ABSTRACT
Introducción: El virus del papiloma humano de alto riesgo (VPH-AR) es responsable del cáncer de cuello uterino y sus lesiones preneoplásicas. Los genotipos VPH16 y VPH18 son los más frecuentes en este cáncer. La integración del VPH-AR en el genoma de la célula hospedera es crucial en la carcinogénesis cervical, pero la etapa en que ocurre en la población chilena es incierta. Objetivo: Evaluar la integración de VPH16 y VPH18 en lesiones pre-neoplásicas de cuello uterino. Métodos: Se analizaron 108 muestras de raspados cervicales. El VPH se genotipificó mediante reacción de polimerasa en cadena (RPC) e hibridación no radiactiva. La integración de VPH16 y VPH18 se determinó por presencia del gen E2 mediante RPC. Resultados: VPH16 y VPH18 se detectaron en 36,1% y 12,0% de las muestras, respectivamente. El VPH16 se integró en 23,1% de los casos de VPH16, mientras que VPH18 se integró en 100% de las muestras positivas para este genotipo. Conclusiones: La integración VPH-AR es un evento temprano en la carcinogénesis cervical que ocurre en casi la mitad de las lesiones pre-neoplásicas y es más frecuente en VPH18 que en VPH16. La evaluación de la integración VPH-AR puede ser una herramienta útil para detectar el virus en la población chilena.
Background: High-risk Human Papillomaviruses (HR-HPVs) are the etiological agents of cervical cancer and its preneoplastic lesions. HPV16 and 18 are the most frequent HR-HPV genotypes detected in cervical cancer. HR-HPV genome integration into the host cell is an important event in the carcinogenic process. However, it remains uncertain which stage of cervical carcinogenesis HPV16 and 18 integration occurs in the Chilean population. Aim: The goal of this study was to evaluate HPV16 and HPV18 integration in preneoplastic lesions of the cervix. Methods: DNA was extracted from 108 cervical scrape samples with preneoplastic lesions. HPV was genotyped using PCR and non-radioactive hybridization. The integration status of HPV16 and HPV 18 was determined by evaluating the E2 gene presence through PCR. Results: HPV16 and HPV18 tested positive in 36.1% and 12.0% of samples, respectively. HPV16 was found integrated in 23.1% of HPV 16 cases, while HPV 18 in 100% of samples positive for this viral genotype. Conclusions: HR-HPV integration is an early event in cervical carcinogenesis, occurring in nearly half of preneoplastic lesions and being more frequent in HPV18 than in HPV16. The evaluation of HR-HPV integration can be utilized as a complementary tool for detecting HPV in the Chilean population.
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/geneticsABSTRACT
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.