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1.
Biol. Res ; 53: 13, 2020. tab, graf
Article in English | LILACS | ID: biblio-1100919

ABSTRACT

BACKGROUND: Gallbladder cancer (GBC) is the most common tumor of the biliary tract. The incidence of GBC shows a large geographic variability, being particularly frequent in Native American populations. In Chile, GBC represents the second cause of cancer-related death among women. We describe here the establishment of three novel cell lines derived from the ascitic fluid of a Chilean GBC patient, who presented 46% European, 36% Mapuche, 12% Aymara and 6% African ancestry. RESULTS: After immunocytochemical staining of the primary cell culture, we isolated and comprehensively characterized three independent clones (PUC-GBC1, PUC-GBC2 and PUC-GBC3) by short tandem repeat DNA profiling and RNA sequencing as well as karyotype, doubling time, chemosensitivity, in vitro migration capability and in vivo tumorigenicity assay. Primary culture cells showed high expression of CK7, CK19, CA 19-9, MUC1 and MUC16, and negative expression of mesothelial markers. The three isolated clones displayed an epithelial phenotype and an abnormal structure and number of chromosomes. RNA sequencing confirmed the increased expression of cytokeratin and mucin genes, and also of TP53 and ERBB2 with some differences among the three cells lines, and revealed a novel exonic mutation in NF1. The PUC-GBC3 clone was the most aggressive according to histopathological features and the tumorigenic capacity in NSG mice. CONCLUSIONS: The first cell lines established from a Chilean GBC patient represent a new model for studying GBC in patients of Native American descent.


Subject(s)
Humans , Animals , Male , Middle Aged , Antigens, Tumor-Associated, Carbohydrate/genetics , Indians, South American/genetics , Gallbladder Neoplasms/genetics , Ascitic Fluid/metabolism , Tumor Cells, Cultured , Carcinogenicity Tests , Chile , DNA Fingerprinting , Tumor Suppressor Protein p53/genetics , Cisplatin/pharmacology , Mice, Inbred NOD , Clone Cells/drug effects , Clone Cells/metabolism , Sequence Analysis, RNA , Receptor, ErbB-2/genetics , Genes, erbB-2/genetics , Gene Expression Profiling , Cell Line, Tumor/drug effects , Cell Line, Tumor/metabolism , Deoxycytidine/analogs & derivatives , Deoxycytidine/pharmacology , Epithelial Cells/metabolism , Keratin-19/genetics , Keratin-7/genetics , Carcinogenesis/genetics , Gallbladder Neoplasms/metabolism , Antineoplastic Agents/pharmacology
2.
Biol. Res ; 52: 13, 2019. graf
Article in English | LILACS | ID: biblio-1011415

ABSTRACT

BACKGROUND: Ovarian cancer is a significant cancer-related cause of death in women worldwide. The most used chemotherapeutic regimen is based on carboplatin (CBDCA). However, CBDCA resistance is the main obstacle to a better prognosis. An in vitro drug-resistant cell model would help in the understanding of molecular mechanisms underlying this drug-resistance phenomenon. The aim of this study was to characterize cellular and molecular changes of induced CBDCA-resistant ovarian cancer cell line A2780. METHODS: The cell selection strategy used in this study was a dose-per-pulse method using a concentration of 100 µM for 2 h. Once 20 cycles of exposure to the drug were completed, the cell cultures showed a resistant phenotype. Then, the ovarian cancer cell line A2780 was grown with 100 µM of CBDCA (CBDCA-resistant cells) or without CBDCA (parental cells). After, a drug sensitivity assay, morphological analyses, cell death assays and a RNA-seq analysis were performed in CBDCA-resistant A2780 cells. RESULTS: Microscopy on both parental and CBDCA-resistant A2780 cells showed similar characteristics in morphology and F-actin distribution within cells. In cell-death assays, parental A2780 cells showed a significant increase in phosphatidylserine translocation and caspase-3/7 cleavage compared to CBDCA-resistant A2780 cells (P < 0.05 and P < 0.005, respectively). Cell viability in parental A2780 cells was significantly decreased compared to CBDCA-resistant A2780 cells (P < 0.0005). The RNA-seq analysis showed 156 differentially expressed genes (DEGs) associated mainly to molecular functions. CONCLUSION: CBDCA-resistant A2780 ovarian cancer cells is a reliable model of CBDCA resistance that shows several DEGs involved in molecular functions such as transmembrane activity, protein binding to cell surface receptor and catalytic activity. Also, we found that the Wnt/3-catenin and integrin signaling pathway are the main metabolic pathway dysregulated in CBDCA-resistant A2780 cells.


