Functional and genomic characterization of three novel cell lines derived from a metastatic gallbladder cancer tumor

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.

Score genético predice agresividad del cáncer de próstata / Genetic score predicts aggressiveness of prostate cancer

Establecer un score genético utilizando los polimorfismos de nucleótido único (SNPs) del gen que codifica para Ribonucleasa L (RNASEL)y regiones cromosómicas 8q24 y 17q12-24 en combinación con el antígeno específico de la próstata (PSA) para predecir la agresividad del cáncer de próstata (CaP). Pacientes y métodos: hombres con CaP tratados con prostatectomía radical. Se analizaron variables clínicas y patológicas: edad al diagnóstico, PSA al diagnóstico, el volumen tumoral (TV) y extensión extracapsular (ECE) según el TNM (tumour, node and metastasis) (ECE ≥T3) y score de Gleason. Desarrollamos un modelo de puntaje genético usando regresión logística multivariable. Resultados: se incluyeron 86 pacientes sometidos a prostatectomía radical. Edad promedio fue de 62 ± 7,5 años. El promedio de PSA fue de 11,3 ± 10,6 ng/mL. Treinta y un pacientes (36 por ciento) tuvieron ECE. La mediana del TV fue de 3,8 cc. Un PSA ≥ 10 ng/mL se asoció con una mayor tasa de ECE (p <0,05) y TV más alto (p = 0,032). En el análisis univariable, los pacientes con > 1 SNP tienen mayor riesgo de ECE que los pacientes con ≤ 1 SNP (42 por ciento vs. 10,5 por ciento, p = 0,01), y los pacientes con ≥ 3 SNP tienen más TV que los pacientes con <3 SNP (60 por ciento vs. 32 por ciento, p = 0,015). Se crearon dos modelos de riesgo usando el número de SNP y PSA ≥ o <10 ng/mL para predecir ECE (sensibilidad 67 por ciento y especificidad 84 por ciento) y TV (sensibilidad 59 por ciento y especificidad 70 por ciento). Conclusiones: El score genético presentado en este estudio es una herramienta novedosa para predecir indicadores de agresividad del CaP, como ECE y TV.(AU)

To establish a genetic score using SNPs (from RNAsel and chromosomal regions 8q24 and 17q12-24) in combination with Prostate Specific Antigen (PSA) at diagnosis to predict aggressiveness of PCa (tumor volume (TV) and extracapsular extension (ECE)). Patients and methods: Men with PCa diagnosed by needle biopsy and treated with radical prostatectomy (RP). Clinical and pathological variables such as age at diagnosis, PSA at diagnosis, TV, extension of tumor according TNM (ECE ≥T3) and Gleason score where analyzed. We developed a genetic score model using Multivariate Logistic Regression. Results: We included 86 patients who underwent RP. Mean age 62 ± 7.5 years. Mean PSA was 11.3 ± 10.6 ng/mL. Thirty-one patients (36 percent) had ECE. Median TV was 3.8 cc. PSA ≥ 10 ng/mL was associated with increased rate of ECE (p <0.05) and higher TV (p = 0.032). In univariate analysis, patients with more than 1 SNP had a greater risk of ECE than patients with ≤ 1 SNP (42 percent vs. 10.5 percent, p = 0.01), and patients with ≥ 3 risk SNPs had more TV than patients with <3 SNPs risk (60 percent vs. 32 percent, p = 0.015). Two models of risk using the number of SNPs and PSA ≥ or <10 ng/mL to predict ECE (sensitivity 67 percent and specificity 84 percent) and TV (sensitivity 59 percent and specificity 70 percent) were created. Conclusions: Genetic score usingdescribed SNPs and preoperative PSA can predict aggressiveness of PCa, which would be useful to define a management with more information at diagnosis especially in localized cancers.(AU)

Estilo de vida y agresividad del cáncer de próstata / Lifestyle and aggressiveness of prostate cancer

El objetivo de este estudio fue establecer una asociación entre diversas variables demográficas y epidemiológicas con la agresividad del cáncer de próstata (CaP). Métodos: pacientes diagnosticados con CaP respondieron una encuesta que incluye el nivel de educación, los factores de riesgo cardiovascular (FRCV), los antecedentes familiares (HF) de CaP, consumo de alcohol, tabaquismo y otros. Se utilizó análisis univariado y multivariado (AMV) para establecer si los factores mencionados anteriormente afectan las variables asociadas con la agresividad del CaP, como la edad al momento del diagnóstico, el índice de Gleason, los márgenes positivos (MP) y las metástasis óseas (MO), entre otras. Resultados: se incluyeron ciento setenta y dos hombres en el análisis. Los pacientes con HF fueron diagnosticados a edades más tempranas que los pacientes sin HF (55,73 vs 66,45 años, p = 0,0001). Los pacientes que beben tienen un mayor número de MP que los pacientes que no (15 vs 4 pacientes, p = 0,04). El AMV mostró que los pacientes que consumen alcohol y los que fuman (activos o suspendidos) tuvieron un mayor riesgo de MP (OR = 4,45 y 4,1, IC 95 por ciento 1,16-17,07 y 1,14-14,72, respectivamente, ambos p <0,05). Los pacientes con mayor nivel de educación presentaron un mayor riesgo de CaP confinado (OR = 3,42, IC 95 por ciento 1,392-8,434, p = 0,007). Conclusiones: los pacientes que consumen alcohol, fuman y tienen un menor nivel de educación presentaron un mayor riesgo de desarrollar CaP agresivo. (AU)

