 100% for SRS; four patients (13.3%) relapsed after a 5-year follow-up, including one (5%) patient with a grade I and three (30%) patients with a grade II meningioma. However, among the 60 control patients, 30 were of grade I and 30 were of grade II; 18 patients (30%) experienced recurrence, including five (16.7%) grade I patients and 13 (43.3%) grade II patients. There were significant differences in recurrence between the study group and the control group when considering all the patients and those in grade I and grade II (all P values were below 0.001). CONCLUSION: Tc-HYNIC-octreotide SPECT/CT SRS is a sensitive technique for detecting meningiomas, and radio guidance using a hand-held γ-probe with Tc-HYNIC-octreotide during surgery can significantly reduce the recurrence of meningiomas.
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Meningioma/diagnóstico por imagen , Meningioma/prevención & control , Imagen Multimodal/métodos , Recurrencia Local de Neoplasia/prevención & control , Octreótido/análogos & derivados , Compuestos de Organotecnecio/farmacología , Tomografía de Emisión de Positrones , Radiocirugia/métodos , Receptores de Somatostatina/metabolismo , Tomografía Computarizada por Rayos X , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Femenino , Rayos gamma , Humanos , Imagen por Resonancia Magnética/métodos , Masculino , Meningioma/terapia , Persona de Mediana Edad , Octreótido/farmacología , Factores de TiempoRESUMEN
OBJECTIVE: The goal of this study was to detect the intertumoral heterogeneity of CT10, CT45 and GAGE7 expression and further to analyze their prognostic value. METHODS: The intertumoral heterogeneity of three cancer/testis antigens was examined by immunohistochemistry using 120 samples from patients with infiltrating ductal breast carcinoma. The expression patterns were classified and correlated with the clinicopathologic variables and outcome of the patients. RESULTS: CT10 showed punctate, focal and diffuse expression patterns according to the characteristic of its distribution. CT45 showed cytoplasmic, nuclear or combined cytoplasmic and nuclear expression patterns according to its subcellular location. GAGE7 exhibited nuclear, cytoplasmic and nucleolar expression patterns. Three cancer/testis antigens were also observed coordinately expressed in infiltrating ductal breast carcinoma. Patients with tumors with CT10 expression was significantly correlated with nodal metastases (P < 0.001) and advanced clinical stages (P = 0.001). Patients with tumors with cytoplasmic GAGE7 and with the expression of two or more cancer/testis antigens were significantly correlated with advanced clinical stages (P = 0.001 and P = 0.030). No signiï¬cant difference was identiï¬ed between the different expression patterns of CT45 and clinicopathologic variables. In addition, Kaplan-Meier analysis revealed that diffuse CT10 expression and coexpression of three cancer/testis antigens were related to the poor prognosis of patients with infiltrating ductal breast carcinoma. CONCLUSIONS: Diffuse CT10 expression and the coexpression of three cancer/testis antigens can be used as a biomarker to distinguish patients with a poorer outcome of the breast carcinoma. Our finding may provide useful data for evaluating the prognosis of this disease and improving the effectiveness of therapeutic application based on the three cancer/testis antigens.
