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1.
Biol Direct ; 18(1): 1, 2023 01 11.
Artículo en Inglés | MEDLINE | ID: mdl-36631876

RESUMEN

BACKGROUND: Pancreatic cancer (PC) is a highly lethal malignancy that requires effective novel therapies. M2 macrophages are abundant in the PC microenvironment and promote cancer progression. Exosomes are emerging mediators of the crosstalk between cancer cells and the microenvironment. This study was conducted to explore the role of M2 macrophage-derived exosomes in PC. METHODS: Exosomes derived from M2 macrophages were extracted. miR-193b-3p and TRIM62 were overexpressed or silenced to examine their function in PC. Luminescence assays were used to investigate the interaction between miR-193b-3p and TRIM62. Cell proliferation was examined by EdU staining. Would healing and transwell assays were applied to evaluate cell migration and invasion. Co-immunoprecipitation was used to assess the interaction between TRIM62 and c-Myc. Gene and protein expressions were analyzed by quantitative RT-PCR and immunoblotting, respectively. RESULTS: M2 macrophage-derived exosomal miR-193b-3p promoted the proliferation, migration, invasion, and glutamine uptake of SW1990 cells. Mechanism study revealed that TRIM62 is a target of miR-193b-3p. TRIM62 inhibited the proliferation, migration, invasion, and glutamine uptake of SW1990 cells by promoting c-Myc ubiquitination. Our data also suggested that TRIM62 expression negatively correlated with miR-193b-3p and c-Myc expression. High-expression of miR-193b-3p and c-Myc predicts poor prognosis, whereas low-expression of TRIM62 predicts poor prognosis in patients with PC. CONCLUSION: M2 macrophage-derived exosomal miR-193b-3p enhances the proliferation, migration, invasion, and glutamine uptake of PC cells by targeting TRIM62, resulting in the decrease of c-Myc ubiquitination. This study not only reveals the mechanism underlying the crosstalk between M2 macrophages and PC cells but also suggests a promising therapeutic target for PC.


Asunto(s)
Exosomas , MicroARNs , Neoplasias Pancreáticas , Humanos , MicroARNs/genética , MicroARNs/metabolismo , Glutamina/metabolismo , Neoplasias Pancreáticas/genética , Línea Celular Tumoral , Exosomas/genética , Exosomas/metabolismo , Exosomas/patología , Proliferación Celular , Macrófagos/metabolismo , Microambiente Tumoral , Neoplasias Pancreáticas
2.
BMC Genom Data ; 23(1): 5, 2022 01 10.
Artículo en Inglés | MEDLINE | ID: mdl-35012444

RESUMEN

BACKGROUND: Pancreatic cancer (PC) is a highly lethal disease and an increasing cause of cancer-associated mortality worldwide. Interferon regulatory factors (IRFs) play vital roles in immune response and tumor cellular biological processes. However, the specific functions of IRFs in PC and tumor immune response are far from systematically clarified. This study aimed to explorer the expression profile, prognostic significance, and biological function of IRFs in PC. RESULTS: We observed that the levels of IRF2, 6, 7, 8, and 9 were elevated in tumor compared to normal tissues in PC. IRF7 expression was significantly associated with patients' pathology stage in PC. PC patients with high IRF2, low IRF3, and high IRF6 levels had significantly poorer overall survival. High mRNA expression, amplification and, deep deletion were the three most common types of genetic alterations of IRFs in PC. Low expression of IRF2, 4, 5, and 8 was resistant to most of the drugs or small molecules from Genomics of Drug Sensitivity in Cancer. Moreover, IRFs were positively correlated with the abundance of tumor infiltrating immune cells in PC, including B cells, CD8+ T cells, CD4+ T cells, macrophages, Neutrophil, and Dendritic cells. Functional analysis indicated that IRFs were involved in T cell receptor signaling pathway, immune response, and Toll-like receptor signaling pathway. CONCLUSIONS: Our results indicated that certain IRFs could serve as potential therapeutic targets and prognostic biomarkers for PC patients. Further basic and clinical studies are needed to validate our findings and generalize the clinical application of IRFs in PC.