Subject(s)
Humans , Female , Ovarian Neoplasms/genetics , Gene Expression Regulation, Neoplastic/drug effects , Carboplatin/pharmacology , Drug Resistance, Neoplasm/genetics , Transcriptome/drug effects , Antineoplastic Agents/pharmacology , Ovarian Neoplasms/pathology , Ovarian Neoplasms/drug therapy , Phenotype , Signal Transduction , Cell Death/drug effects , Cell Death/genetics , Sequence Analysis, RNA , Cell Line, Tumor , Transcriptome/genetics
3.
Rev. méd. Chile ; 144(10): 1305-1318, oct. 2016.
Article in Spanish | LILACS | ID: biblio-845445

ABSTRACT

Pancreatic cancer is a malignancy of great impact in developed countries and is having an increasing impact in Latin America. Incidence and mortality rates are similar for this cancer. This is an important reason to offer to the patients the best treatments available. During the Latin American Symposium of Gastroenterology Oncology (SLAGO) held in Viña del Mar, Chile, in April 2015, a multidisciplinary group of specialists in the field met to discuss about this disease. The main conclusions of this meeting, where practitioners from most of Latin American countries participated, are listed in this consensus that seek to serve as a guide for better decision making for patients with pancreatic cancer in Latin America.


Subject(s)
Humans , Pancreatic Neoplasms/therapy , Adenocarcinoma/therapy , Practice Guidelines as Topic , Disease Management , Consensus Development Conferences as Topic , Deoxycytidine/analogs & derivatives , Deoxycytidine/therapeutic use , Chemoradiotherapy , Latin America , Antimetabolites, Antineoplastic/therapeutic use
4.
Biol. Res ; 49: 1-10, 2016. ilus, graf, tab
Article in English | LILACS | ID: lil-774432

ABSTRACT

BACKGROUND: Reprimo (RPRM), a highly glycosylated protein, is a new downstream effector of p53-induced cell cycle arrest at the G2/M checkpoint, and a putative tumor suppressor gene frequently silenced via methylation of its promoter region in several malignances. The aim of this study was to characterize the epigenetic inactivation and its biological function in BC cell lines. METHODS: The correlation between RPRM methylation and loss of mRNA expression was assessed in six breast cancer cell lines by methylation specific PCR (MSP), 5'-Aza-2'-deoxycytidine treatment and RT-PCR assays. MDA-MB-231 cells were chosen to investigate the phenotypic effect of RPRM in cell proliferation, cell cycle, cell death, cell migration and invasion. RESULTS: In the cancer methylome system (CMS) (web-based system for visualizing and analyzing genome-wide methylation data of human cancers), the CpG island region of RPRM (1.1 kb) was hypermethylated in breast cancer compared to normal breast tissue; more interesting still was that ERa(+) tumors showed higher methylation intensity than ERa(-). Downregulation of RPRM mRNA by methylation was confirmed in MDA-MB-231 and BT-20 cell lines. In addition, overexpression of RPRM in MDA-MB-231 cells resulted in decreased rates of cell migration, wound healing and invasion in vitro. However, RPRM overexpression did not alter cell viability, phosphatidylserine (PS) translocation or G2/M cell cycle transition. CONCLUSION: Taken together, these data suggest that RPRM is involved in decreased cell migration and invasion in vitro, acting as a potential tumor suppressor gene in the MDA-MB-231 cell line.