The aim of this study was to establish an association between various demographic and epidemiological variables with aggressiveness of prostate cancer (PCa). Methods: Patients diagnosed with PCa, answered a survey that include level of education, cardiovascular risk factors (CVRF), family history (FH) of PCa, alcohol intake, smoke and others. Univariate and multivariate analysis (MVA) were used to establish whether the factors mentioned above affect variables associated with aggressiveness of PCa such as: age at diagnosis, Gleason score, positive margins (PM), and bone metastasis (BM). Results: One hundred and seventy two men were included in the analysis. Patients with FH had cancer diagnosed at younger ages (55.73 years to FH vs 66.45 years to no FH, p = 0.0001). Patients who drink had higher number of PM than patients who did not (15 vs 4 patients, p= 0.04). MVA showed that patients who consumed alcohol and patients who smoked (active or suspended) had an increased risk of PM (OR= 4.45 and 4.1, 95 percent CI 1.16-17.07 and 1.14-14.72, respectively, both p<0.05). Patients with higher level of education presented an increased risk of confined PCa (OR= 3.42, 95 percent CI 1.392-8.434, p= 0.007). Conclusions: Patients who consume alcohol, smoke and have lower level of education presented a higher risk of developing aggressive PCa. (AU)

Obtención de cultivos primarios de estroma prostático benigno y tumoral mediante explantes de tejido desde biopsias por punción / Obtaining primary cultures of bening and tumoral prostatic stromal tissue through explants from needle biopsias

La progresión del cáncer de próstata (CaP) es regulada por el microentorno tumoral,el cual tiene como principal componente el estroma asociado al carcinoma (CAS). Sin embargo, los métodos experimentales usando cultivos primarios para modelar el comportamiento del estroma tumoral han sido insatisfactorios debido a la dificultad de obtener cultivos primarios de células estromales prostáticas humanas (Hpscs) con alta proporción de CAS. Así, el objetivo de este estudio consistió en estandarizar una metodología que permita ta obtención de cultivos primarios de HPSCs con alta proporción de CAS. Métodos: se recolectaron biopsias prostáticas de 10 pacientes,5 de ellos con CaP localmente avanzado y/o metastásico, y 5 sin evidencia de neoplasia. Se evaluó la proporción estromal por estudios histológicos y se estandarizó la obtención de cultivos primarios de HPScs mediante explantes de tejido. Los culilvos se caracterizaron por curvas de crecimiento y proliferación Resultados: El tejido obtenido desde biopsias prostáticas por punción de pacientes con CaP localmente avanzado y/o metastásico presentan alta proporción de CAS. La técnica explantes de tejido permite la obtención de cultivos primarios de HPSC desde biopsias, indicando además que cultivos primarios de CAS presentan un patrón de crecimiento y proliferación, superior a las células obtenidas de tejido benigno (BAS). Discusión: Nuestro estudio demuestra que a través de la técnica explantes de tejido es posible obtener cultivos primarios de HPSCs con CaP invasor, debido a que estos presentan zonas con alta proporción de CAS...

Tumor microenvironment plays a critical role in the progression of prostate cancer (CaP), which main component ¡s the ®carcinoma associated stroma¼ (CAS).However, the in vitro models utilizing human prostate stroma cells (HPSCs) as primary cultures have failed in recapitulate the human prostate microenvironment due to the unfeasibility to obtain primary cultures of HPSCs with a pure population of CAS. The aim of this study was to standardize a new methodology that allow to obtain primary cultures of HPSCs with high proportion of CAS. Methods: Primary cultures of HPSCs were obtained from explants of human needle biopsies from 5 benign and 5 locally advances and/or metastatic human prostate tissues. The primary cell cultures were characterized by proliferation rates and growth curves. Locally advanced and/or metastatic prostate needle biopsies exhibit a high presence of CAS. The analysis of human primary cultures isolated from benign and malignant prostate tissue explants revealed distinctive populations of HPSCs that can be distinguishable by proliferation rates and growth curves. Discussion: our studies demonstrated for the first time that prostate explants from needle biopsy, represents a feasible technique to obtain primary cultures of stromal cells from benign and malignant tissues, and represents in more accurate way the complexity of the tumor microenvironment...