Asunto(s)
Factores Reguladores del Interferón , Neoplasias Pancreáticas , Humanos , Factores Reguladores del Interferón/genética , Neoplasias Pancreáticas/genética , Pronóstico , ARN Mensajero/genética , Neoplasias Pancreáticas
3.
BMC Cancer ; 20(1): 541, 2020 Jun 09.
Artículo en Inglés | MEDLINE | ID: mdl-32517802

RESUMEN

BACKGROUND: Recent evidence suggests that albumin-to-Alkaline Phosphatase Ratio (AAPR) functions as a novel prognostic marker in several malignancies. However, whether it can predict the prognosis of unresectable pancreatic ductal adenocarcinoma (PDAC) remains unclear. Herein, we seek to investigate this possibility by a propensity score matching (PSM) analysis. METHODS: This was a retrospective cohort study in which 419 patients diagnosed with unresectable PDAC and receiving chemotherapy were recruited. Patients were stratified based on the cutoff value of AAPR. The PSM analysis was performed to identify 156 well-balanced patients in each group for overall survival (OS) comparison and subgroup analysis. Univariate and multivariate analyses were carried out to examine the potential of AAPR to indicate the prognosis of unresectable PDAC. The prediction performance of conventional model and combined model including AAPR was compared using the Akaike Information Criterion (AIC) and concordance index (C-index). RESULTS: We identified an AAPR of 0.4 to be the optimal cutoff for OS prediction. Patients with AAPR≤0.4 had significantly shorter OS compared with patients with AAPR> 0.4 (6.4 versus 9.3 months; P < 0.001). Based on the PSM cohort and entire cohort, multivariate Cox analysis revealed that high pretreatment for AAPR was an independent marker predicting favorable survival in unresectable PDAC (hazard ratio, 0.556; 95% confidence interval, 0.408 to 0.757; P < 0.001). Significant differences in OS were observed in all subgroups except for the group of patients age ≤ 60. Combined prognostic model including AAPR had lower AIC and higher C-index than conventional prognostic model. CONCLUSIONS: Pretreatment AAPR servers as an independent prognostic indicator for patients with unresectable PDAC. Inclusion of AAPR improved the prediction performance of conventional prognostic model, potentially helping clinicians to identify patients at high risk and guide individualized treatment.


Asunto(s)
Fosfatasa Alcalina/sangre , Biomarcadores de Tumor/sangre , Carcinoma Ductal Pancreático/sangre , Neoplasias Pancreáticas/sangre , Albúmina Sérica/análisis , Adulto , Factores de Edad , Anciano , Anciano de 80 o más Años , Análisis de Varianza , Carcinoma Ductal Pancreático/tratamiento farmacológico , Carcinoma Ductal Pancreático/mortalidad , Distribución de Chi-Cuadrado , Intervalos de Confianza , Femenino , Humanos , Masculino , Persona de Mediana Edad , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/mortalidad , Pronóstico , Puntaje de Propensión , Estudios Retrospectivos , Factores Sexuales
4.
Aging (Albany NY) ; 12(1): 502-517, 2020 01 08.
Artículo en Inglés | MEDLINE | ID: mdl-31913856

RESUMEN

Chemokines play essential roles in the progression of various human cancers; however, the expression and role of CXC chemokines in pancreatic adenocarcinoma (PAAD) have not yet been identified. The aim of this study is to identify the expression patterns, clinical significance and mechanisms of CXC chemokines in regulating tumour microenvironment of PAAD. Three CXC chemokines, including CXCL5, CXCL9, and CXCL10, were significantly overexpressed in PAAD tissues, which were correlated with the poor survival of the patients. CXCL9/10 was associated with change of immune cell pattern in the tumour microenvironment, and supplementation of CXCL9 in the orthotopic murine PAAD model promoted tumour progression. In particular, CXCL9 reduced the CD8+ cytotoxic T lymphocytes in the tumour microenvironment of PAAD, which could be attributed to the reduced CD8+ T cell proliferation, activation, and secretion of anti-tumour cytokines. In vitro treatment of CXCL9 directly led to the suppression of the proliferation, activation, and secretion of anti-tumour cytokines of isolated CD8+ T cells. Inhibition of STAT3 recovered the CXCL9-inhibited proliferation, activation, and secretion of anti-tumour cytokines of CD8+ T cells. Our study indicates CXCL9 as a potential target of immunotherapy in PAAD treatment by regulating the CD8+ T lymphocytes in the tumour microenvironment.


