RESUMO
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.
Assuntos
Humanos , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Adulto Jovem , Lesões Pré-Cancerosas/virologia , Colo do Útero/virologia , Integração Viral/genética , Papillomaviridae/isolamento & purificação , Papillomaviridae/genética , Lesões Pré-Cancerosas/genética , DNA Viral/genética , Colo do Útero/patologia , Chile , Reação em Cadeia da Polimerase , Estudos Transversais , Papillomavirus Humano 16/isolamento & purificação , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/isolamento & purificação , Papillomavirus Humano 18/genética , Técnicas de Genotipagem , GenótipoRESUMO
RESUMEN: La Pansinusitis aguda odontogénica es un cuadro infeccioso infrecuente que afecta a todos los senos paranasales, en el cual se hace necesario un diagnóstico precoz para obtener una menor morbilidad. Un correcto diagnóstico requiere una evaluación dental exhaustiva, apoyándose con imágenes apropiadas. El presente caso clínico reporta paciente femenino de 17 años, sin enfermedades crónicas de base, diagnosticada con pansinusitis aguda odontogénica a través de examen de tomografía axial computarizada. Fue manejada de forma intrahospitalaria con un equipo multidisciplinario para su recuperación. Tratada con antibioterapia de amplio espectro vía parenteral y manejo del dolor, posterior al alta médica se realizó endodoncia del diente afectado y rehabilitación con prótesis fija unitaria.
ABSTRACT: Acute Odontogenic Pansinusitis is an infrequent infectious disease that affects all the paranasal sinuses. It requires an early diagnosis to obtain a lower morbidity. A correct diagnosis requires thorough dental evaluation, supported by appropriate images. The present clinical case reports a 17-year-old female patient, without chronic underlying diseases, with anacute Odontogenic Pansinusitis diagnosed through a computerized axial tomography scan. She was treated by a multidisciplinary team for recovery, through parenteral wide spectrum antibiotic therapy and pain management. After medical discharge, root canal of the affected tooth and rehabilitation with fixed unitary prosthesis were performed.
Assuntos
Humanos , Feminino , Adolescente , Seios Paranasais , Dente , Endodontia , Manejo da DorRESUMO
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.
Assuntos
Humanos , Feminino , Neoplasias Ovarianas/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Carboplatina/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Transcriptoma/efeitos dos fármacos , Antineoplásicos/farmacologia , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/tratamento farmacológico , Fenótipo , Transdução de Sinais , Morte Celular/efeitos dos fármacos , Morte Celular/genética , Análise de Sequência de RNA , Linhagem Celular Tumoral , Transcriptoma/genéticaRESUMO
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.
Assuntos
Feminino , Humanos , Neoplasias da Mama/patologia , Proteínas de Ciclo Celular/fisiologia , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Glicoproteínas/fisiologia , Análise de Variância , Western Blotting , Neoplasias da Mama/genética , Ciclo Celular , Linhagem Celular Tumoral , Sobrevivência Celular , Proteínas de Ciclo Celular/genética , Movimento Celular/genética , Proliferação de Células/genética , Metilação de DNA , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , Glicoproteínas/genética , Invasividade Neoplásica , Reação em Cadeia da Polimerase em Tempo Real , Estatísticas não ParamétricasRESUMO
Overexpression of Short and Raji variants of Cellular FLICE-like inhibitory protein (c-FLIP) is capable of inhibiting apoptosis, while the function of the Long isoform depends of c-FLIPL concentration in cells. The aim of this study was to determine the effects of c-FLIPL knockdown in cervical cell lines. SiHa, C-4I and C-33A cervical cancer cell lines were analyzed. c-FLIPL level expression was determined by quantitative real-time PCR and western blotting. c-FLIPL was transiently downregulated by siRNA. The effects of knockdown of c-FLIPL on cell viability, proliferation and apoptosis were assessed by comparing with scrambled siRNA-transfected cells. SiHa and C-4I c-FLIPL knockdown cells showed increased viability compared with scrambled siRNA-transfected cells (P<0.05), while C-33A cells did not show significant differences. Ki-67 and PCNA immunocytochemistry was performed to evaluate proliferation on these cervical cancer cell lines. SiHa cells with c-FLIPL knockdown showed elevated expression of Ki-67 protein compared with their scrambled counterparts (P<0.0001), while C-33A c-FLIPL knockdown cells showed a significantly lower in PCNA expression (P<0.01) compared with control. All three c-FLIP-transfected cell lines showed a higher level of apoptosis compared with their scrambled controls. Our results suggest that c-FLIPL could have effects in proliferation and apoptosis in cervical cancer cell lines.