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Antígenos de Neoplasias/metabolismo , Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/metabolismo , Carcinoma Ductal de Mama/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias de la Mama/patología , Carcinoma Ductal de Mama/patología , Femenino , Humanos , Metástasis Linfática , Persona de Mediana Edad , PronósticoRESUMEN
Hypertrophic scarring remains a major problem for patients who have suffered from surgeries or burns. Vascularization plays an important role in the early phase of hypertrophic scarring. Therefore, the inhibition of angiogenesis might be used as a preventive strategy. In this study, we assessed the effect of anti-angiogenesis resulting from adenovirus-mediated METH1 (metalloprotease and thrombospondin1) gene expression on the hypertrophic scar formation in a rabbit ear model of hypertrophic scarring. We first investigated the number of microvessel and microcirculatory perfusion in untreated scars on days 10, 30, 60, and 90 after epithelialization. Then, we examined the effect of anti-angiogenesis by adenovirus-mediated METH1 expression on hypertrophic scar formation by calculating the scar elevation index, counting the microvessel and argyrophilic nucleolar organizer region particle, and detecting the amount of collagen on days 30 and 60 after treatment. We found that untreated scar tissues at the proliferative phase (days 10-60 after epithelialization) had a significantly higher density of microvessel and microcirculatory perfusion than those at the mature phase (day 90 after epithelization) (both p<0.05). On days 30 and 60 after treatment, the hypertrophic scar formation was significantly inhibited in the treatment group. There was significantly reduced scar elevation index, microvessel count, number of argyrophilic nucleolar organizer region, and total collagen content for treated scars. Our results demonstrate that METH1 has a markedly inhibitive effect on the formation of hypertrophic scar, and may thus have a promising application in the prevention of human hyperthropic scars.
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Proteínas ADAM/fisiología , Cicatriz Hipertrófica/fisiopatología , Microcirculación , Neovascularización Patológica/fisiopatología , Piel/irrigación sanguínea , Proteínas ADAM/genética , Animales , Cicatriz Hipertrófica/prevención & control , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Neovascularización Patológica/prevención & control , Conejos , Úlcera Cutánea/genética , Úlcera Cutánea/fisiopatología , TransfecciónRESUMEN
OBJECTIVE: To investigate the effects of Notch signaling on scars in a rabbit ear model of hypertrophic scarring. METHODS: The hypertrophic scar of rabbits' ears was reproduced. The left rabbit's ear wounds as the N-[N-(3,5-difluorophenacetyl-L-alanyl)]-(S)-phenylglycine t-butyl ester (DAPT) treated group were treated intradermally with the gamma-secretase inhibitor DAPT to inhibit the activation of Notch at 1, 3, 7 and 14 day time points. The right ears as the control group were treated with normal saline at the same time points. Experimental and control wounds were harvested on days 14, 21, 28 and 35 post wounding, and then examined histologically to quantify hypertrophic index and fibroblasts. The expression of epidermal differentiation markers-keratin 14 (K14), keratin 19 (K19), Involucrin and Notch downstream molecules-P21, P63 were examined and analyzed with immunohistochemistry staining. RESULTS: Both hypertrophic index (1.93 +/- 0.32, 1.82 +/- 0.36, 1.79 +/- 0.25) and number of fibroblasts [(4.08 +/- 0.88), (3.30 +/- 0.53), (3.19 +/- 0.73) x 10(3)/mm(2)] in the DAPT treated group were significantly reduced on days 21, 28 and 35, compared with the control group [2.56 +/- 0.29, 2.61 +/- 0.30, 2.58 +/- 0.39, and (5.45 +/- 0.99), (4.80 +/- 1.13), (4.43 +/- 1.17) x 10(3)/mm(2), all P < 0.01)]. The K19, K14 and P63 increased their expression in the DAPT treated group (28.6% +/- 5.7%, 53.1% +/- 4.5%, 57.0% +/- 5.8%) relative to the control group (10.1% +/- 2.8%, 30.8% +/- 4.9%, 16.5% +/- 2.2%, all P < 0.01) on day 14 post wounding, while the Involucrin and P21 decreased their expression in the DAPT treated group (12.3% +/- 1.9%, 11.0% +/- 1.7%) relative to the control group (29.3% +/- 4.6%, 44.3% +/- 3.5%, both P < 0.01). CONCLUSION: Inactivation of Notch signaling will inhibit scar epidermis to over-differentiation, and thereby inhibit proliferation of hypertrophic scars in the rabbit ears.