Asunto(s)
Adenocarcinoma/inmunología , Adenocarcinoma/metabolismo , Quimiocina CXCL9/metabolismo , Neoplasias Pancreáticas/inmunología , Neoplasias Pancreáticas/metabolismo , Factor de Transcripción STAT3/inmunología , Linfocitos T Citotóxicos/inmunología , Linfocitos T Citotóxicos/metabolismo , Adenocarcinoma/mortalidad , Adenocarcinoma/patología , Animales , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Linfocitos T CD8-positivos/patología , Línea Celular Tumoral , Quimiocina CXCL9/genética , Quimiocina CXCL9/farmacología , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Expresión Génica , Humanos , Inmunomodulación , Inmunofenotipificación , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Linfocitos Infiltrantes de Tumor/metabolismo , Linfocitos Infiltrantes de Tumor/patología , Ratones , Neoplasias Pancreáticas/mortalidad , Neoplasias Pancreáticas/patología , Pronóstico , Proteínas Recombinantes/farmacología , Transducción de Señal , Linfocitos T Citotóxicos/efectos de los fármacos , Microambiente Tumoral/inmunología
5.
Cancer Manag Res ; 11: 3171-3185, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31114359

RESUMEN

Background: Accumulating studies have reported that aberrant expression of SLC5A1 is a negative prognostic factor to various cancer patients. Purpose: Pancreatic cancer tissue has also shown to harbor higher expression of SLC5A1, however how SLC5A1 mediates pancreatic cancer cells growth remains unclear. Methods: In this study, we examined the mRNA and protein expressions of SLC5A1 in human pancreatic tissue and various cell lines. The in vitro and in vivo roles of SLC5A1 in pancreatic cancer were investigated through stably transfected pancreatic cells with shRNA plasmid targeting SLC5A1. Results: Our results observed SLC5A1 was over-expressed in human pancreatic cancer tissues as well as most pancreatic cancer cell lines. Both in vitro and in vivo inhibition of SLC5A1 retarded pancreatic cancer cell growth and progression. The SLC5A1 knockdown mediated growth suppression is mainly regulated by reduced cellular glucose uptake by pancreatic cancer cells. Our further mechanistic observation showed that inhibition of SLC5A1 induced AMPK-dependent mTOR suppression and pharmacological inhibition of AMPK rescued the effect of SLC5A1 blockade. Further protein-protein interaction analysis showed association of SLC5A1 with EGFR and knockdown of EGFR also showed decreased cellular survival and glucose uptake by pancreatic cancer cells. Conclusion: Our findings postulated SLC5A1/EGFR as the potential therapeutic target of pancreatic cancer patients.

6.
Biochem Biophys Res Commun ; 511(2): 482-489, 2019 04 02.
Artículo en Inglés | MEDLINE | ID: mdl-30808544

RESUMEN

Pancreatic cancer is a highly malignant tumor of the digestive system. Previous studies have shown that abnormal cell surface glycosylation is associated with cancer metastasis, which suggests that glycosylation changes may open a new window for discovering metastasis-related pathways. In this study, we used a microarray with 55 lectins to screen for altered glycosylation between two metastatic pancreatic cancer lines (Capan-1 and Su.86.86) and two nonmetastatic pancreatic cancer lines (Panc-1 and MIA PaCa-2), and we further analyzed three lectins with high-binding activities (AAL, UEA-I, and PHA-E) in cell motility assays using these pancreatic cancer cells to detect whether blocking certain forms of cell surface glycosylation affects any processes associated with metastasis. As a result, we found that AAL, a fucose-specific lectin, has different binding patterns between metastatic pancreatic cancer and nonmetastatic pancreatic cancer lines and inhibits cell motility in metastatic pancreatic cancer cells. Furthermore, the N-fucosylation-related genes FUT3, 5, and 6 were found to be responsible for the elevated fucosylation in metastatic pancreatic cells through real-time PCR screening. In summary, our findings that the specific bindings of AAL on cell surfaces and highly expressed FUT3, 5, and 6 in metastatic pancreatic cancer cells, although preliminary, are encouraging, and our established combined method is also suitable for discovering metastasis-related mechanisms in other cancers.