Cuando las variantes Short y Raji de la proteína Cellular FLICE-like inhibitory protein (c-FLIP) se encuentran sobrexpresadas son capaces de inhibir la apoptosis, mientras la función de la isoforma Long (c-FLIPL), depende de la concentración de esta molécula en las células. El objetivo de este estudio fue determinar los efectos de la inhibición de c-FLIPL en líneas celulares de cáncer de cuello uterino. Para realizar el estudio fueron utilizadas SiHa, C-4I y C-33A, líneas celulares de cáncer cervical. La expresión de c-FLIPL en estas líneas fue establecida mediante PCR en tiempo real y western blot. Posteriormente la expresión de c-FLIPL fue inhibida, mediante transfeción transiente con siRNA complementario al mRNA mensajero de c.-FLIPL. Los efectos de esta inhibición en la viabilidad celular, proliferación y apoptosis fue comparada con células transfectadas con un siRNA control (scrambled). Una vez reprimido c-FLIPL, las líneas celulares SiHa y C-4I presentaron un aumento de la viabilidad celular (P<0,05). Para evaluar la proliferación celular se utilizó inmunocitoquímica de los marcadores Ki-67 y PCNA. Las células SiHa transfectadas con siRNA c-FLIPL, mostraron una elevada expresión de Ki-67 (P<0,0001), mientras que las células C-33A con c-FLIPL inhibido mostraron una menor expresión de PCNA (P<0,01). Las tres líneas celulares con c-FLIPL reprimido mostraron un mayor nivel de apoptosis que las células control. Estos resultados sugieren que c-FLIPL puede tener efectos en la proliferación y apoptosis de líneas celulares de cáncer de cuello uterino.
Assuntos
Humanos , Feminino , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologia , Linhagem Celular Tumoral , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/metabolismo , Imuno-Histoquímica , Sobrevivência Celular , Apoptose , Marcação In Situ das Extremidades Cortadas , RNA Interferente Pequeno , Proliferação de Células , Reação em Cadeia da Polimerase em Tempo RealRESUMO
El Virus Papiloma Humano (HPV por sus siglas en inglés) es una de las infecciones de transmisión sexual más frecuentes del mundo y se encuentra presente en la mayoría de los cánceres de cuello uterino. Se ha descrito su presencia en otros tipos de cáncer no ginecológicos como lo son esófago y próstata. Sin embargo, las frecuencias de HPV descritas hasta el momento para estos tipos de cáncer son muy variables, y no hay artículos donde se muestren la presencia de HPV en estas neoplasias en Chile. El objetivo de este estudio fue determinar la frecuencia de HPV en muestras de biopsias de tumores no ginecológicos y tejido inflamatorio de pacientes de la región de La Araucanía. Se extrajo DNA desde un total de 47 biopsias de pacientes con esofagitis, 25 con carcinoma escamoso esofágico, 20 con hiperplasia nodular de la próstata y 39 con adenocarcinoma prostático. Estas fueron analizadas por PCR de la región L1 del virus y posterior genotipificación por reverse line blot. Se detectó HPV en el 53,2% de las muestras de esofagitis, 48% en muestras de carcinoma escamoso esofágico, 15% en hiperplasia nodular de la próstata y un 15,4% en los casos de adenocarcinoma prostático. Siendo los más frecuentes los genotipos de HPV 16 y 18, ya sea en infecciones simples o junto con otros genotipos, en lesiones preneoplásicas y neoplásicas de los tejidos estudiados. Existe una alta frecuencia de infección por HPV en biopsias de esofagitis y tejido inflamatorio esofágico de pacientes de la región de la Araucanía. En los casos de adenocarcinoma prostático e hiperplasia nodular de la próstata se observa una baja frecuencia de HPV.
Human Papilloma Virus (HPV) is the most common sexually transmitted disease in the world and it is present in practically all cervical cancers. Its presence was described in other types of non-gynecologic cancer such as esophageal and prostate. However, HPV frequency described for these cancers is highly variable, and there are no articles describing the presence of HPV in these tumors in Chile. To determine HPV frequency in samples from biopsies of non-gynecological tumors and inflammatory tissue from patients in the Araucanía region, DNA was extracted from a total of 47 biopsies from patients with esophagitis, 25 with esophageal squamous cell carcinoma, 20 with prostate nodular hyperplasia and 39 with prostate adenocarcinoma. These were analyzed by PCR of HPV L1 region and subsequent genotyping by reverse line blot. HPV was detected in 53.2% of esophagitis samples, 48% in esophageal squamous cell carcinoma, 15% in prostatitis and 15.4% in cases of prostatic adenocarcinoma. The most frequent HPV genotypes were 16 and 18, either single or in combination with other genotype infections, in inflammatory tissue and neoplastic lesions. In patients of the Araucanía region, there is a high rate of HPV infection in biopsies obtained in esophagitis and esophageal inflammatory tissue. In cases of prostatic adenocarcinoma and prostate nodular hyperplasia a low rate of HPV was observed.
Assuntos
Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Neoplasias da Próstata/virologia , Neoplasias Esofágicas/virologia , Infecções por Papillomavirus/complicações , Papillomaviridae/isolamento & purificação , Papillomaviridae/genética , Hiperplasia Prostática/virologia , DNA Viral , Carcinoma de Células Escamosas/virologia , Adenocarcinoma/virologia , Chile , Reação em Cadeia da Polimerase , Infecções por Papillomavirus/virologia , Esofagite/virologia , GenótipoRESUMO
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.
Assuntos
Adulto , Feminino , Humanos , Gravidez , Infecções por Chlamydia/epidemiologia , Chlamydia trachomatis/isolamento & purificação , Fatores Etários , Chile/epidemiologia , Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/genética , DNA Bacteriano/análise , Prevalência , Reação em Cadeia da Polimerase em Tempo RealRESUMO
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.