Subject(s)
Female , Humans , Breast Neoplasms/pathology , Cell Cycle Proteins/physiology , Cell Movement/physiology , Cell Proliferation/physiology , Glycoproteins/physiology , Analysis of Variance , Blotting, Western , Breast Neoplasms/genetics , Cell Cycle , Cell Line, Tumor , Cell Survival , Cell Cycle Proteins/genetics , Cell Movement/genetics , Cell Proliferation/genetics , DNA Methylation , Epigenesis, Genetic , Gene Expression Regulation, Neoplastic , Glycoproteins/genetics , Neoplasm Invasiveness , Real-Time Polymerase Chain Reaction , Statistics, Nonparametric
5.
Rev. méd. Chile ; 143(10): 1277-1285, oct. 2015. ilus, tab
Article in Spanish | LILACS | ID: lil-771711

ABSTRACT

Background: Endoscopic submucosal dissection (ESD) is a minimally invasive procedure that allows curative treatment of early gastric cancer (EGC) in selected patients. Aim: To report our initial experience with ESD. Material and Methods: Analysis of prospective data from 16 patients aged 61 to 84 years, who underwent ESD between December 2011 and June 2014. Tumor type, operative time, hospitalization length, oncologic outcomes, complications and short-term follow up were registered. Results: En-block resection was achieved in all cases. The median operative time was 135 min (range: 50-320 min). Specimens' median size was 3.5 cm (range: 3-10). All the resections were R0. In 14 patients ESD was considered curative. In two patients, ESD was considered potentially non-curative due to the presence pathological risk factors for lymph-node metastases in the biopsy specimen. Both patients underwent laparoscopic gastrectomy with lymph-node dissection. There was one case of gastric wall perforation that was repaired by laparoscopic suture. There was no mortality. The median follow-up time was 15 months (range: 2-30 months). Conclusions: ESD is a feasible and safe procedure in our institution with good results in this initial experience.


Subject(s)
Aged , Female , Humans , Male , Middle Aged , Adenocarcinoma/surgery , Dissection/methods , Gastrectomy/methods , Gastric Mucosa/surgery , Stomach Neoplasms/surgery , Adenocarcinoma/pathology , Disease-Free Survival , Early Detection of Cancer , Follow-Up Studies , Gastric Mucosa/pathology , Lymphatic Metastasis , Neoplasm Staging , Operative Time , Prospective Studies , Stomach Neoplasms/pathology , Treatment Outcome
6.
Int. j. morphol ; 29(3): 769-781, Sept. 2011. ilus
Article in Spanish | LILACS | ID: lil-608656

ABSTRACT

Las células madre mesenquimales son poblaciones multipotentes de células que tienen amplia capacidad de diferenciación, plasticidad y potencial inmunosupresor, lo que las hace una herramienta de gran importancia en las terapias basadas en células.En función de su potencial, se ha determinado que las células perivasculares poseen características de células madre de gran potencial clínico, no obstante, las propiedades biológicas que llevan a su diferenciación son menos comprendidas. Los últimos avances en la comprensión de la relación entre pericitos y las células madre mesenquimales plantean funciones específicas de tejido, así como, su potencial uso terapéutico en las isquemia, tales como, la cerebro-vascular y un mejor entendimiento de la vascularización patológica tumoral. A pesar de la creciente aceptación que las células perivasculares están relacionada o son células madre mesenquimales, existen escasas pruebas experimentales que muestren la diferenciación de pericitos en diferentes tipos de células (Feng et al., 2010). En esta revisión documentamos los fundamentos biológicos de los pericitos que respaldan su uso en terapia regenerativa.


Mesenchymal stem cells are a multipotent population of cells that have extensive capacity for differentiation, plasticity and immunosuppressive potential, making them an important tool in cell-based therapies. According to their potential, it has been determined that perivascular cells have stem cell characteristics of great clinical potential, however, the biological properties that lead to their differentiation are less understood. Recent advances in understanding the relationship between pericytes and mesenchymal stem cells pose tissue-specific functions, as well as their potential therapeutic use in ischemia, such as cerebro-vascular and a better understanding of the pathological tumor vascularization. Despite the growing acceptance that perivascular cells are related, or that they are mesenchymal stem cells, there is little experimental evidence to show the differentiation of pericytes in different cell types (Feng et al., 2010). In this review we document the biological basis of pericytes that support their use in regenerative therapy.