Asunto(s)
Fucosiltransferasas/genética , Neoplasias Pancreáticas/genética , Línea Celular Tumoral , Movimiento Celular , Fucosa/genética , Regulación Neoplásica de la Expresión Génica , Humanos , Invasividad Neoplásica/genética , Metástasis de la Neoplasia/genética , Regulación hacia Arriba
7.
Pancreatology ; 19(2): 346-351, 2019 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-30638854

RESUMEN

OBJECTIVE: The aim of this study was to evaluate the prognostic value of pre-treatment plasma hemostatic parameters in patients with advanced pancreatic cancer. METHODS: A total of 320 patients diagnosed with advanced pancreatic cancer between January 1, 2011 to December 31, 2015 were enrolled in this retrospective study. The prognostic significance of hemostatic parameters such as prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FBG), platelet count (PLT), mean platelet volume (MPV), plateletcrit (PCT), and platelet distribution width (PDW) was determined by univariate and multivariate Cox hazard models. Then, Kaplan-Meier methods and log-rank tests were performed to compare the survival of patients in different risk groups. RESULT: Univariate and multivariate analyses showed that prolonged PT, high FBG, and high MPV were independent prognostic factors for poor overall survival (PT > 11.3 s, HR = 1.46, 95%CI = 1.10-1.94, p = 0.009; FBG>2.5 g/L, HR = 1.41, 95%CI = 1.08-1.84, p = 0.011; MPV>12.2 fL, HR = 1.52, 95%CI = 1.13-2.04, p = 0.005). Moreover, all the patients were stratified into three groups by a scoring system based on these three hemostatic markers. The median survival time of the three groups was 8.8 months, 6.3 months and 4.3 months (P < 0.001). CONCLUSION: PT, FBG and MPV were independent prognostic factors in advanced pancreatic cancer. A novel scoring system based on these hemostatic parameters could be used to predict the survival of patients with advanced pancreatic cancer.


Asunto(s)
Fibrinógeno , Neoplasias Pancreáticas/clasificación , Neoplasias Pancreáticas/patología , Tiempo de Tromboplastina Parcial , Recuento de Plaquetas , Tiempo de Protrombina , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor , Estudios de Cohortes , Femenino , Humanos , Masculino , Volúmen Plaquetario Medio , Persona de Mediana Edad , Análisis Multivariante , Pronóstico , Estudios Retrospectivos , Factores de Riesgo
8.
J Ethnopharmacol ; 183: 143-150, 2016 May 13.
Artículo en Inglés | MEDLINE | ID: mdl-26934449