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Cicatriz Hipertrófica/metabolismo , Cicatriz Hipertrófica/patología , Receptores Notch/metabolismo , Animales , Diferenciación Celular , Modelos Animales de Enfermedad , Femenino , Masculino , ConejosRESUMEN
OBJECTIVE: To study the expression of Notch receptors, ligands and downstream target genes in hypertrophic scar and normal skin, and to investigate its role in the development of hypertrophic scar. METHODS: By immunohistochemistry, the expression of epidermal differentiation markers- beta1 integrin, keratin 14 (K14) and keratin 19 (K19), as well as Notch 1-4 and Jagged1 were examined in hypertrophic scars and normal skins. The expression of Notch downstream genes- P21 and P63 was analyzed with real-time quantitative PCR and immunohistochemistry staining. RESULTS: Histological analysis revealed a significant epidermal thickening in the hypertrophic scars, with excessive cell layers above the basal layer. Compared to the normal epidermis, the expression of beta1 integrin, K19 and K14 decreased in hypertrophic scars (P <0.05). Positive expression rate of Notch1 and Jagged1 in keratinocytes was significantly higher in hypertrophic scar than in normal skin (P < 0.05), while there was no difference in Notch2 and 3 positive expression rate. Furthermore, the expression of P21 was significantly up-regulated, while the expression of P63 was down-regulated in keratinocytes of hypertrophic scar (P < 0.05). CONCLUSIONS: Notch signal may play an important role in hypertrophic scar pathogenesis. Over-differentiation of Keratinocytes in hypertrophic scar may be related to the overexpression of Notch1 and Jagged1, up-regulation of P21 gene and down-regulation of P63 gene.
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Cicatriz Hipertrófica/metabolismo , Epidermis/metabolismo , Receptor Notch1/metabolismo , Adulto , Proteínas de Unión al Calcio/metabolismo , Estudios de Casos y Controles , Cicatriz Hipertrófica/patología , Regulación hacia Abajo , Epidermis/patología , Femenino , Humanos , Integrina beta1/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Proteína Jagged-1 , Queratina-14/metabolismo , Queratina-19/metabolismo , Masculino , Proteínas de la Membrana/metabolismo , Proteínas Serrate-Jagged , Transducción de Señal , Regulación hacia Arriba , Adulto JovenAsunto(s)
Cariotipificación , Modelos Animales , Neoplasias/genética , Animales , Transformación Celular Neoplásica , Equidae , Padre , Femenino , Genoma , Caballos , Humanos , Masculino , Madres , Especificidad de la EspecieRESUMEN
OBJECTIVE: To investigate the effect of METH1 gene transfection on fibroblast proliferation and I, III collagen synthesis in rabbit ear scar. METHODS: The hypertrophic scar model on the rabbit ears was reproduced. 10 days after epithelization, Ad-METH1 was injected into the scar tissue. 30 days later, the effect of METH1 gene transfection on the angiogenesis, fibroblast proliferation and the ratio of collagen I/III in the scar tissue was detected by microcirculation microscope, AgNOR particle count and collagen dyeing. RESULTS: 30 days after injection of Ad-METH1, angiogenesis, fibroblast proliferation and the ratio of collagen I/III in the scar tissue were obviously suppressed. CONCLUSION: Early application of Ad-METH1 after epithelization can markedly inhibit the formation of the hypertrophic scar.