Subject(s)
Animals , Brain Neoplasms/therapy , Brain Neoplasms/ultrastructure , Pericytes/cytology , Pericytes/pathology , Pericytes/transplantation , Regenerative Medicine/methods
7.
Acta cir. bras ; 25(3): 225-230, May-June 2010. ilus, tab
Article in English | LILACS | ID: lil-546827

ABSTRACT

PURPOSE: To evaluate the results of a prospective therapeutic protocol with long-term follow up in terms of survival rates in a cohort of patients treated with Intermediate and Advanced GBC (GBC). METHODS: Prospective cohort of patients with intermediate and advanced stages of GBC treated between 1996 and 2006. All cases were treated with a partial hepatic segmentectomy on segments IVb and V and a regional lymph node dissection and six cycles of out-patient chemotherapy (5-FU and leukovorin). With an average follow-up of 31.5 months, the morbidity, operative mortality, hepatic and lymphatic infiltration and actuarial survival were measured. Descriptive statistics were applied as well as bivariate analysis applying Fisher's exact test and non-parametrical tests and Kaplan Meier survival curves. Also logistic regression and proportional risk of Cox were applied. RESULTS: 40 patients were included in this protocol, with an average age of 59.5 years (40-85 years), of which 28 were women (70 percent). Depth of wall infiltration: muscular 8 patients (20 percent), subserosal 12 patients (30 percent), serosal 12 patients (30 percent) and perivesicular adipose tissue 8 patients (20 percent). The series morbidity was 27.5 percent. There was no operative mortality. The chemotherapy was well tolerated. The overall actuarial survival in the series was 50 percent at 60 months. CONCLUSION: Our protocol treatment has morbidity, mortality and survival rates similar to previously reported series.


OBJETIVO: Avaliar os resultados de resultados da aplicação de um protocolo terapêutico de natureza prospectiva, com seguimento em longo prazo nos termos de taxas de sobrevivência em uma coorte de pacientes operados com carcinoma vesícula biliar (CVB) intermédio e avançado. MÉTODOS: A coorte prospectiva de pacientes com estágios intermediários e avançados de CVB tratados entre 1996 e 2006. Todos os casos foram tratados com uma segmentectomia hepática parcial em segmentos IVb e V e uma dissecção linfonodal regional e seis ciclos de quimioterapia de ambulatório (5-FU e leukovorin). Com um tempo de seguimento médio de 31,5 meses, a morbidade, mortalidade operatória, hepático e infiltração linfática e atuarial de sobrevida foram medidas. Estatísticas descritivas foram aplicadas, bem como análise bivariada aplicando o teste exato de Fisher, testes não-paramétricos, curvas de sobrevida Kaplan Meier e técnica de regressão logística e risco proporcional de Cox. RESULTADOS: Foram incluídos 40 pacientes neste protocolo, com uma média de idade de 59,5 anos (40-85 anos), dos quais 28 eram mulheres (70 por cento). Profundidade de infiltração parede: muscular 8 pacientes (20 por cento), subserosal 12 pacientes (30 por cento), serosas 12 pacientes (30 por cento) e perivesicular no tecido adiposo, 8 pacientes (20 por cento). A série morbidade foi de 27,5 por cento. Não houve mortalidade operatória. A quimioterapia foi bem tolerada. A sobrevida global atuarial da série foi de 50 por cento em 60 meses. CONCLUSÃO: Nosso protocolo tem tratamento morbidade, mortalidade e taxas de sobrevivência semelhantes às relatadas anteriormente série.