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Intestinal mucositis induced by chemotherapy is a severe clinical problem in cancer patients that currently lack effective interventions. In traditional Chinese medicine, chemotherapeutic toxicity is diagnosed as Qi and Yin deficiency, and steamed rehmannia root (SRR) is frequently prescribed to these patients. Whether SRR can prevent the adverse effects remains to be confirmed experimentally. The present study used a rat model to investigate potential efficacy and action mechanisms of SRR in attenuating the adverse effects caused by chemotherapy. MATERIALS AND METHODS: Intraperitoneal injection of a single dose of anti-metabolite methotrexate (MTX, 25mg/kg) was given to adult Wistar rats, which also received oral gavage of water or SRR (1.08g/kg twice daily 3 days before and 4 days after MTX treatment), or calcium folinate (CF, a clinically used MTX antidote as a comparison, at 1mg/kg twice daily 36h after MTX treatment), or SRR and CF in combination. Animals were sacrificed 4 days after MTX treatment. Complete blood cell counting was carried out. Jejunum was analyzed histologically for mucosal damage, immunohistochemically for proliferating cell nuclear antigen (PCNA), and biochemically for thiobarbituric acid reactive substances (TBARS) and reduced glutathione (GSH), as well as for tumor necrosis factor alpha (TNF-α). RESULTS: MTX treatment led to weight loss, leucopenia, polycythemia, increase in large thrombocyte ratio, intestinal villus atrophy, crypt loss and reduction in PCNA positive crypt cells, increases in mucosal TBARS and TNF-α and decrease in GSH. All these alterations were inhibited by SRR administration except leucopenia, and the effects of CF or CF plus SRR supplementation were found to be inferior to those of SRR. CONCLUSIONS: SRR can alleviate MTX-induced gut mucositis, which may be achieved by inhibiting MTX-induced oxidative stress and inflammatory response. These findings support the application of SRR in chemotherapy but not the combined application of SRR and CF.


Asunto(s)
Mucosa Intestinal/efectos de los fármacos , Metotrexato/efectos adversos , Mucositis/inducido químicamente , Mucositis/tratamiento farmacológico , Raíces de Plantas/química , Plantaginaceae/química , Rehmannia/química , Animales , Modelos Animales de Enfermedad , Femenino , Glutatión/metabolismo , Mucosa Intestinal/metabolismo , Mucositis/metabolismo , Preparaciones de Plantas/química , Preparaciones de Plantas/farmacología , Ratas , Ratas Wistar , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Pérdida de Peso/efectos de los fármacos
9.
PLoS One ; 9(8): e101536, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25170868

RESUMEN

BACKGROUND: Hepatocellular carcinoma (HCC) can be diagnosed by noninvasive approaches with serum α-fetoprotein (AFP) levels >200 ng/ml and/or a radiological imaging study of tumor mass >2 cm in patients with chronic liver disease. Percutaneous fine needle aspiration (FNA) under ultrasound (US) guidance has a diagnostic specificity of 95% and is superior to radiological imaging studies. AIM: The aim of this study is to elucidate the effectiveness and complications of fine needle aspiration in a Chinese population with primary liver cancer and AFP levels ≤200 ng/ml. MATERIALS AND METHODS: A retrospective study was conducted over a period of 28 years. This selection period included patients with a suspected diagnosis of primary liver cancer whose AFP levels were ≤200 ng/ml and who underwent US-FNA. This data was then analyzed with cytomorphological features correlating with medical history, radiological imaging, AFP, and follow-up information. RESULTS: Of the 1,929 cases with AFP ≤200 mg/ml, 1,756 underwent FNA. Of these, 1,590 cases were determined malignant and the remaining 166 were determined benign. Further, 1,478 malignant cases were diagnosed by FNA alone, and of these, 1,138 were diagnosed as PLC. The sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy of the diagnoses were 92.96%, 100%, 100%, 59.71%, and 93.62% respectively. There was no significant difference in the sensitivity, specificity, PPV and NPV between the subgroups with tumor size<2 cm and ≥2 cm. Major complications included implantation metastasis and hemorrhage. CONCLUSION: Patients with PLC, especially those who present with an AFP ≤200 ng/ml, should undergo FNA. If negative results are obtained by FNA, it still could be HCC and repeated FNA procedure may be needed if highly suspicious of HCC on imaging study. The superiority of FNA in overall accuracy may outweigh its potential complications, such like hemorrhage and implantation metastasis.


