RESUMEN
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.
Asunto(s)
Humanos , Femenino , Neoplasias Ováricas/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Carboplatino/farmacología , Resistencia a Antineoplásicos/genética , Transcriptoma/efectos de los fármacos , Antineoplásicos/farmacología , Neoplasias Ováricas/patología , Neoplasias Ováricas/tratamiento farmacológico , Fenotipo , Transducción de Señal , Muerte Celular/efectos de los fármacos , Muerte Celular/genética , Análisis de Secuencia de ARN , Línea Celular Tumoral , Transcriptoma/genéticaRESUMEN
Background: Chlamydia trachomatis infection is the most commonly reported sexually transmitted bacterial infection worldwide. Between 70 and 90% of women are asymptomatic, however, untreated and persistent infections can lead to the development of urethritis, pelvic inflammatory disease, infertility and ectopic pregnancy. Aims: To determine C. trachomatis infection frequency in a group of women in Chile, using quantitative real time PCR (qPCR) and to compare the usefulness of endocervical and urine samples for C. trachomatis detection. Methods: 87 asymptomatic women aged 15-64 years were included. Every woman donated one endocervical sample and one urine sample. Detection and quantification of C. trachomatis was performed by qPCR. Results: Of 87 endocervical samples, the frequency was 11.49% (n = 10). Of these samples, 5 cases were found in women < 35 years old. About urine samples, 16 samples were positive (18.39%). Ten women < 35 years old yielded positive urine samples. Only four women had both samples positive for C. trachomatis (4.6%). There was no statistically significant relationship between age and C. trachomatis infection. Cryptic plasmid quantification was found between 3.55 - 96.050 copies/μL for endocervical samples and 7.22-633.1 copies/μL for urine samples. Conclusion: Estimated frequency of C. trachomatis in Chilean women was higher than previous Chilean studies. Both types of samples are complementary for screening and diagnosis strategies using sensitive techniques, because silent infection can be present in either urinary or genital tract or in both in women.
Introducción: La infección por Chlamydia trachomatis es la infección bacteriana de transmisión sexual más frecuente en el mundo. Entre 70 y 90% de las mujeres son asintomáticas; sin embargo, las infecciones sin tratar y persistentes permiten el desarrollo de uretritis, enfermedad inflamatoria pélvica, infertilidad y embarazo ectópico. Objetivos: Determinar la frecuencia de infección por C. trachomatis en un grupo de mujeres chilenas, mediante RPC en tiempo real cuantitativa (qPCR) y comparar la utilidad de muestras endo-cervicales y de orina para la detección de C. trachomatis. Metodología: Participaron 87 mujeres asintomáticas (15-64 años). Cada mujer donó una muestra endo-cervical y una de orina. Se realizó detección y cuantificación C. trachomatis mediante qPCR. Resultados: La frecuencia de infección por C. trachomatis en muestras endo-cervicales fue de 11,49% (n: 10) y en muestras de orina de 18,39% (n: 16). El mayor número de casos se encontró en mujeres < 35 años. Sólo en cuatro mujeres se detectó C. trachomatis en ambas muestras (4,6%). La cuantificación de plásmido críptico se encontró en un rango 3,55 - 96.050 copias/μL. Conclusión: La frecuencia estimada de C. trachomatis fue más alta que en otros estudios chilenos. Ambos tipos de muestra deberían ser complementarias para estrategias de tamizaje y diagnóstico de C. trachomatis usando técnicas sensibles de detección, ya que la infección puede desarrollarse en el tracto genital y/o en el tracto urinario en mujeres.