Asunto(s)
Proteínas ADAM/genética , Inhibidores de la Angiogénesis/genética , Cicatriz Hipertrófica/patología , Transfección , Animales , Cicatriz Hipertrófica/genética , Modelos Animales de Enfermedad , Oído/patología , Oído Externo/patología , Femenino , Masculino , Microcirculación , Neovascularización Patológica , Conejos , Cicatrización de HeridasRESUMEN
OBJECTIVE: To investigate the angiogenesis in hypertropic scar tissue of rabbit ears at different periods and to explore a new method to prevent hyperplastic scar. METHODS: Nineteen Japanese white rabbits (weigthing 2.0-2.5 kg) were made animal models of hypertropic scar of ear. At 10th, 30th, 60th and 90 days, after epithelization, the microvessel and microcirculation in hyperplastic scar of 8 rabbits were studied by microcirculation microscope and laser Doppler flowmetry. The other 11 rabbits' right or left ears were randomly chosen into experimental group and control group. At 10 days after epithelization, 40 microL of adenovirus extracellular protein with metalloprotease and thrombospondin 1 domains (Ad-METH1) was injected into tissue of scar along the perimeter of the scar in experimental group. The same volume of empty adenovirus was injected in control group. After 30 days of injection, the gross appearance of 10 rabbits' ears scar was recorded, the number of microvessel in scar was counted and HE stainning of scar tissue was performed in experimental and control groups. One additional rabbit was used to evaluate the mRNA and protein expression of METH1 by RT-PCR and Western blot after 3 days of injection. RESULTS: The average number of microvessel at 10, 30, 60 and 90 days after epithelization was 42.37 +/- 3.89, 49.46 +/- 4.13, 33.12 +/- 4.34 and 13.24 +/- 2.31, respectively; the average value of microcirculatory perfusion at 10, 30, 60 and 90 days after epithetlization was (37.75 +/- 2.11), (59.87 +/- 6.46), (44.53 +/- 6.14) and (29.21 +/- 1.84) PU; the density of microvessels and perfusion of microcirculation in scar tissues during proliferative stage (from 10 to 60 days after epithelization) were markedly higher than that during mature period (90 days after epithelization, P < 0.05). At 10 to 30 days after epithelization, the histoligical features of scar showed early stage of proliferation and proliferative stage appearance; at 60 days after epithelization, it is still in proliferative stage, while some of scars were in mature phase; at 90 days after epithelization, the histoligical features of scar were mature period appearance. At 3 days after Ad-METH1 injection, METH1 gene was successfully expressed at both mRNA and protein levels in experimental group, but not in control group. At 30 days after injection, the gross appearance observation showed that scars in experimental group were flat and soft with the color close to normal, but scars in control group were obvious and hard. The number of microvessel of scar tissue was 12.38 +/- 2.56 in experimental group and 48.12 +/- 6.46 in control group, showing statistically significant difference between two groups (P < 0.01). In experimental group, HE staining shows that the density of microvessel and the number of fibroblasts were greatly decreased and collagen fibers arranged regularly. In control group, plenty of fibroblasts and abundant microvessels were observed. Thick and tight collagen fibers were seen in the outer layer of dermis with a irregular arrangement. CONCLUSION: The anti-angiogenesis by Ad-METH1 may have a promising application in the prevention of human hyperthropic scar.
Asunto(s)
Proteínas ADAM/genética , Adenoviridae/genética , Inhibidores de la Angiogénesis/genética , Cicatriz Hipertrófica/patología , Neovascularización Patológica , Proteínas ADAM/metabolismo , Inhibidores de la Angiogénesis/metabolismo , Animales , Western Blotting , Cicatriz Hipertrófica/prevención & control , Modelos Animales de Enfermedad , Oído/patología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Conejos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , TransfecciónRESUMEN
OBJECTIVE: To investigate the effects of curcumin (Cur) and erythromycin (EM) on multidrug resistance (MDR) reversal of K562/A02 cell line and their mechanism. METHODS: MTT assay was employed to determine the sensitivity of Cur, EM-treated K562/A02 cells to adriamycin (ADM). Flow cytometry was used to measure intracellular mean fluorescence intensity (MFI) of daunorubicin (DNR). P-gp expression was determined by immunohistochemistry. RT-PCR technique was used to examine the mdr1 mRNA level. RESULTS: IC(50) of ADM in K562/A02 cells was decreased when treated with Cur or EM, and the reversal times (RvT) was 4.9, 3.7 respectively. The RvT reached to 11.3 when treated with Cur (2.5 microg/ml) combined with EM (120 microg/ml). The DNR MFI in K562/A02 cells was significantly lower than that in K562 cells (P < 0.01), and was increased significantly when treated with Cur (2.5 microg/ml) or EM (120 microg/ml) (P < 0.05). There was no significant difference between DNR MFI of K562/A02 cells treated with Cur (2.5 microg/ml) or EM (120 microg/ml). Immunohistochemistry showed that P-gp expression was significantly higher in K562/A02 cells than in K562 cells (P < 0.01), and was reduced in K562/A02 cells treated with each (P < 0.01), though being still higher than that in K562 cells (P < 0.01). P-gp expression of K562/A02 cells treated with each drug for 5 days were lower than that for 3 days (P < 0.01), and lowered further when treated with Cur and EM together (P < 0.01). Mdr1 mRNA level in K562/A02 cells was higher than in K562 cells (P < 0.01), and was decreased when treated with each of the drugs (P < 0.01). The mdr1 mRNA level of K562/A02 cells treated with Cur (2.5 microg/ml) plus EM (120 microg/ml) was decreased most significantly than that treated with other group of drugs. After 5 day treatment the mdr1 mRNA level of K562/A02 cells with Cur (2.5 microg/ml) was lower than that with EM 120 microg/ml (P < 0.01). CONCLUSIONS: Either Cur or EM can partly reverse the multidrug resistance of K562/A02 cells and decrease the expression and function of P-gp in a time-dependent way. MDR reversing effect of Cur combined with EM is stronger than that of Cur or EM alone.
Asunto(s)
Curcumina/farmacología , Resistencia a Múltiples Medicamentos/efectos de los fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Eritromicina/farmacología , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Antineoplásicos/farmacología , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Sinergismo Farmacológico , Epirrubicina/farmacología , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Inmunohistoquímica , Células K562 , Leucemia Eritroblástica Aguda/genética , Leucemia Eritroblástica Aguda/metabolismo , Leucemia Eritroblástica Aguda/patología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de TiempoRESUMEN
BACKGROUND: An unbalance of cell proliferation and cell apoptosis is an important mechanism in carcinogenesis, and angiogenesis also plays a crucial role in tumorigenesis. Recently, survivin has been identified as an important member of the inhibitor of apoptosis protein (IAP) family. Although it has been shown that survivin is highly expressed in gliomas, and is associated with tumorigenesis, progression, and poor prognosis of gliomas, as yet the relation of survivin expression with proliferation, apoptosis, and angiogenesis of gliomas it is still unclear. METHODS: Eighty-three cases of brain glioma were chosen and protein expressions of survivin and proliferating cell nuclear antigen (PCNA) in glioma cells and Factor VIII-related antigen (FVIII-RAg) in vascular endothelial cells were investigated by immunohistochemistry. Apoptotic cells of brain glioma were screened by TdT-mediated dUTP nick end-labeling (TUNEL), and survivin immunoreactivity score (IRS), proliferative index (PI), apoptotic index (AI), overall daily growth (ODG), and microvessel density (MVD) in brain gliomas were measured. RESULTS: The survivin IRS, PI, AI, ODG, and MVD of brain gliomas were 3.75 +/- 3.89, 28.39 +/- 19.49%, 1.00 +/- 0.80%, 12.19 +/- 10.21%, and 62.75 +/- 31.50, respectively, and all of them increased markedly with an increase in the pathologic grade of brain gliomas (P < 0.001 for all). PI, ODG, and MVD in the survivin-positive group were significantly higher than those in the survivin-negative group (P < 0.001 for all). PI, ODG, and MVD were positively correlated with survivin IRS (P < 0.001 for all). Although there was no significant difference between AI in the survivin-positive group or in the survivin-negative group (P = 0.108), AI was inversely correlated with survivin IRS (P = 0.005). CONCLUSIONS: Survivin is overexpressed in brain gliomas, which may play an important role in malignant proliferation, antiapoptosis, and angiogenesis of brain gliomas.