Asunto(s)
Carcinoma Hepatocelular/patología , Biopsia por Aspiración con Aguja Fina Guiada por Ultrasonido Endoscópico , Neoplasias Hepáticas/patología , Hígado/patología , alfa-Fetoproteínas/análisis , Carcinoma Hepatocelular/sangre , Biopsia por Aspiración con Aguja Fina Guiada por Ultrasonido Endoscópico/efectos adversos , Biopsia por Aspiración con Aguja Fina Guiada por Ultrasonido Endoscópico/métodos , Femenino , Humanos , Neoplasias Hepáticas/sangre , Masculino , Estudios Retrospectivos
10.
BMC Gastroenterol ; 14: 70, 2014 Apr 09.
Artículo en Inglés | MEDLINE | ID: mdl-24712400

RESUMEN

BACKGROUND: Current literature has demonstrated that host glutamine depletion facilitates tumorigenesis. Likewise, the glutamine transporter SLC38A1 is putatively associated with malignant transformation and tumor progression. Taken together, this forms the premise for undertaking the current study. The twofold aim of this study was to provide insight into whether or not a variance in the expression of SLC38A1 exists between human gastric cancer and healthy human tissues, and to determine how silencing the SLC38A1 gene could affect the proliferation, viability, migration, and invasion of gastric cancer cells. METHODS: Immunohistochemical staining was used to analyze the expression of SLC38A1 in gastric cancer tissues and adjacent healthy mucosa in 896 patients with pathologically confirmed gastric cancer who had underwent R0 resection. SH-10-TC cells (a gastric cancer cell line) were used to examine whether silencing SLC38A1 with siRNA could affect cell viability, migration and invasion. RESULTS: The SLC38A1 protein was very low or undetectable in healthy gastric mucosa. In contrast, strong staining of SLC38A1 protein was found in the cytoplasm in 495 out of the 896 gastric cancer samples. More pronounced SLC38A1 expression in gastric cancer tissues was significantly associated with age, differentiation status, lymph node metastasis, TNM stage and PCNA (proliferating cell nuclear antigen) expression. Upon univariate survival analysis, SLC38A1 expression was correlated with poor survival. Multivariate survival analysis revealed that SLC38A1 was an independent prognostic factor. CONCLUSION: SLC38A1 is overexpressed in gastric cancer, which suggests that it is contributory to tumor progression. These results encourage the exploration of SLC38A1 as a target for intervention in gastric cancer.


Asunto(s)
Adenocarcinoma/metabolismo , Sistema de Transporte de Aminoácidos A/metabolismo , Pueblo Asiatico , Neoplasias Gástricas/metabolismo , Adenocarcinoma/mortalidad , Sistema de Transporte de Aminoácidos A/genética , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Pronóstico , ARN Interferente Pequeño , Neoplasias Gástricas/mortalidad
11.
Hepatogastroenterology ; 60(128): 1906-10, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-24088318

RESUMEN

BACKGROUND/AIMS: To evaluate the safety and efficacy of high intensity focused ultrasound (HIFU) therapy in patients with local advanced pancreatic cancer. METHODOLOGY: 39 patients with local advanced pancreatic cancer were treated with HIFU, including 26 male and 13 female patients. The locations of the tumours were as follows: head of pancreas in 7 patients, body and/or tail of pancreas in 32 patients. Pain relief, time to progression (TTP), median survival and complications were analysed after HIFU treatment. RESULTS: There were no severe complications or adverse events related to HIFU therapy in any of the patients treated. Pain relief was achieved in 79.5% of patients. Median TTP was 5.0 months. The median overall survival time was 11 months. 6-month and 1-year survival rate for patients were 82.1% and 30.8% respectively. CONCLUSIONS: Although this study may have limitations, preliminary results demonstrate the safetyof clinical application of HIFU for pancreatic cancer and reveal it to be a promising mode of treatment for local advanced pancreatic cancers.


Asunto(s)
Ultrasonido Enfocado de Alta Intensidad de Ablación , Neoplasias Pancreáticas/cirugía , Adulto , Anciano , Progresión de la Enfermedad , Femenino , Ultrasonido Enfocado de Alta Intensidad de Ablación/efectos adversos , Ultrasonido Enfocado de Alta Intensidad de Ablación/mortalidad , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Dolor/etiología , Dolor/prevención & control , Neoplasias Pancreáticas/complicaciones , Neoplasias Pancreáticas/mortalidad , Neoplasias Pancreáticas/patología , Estudios Prospectivos , Análisis de Supervivencia , Factores de Tiempo , Tomografía Computarizada por Rayos X , Resultado del Tratamiento
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