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Antineoplastic Agents/therapeutic use , Carcinoma/mortality , Carcinoma/therapy , Fluorouracil/therapeutic use , Gallbladder Neoplasms/mortality , Gallbladder Neoplasms/therapy , Clinical Protocols , Carcinoma/pathology , Chemotherapy, Adjuvant/methods , Chile/epidemiology , Cholecystectomy/methods , Epidemiologic Methods , Gallbladder Neoplasms/pathology , Leucovorin/therapeutic use , Neoplasm Invasiveness , Sex Factors , Treatment Outcome
8.
Rev. méd. Chile ; 136(11): 1381-1389, nov. 2008. graf, tab
Article in Spanish | LILACS | ID: lil-508957

ABSTRACT

Background: Human papillomavirus (HPV) infection is the most common sexually transmitted disease. Aim: To determine prevalence of HPV genital infection in voluntary asymptomatic male university students. Material and methods: A cross-sectional study in 62 asymptomatic, sexually active male students. Exfoliated cells were obtained from the penüe shaft and coronal sulcus. Samples were analyzed for HPV DNA detection and genotyping by polymerase chain reaction and Reverse Line Blot. Results: The prevalence of HPV infection was 84 percent. HPV detection was 77 percent in penile shaft and 66 percent in coronal sulcus. The most commonly detected types were HPV-16 (45 percent), HPV-11 (19 percent), HPV-6 (10 percent) and HPV-18 (9 percent). Múltiple infection wasfoundin 54 percent. The most frequent combinations were VPH11/16 (18 percent) and VPH16/18 (5 percent). Conclusions: HPV infection is highly frequent in asymptomatic male university students, high rísk HPV types were greatly predominant.


Subject(s)
Adult , Humans , Male , Middle Aged , Alphapapillomavirus , DNA, Viral/genetics , Papillomavirus Infections/epidemiology , Penis/virology , Students, Health Occupations/statistics & numerical data , Alphapapillomavirus/classification , Alphapapillomavirus/genetics , Alphapapillomavirus/isolation & purification , Cross-Sectional Studies , Chile/epidemiology , Genotype , Polymerase Chain Reaction , Prevalence , Papillomavirus Infections/virology , Sexual Behavior , Socioeconomic Factors
9.
Rev. méd. Chile ; 135(6): 768-772, jun. 2007. ilus
Article in Spanish | LILACS | ID: lil-459581

ABSTRACT

Hepatocarcinoma with sarcomatoid transformation is uncommon. It presents clinically with íeucocytosis and fever, resembling a liver abscess. We report a 40 year-old male that presented pain in the right upper quadrant and fever. Abdominal imaging showed an hypodense image in the liver that resembled a liver abscess. The patient was subjected to a percutaneous drainage obtaining 150 ml of an hemorrhagic fluid whose culture was negative. The clinical picture persisted and the patient was subjected to a right hepatectomy. The pathological study of the surgical piece disclosed a hepatocarcinoma with sarcomatoid transformation. The patient was discharged sixteen days after surgery.


Subject(s)
Adult , Humans , Male , Carcinoma, Hepatocellular/pathology , Liver Neoplasms/pathology , Biopsy , Cell Transformation, Neoplastic , Carcinoma, Hepatocellular/surgery , Drainage , Hepatectomy , Liver Abscess/diagnosis , Liver Neoplasms/surgery , Liver/pathology , Tomography, X-Ray Computed
10.
Rev. chil. cir ; 58(6): 441-446, dic. 2006. ilus, tab
Article in Spanish | LILACS | ID: lil-455711

ABSTRACT

Introducción: Se ha descrito variabilidad ambiental de Echinococcus granulosus (Eg). Se han reportado 10 genotipos y cierta heterogenicidad intergenotipo en estudios con material proveniente de animales. El objetivo de este estudio es describir los resultados de un protocolo de genotipificación de Eg en muestras de hidatidosis humana. Material y método: Estudio de corte transversal. Se recolectó el líquido hidatídico de una muestra consecutiva de pacientes intervenidos quirúrgicamente por hidatidosis hepática y pulmonar en hospitales de Temuco entre julio de 2004 y septiembre de 2005. Se diseñó un protocolo de extracción de ADN para Eg en muestras homogeneizadas de aspirado de quistes hidatídicos. Se emplearon 2 sistemas de reacción de polimerasa en cadena (PCR): PCREg9 y PCREg16; ambos en concentraciones de MgCl2 al 2mM. Para PCREg9, se utilizaron los primers Eg9F y Eg9R a concentraciones de 0.5 mM, empleándose 35 ciclos con temperatura de hibridación a 60°C. Para PCREg16 se utilizaron los primers Eg16F y Eg16R a concentraciones de 0,5 mM, empleándose 35 ciclos con temperatura de hibridación a 65°C. Los productos de las PCR fueron digeridos con una enzima de restricción (Rsa1) para la discriminación de los genotipos. Resultados: Se analizaron 25 muestras, 4 provenientes de quistes pulmonares y 21 de quistes hepáticos. Se logró amplificación de Eg en 22 de 25 muestras (88 por ciento). La digestión enzimática reveló la presencia de 3 genotipos posibles: en 21 de 22 muestras (95,45 por ciento) se observó un patrón de restricción correspondiente a los genotipos G1 ó G7 y en la muestra restante a los genotipos G4 ó G7. Conclusión: Con los sistemas de PCR empleados se detectó ADN de Eg.


Subject(s)
Male , Adult , Humans , Animals , Female , DNA , Echinococcus/genetics , Echinococcosis, Hepatic/genetics , Echinococcosis, Pulmonary/genetics , Polymerase Chain Reaction/methods , Cross-Sectional Studies , Genotype , Molecular Biology , Nucleic Acid Amplification Techniques , Species Specificity
11.
Int. j. morphol ; 21(3): 205-209, 2003. ilus
Article in Spanish | LILACS | ID: lil-388102

ABSTRACT

La anormalidad citogenética más común en la leucemia mieloide crónica (LMC) es el cromosoma Philadelphia, producida por la t(9;22), cuya expresión molecular es el gen de fusión BCR-ABL, que codifica proteínas con actividad tirosinquinasa. Según el punto de ruptura de los genes BCR o ABL se produce una proteína de fusión de 210-kD(p210) o 190-kD(p190). La presencia de este gen de fusión en pacientes con LMC tiene implicancia diagnóstica. Con el propósito de detectar transcriptos de fusión del gen BCR/ABL en pacientes con leucemia mieloide crónica, procedentes de la IX Región de Chile, se estudiaron 14 muestras de sangre de 11 pacientes con LMC. A 2 de ellos, se les realizó seguimiento durante su tratamiento con Gleevec. Se aplicó la técnica de reacción en cadena de la polimerasa con transcriptasa reversa (RT-PCR), usando una PCR en nido. Para la detección de los transcriptos de fusión p210 y p190 del gen BCR/ABL, se utilizaron 4 pares de iniciadores. En 9/14 muestras se detectó el transcripto de fusión p210 y en 5/14 los transcriptos de fusión p210 y p190. En los 2 pacientes en seguimiento, hubo desaparición del transcripto p190, permaneciendo el transcripto p210. Estos resultados reafirman la importancia de detectar transcriptos de fusión BCR/ABL para el diagnóstico y seguimiento durante el tratamiento de la LMC.


Subject(s)
Humans , Fusion Proteins, bcr-abl , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Transcription, Genetic/physiology , Philadelphia Chromosome
12.
Bogotá, D.C; s.n; mayo 1990. 73 p. tab, graf.
Thesis in Spanish | LILACS | ID: lil-189994

ABSTRACT

En este estudio de tipo transversal descriptivo, se realizaron 100 encuestas a diferentes médicos de Bogotá con el fin de conocer si utilizaban o no procedimientos médicos no ortodoxos (PMNO) y cuales eran estos procedimientos. En el análisis estadístico se estudiaron cuatro variables respecto al uso o no de estos procedimientos, que son: sexo, especialidad, edad y años de experiencia. Cruzando las diferentes variables se encontró que si hay relación entre el uso de PMNO y la edad y especialidad. No se encontró relación entre el uso de PMNO con el sexo ni con los años de experiencia del médico. Se realizó una revisión bibliográfica de algnunos procedimientos para conocer si realmente contenían principios activos que fueran demostrados científicamente. Se logró una reseña histórica en la que se evidenció, como desde el principio de la enseñanza médica en Colombia fue importante el conocimiento, uso y preparación de diferentes plantas medicinales


Subject(s)
Complementary Therapies
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