RESUMO
Biosimilars of granulocyte colony-stimulating factor (G-CSF) have been routinely introduced into clinical practice. However, not functional genomics characterization has been performed yet in comparison with the innovator G-CSF. This study aimed to evaluate the transcriptomic changes in an in vitro model of umbilical cord blood cells (UBC) exposed to G-CSF for the identification of their modulated pathways. Umbilical cord blood cells-derived mononuclear cells (MNCs) were treated with biosimilar and innovator G-CSF for further gene expression profiling analysis using a microarray-based platform. Comparative analysis of biosimilar and innovator G-CSF gene expression signatures allowed us to identify the most commonly modulated pathways by both drugs. In brief, we observed predominantly upmodulation of transcripts related to PI3K-Akt, NF-kappaB, and tumor necrosis factor (TNF) signaling pathways as well as transcripts related to negative regulation of apoptotic process among others. In addition, hematopoietic colony-forming cell assays corroborate the G-CSF phenotypic effects over UBC-derived MNCs. In conclusion, our study suggests that G-CSF impacts UBC-derived cells through the modulation of several signaling pathways associated with cell survival, migration, and proliferation. The concordance observed between biosimilar and innovator G-CSF emphasizes their similarity in regards to their specificity and biological responses.
RESUMO
The current standard of care for locally advanced rectal cancer (RC) is neoadjuvant radio-chemotherapy (NRC) with 5-fluorouracil (5Fu) as the main drug, followed by surgery and adjuvant chemotherapy. While a group of patients will achieve a pathological complete response, a significant percentage will not respond to the treatment. The Unfolding Protein Response (UPR) pathway is generally activated in tumors and results in resistance to radio-chemotherapy. We previously showed that RHBDD2 gene is overexpressed in the advanced stages of colorectal cancer (CRC) and that it could modulate the UPR pathway. Moreover, RHBDD2 expression is induced by 5Fu. In this study, we demonstrate that the overexpression of RHBDD2 in CACO2 cell line confers resistance to 5Fu, favors cell migration, adhesion and proliferation and has a profound impact on the expression of both, the UPR genes BiP, PERK and CHOP, and on the cell adhesion genes FAK and PXN. We also determined that RHBDD2 binds to BiP protein, the master UPR regulator. Finally, we confirmed that a high expression of RHBDD2 in RC tumors after NRC treatment is associated with the development of local or distant metastases. The collected evidence positions RHBDD2 as a promising prognostic biomarker to predict the response to neoadjuvant therapy in patients with RC.
Assuntos
Resistencia a Medicamentos Antineoplásicos/genética , Regulação Neoplásica da Expressão Gênica , Proteínas de Membrana/genética , Neoplasias Retais/terapia , Resposta a Proteínas não Dobradas/efeitos dos fármacos , Antimetabólitos Antineoplásicos/farmacologia , Células CACO-2 , Adesão Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Chaperona BiP do Retículo Endoplasmático , Fluoruracila/farmacologia , Quinase 1 de Adesão Focal/genética , Quinase 1 de Adesão Focal/metabolismo , Adesões Focais/efeitos dos fármacos , Células HCT116 , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Humanos , Metástase Linfática , Proteínas de Membrana/metabolismo , Terapia Neoadjuvante/métodos , Paxilina/genética , Paxilina/metabolismo , Ligação Proteica , Neoplasias Retais/genética , Neoplasias Retais/metabolismo , Neoplasias Retais/patologia , Transdução de Sinais , Fator de Transcrição CHOP/genética , Fator de Transcrição CHOP/metabolismo , eIF-2 Quinase/genética , eIF-2 Quinase/metabolismoRESUMO
It has been established that ZFP36 (also known as Tristetraprolin or TTP) promotes mRNA degradation of proteins involved in inflammation, proliferation and tumor invasiveness. In mammary epithelial cells ZFP36 expression is induced by STAT5 activation during lactogenesis, while in breast cancer ZFP36 expression is associated with lower grade and better prognosis. Here, we show that the AP-1 transcription factor components, i.e. JUN, JUNB, FOS, FOSB, in addition to DUSP1, EGR1, NR4A1, IER2 and BTG2, behave as a conserved co-regulated group of genes whose expression is associated to ZFP36 in cancer cells. In fact, a significant down-modulation of this gene network is observed in breast, liver, lung, kidney, and thyroid carcinomas compared to their normal counterparts. In breast cancer, the normal-like and Luminal A, show the highest expression of the ZFP36 gene network among the other intrinsic subtypes and patients with low expression of these genes display poor prognosis. It is also proposed that AP-1 regulates ZFP36 expression through responsive elements detected in the promoter region of this gene. Culture assays show that AP-1 activity induces ZFP36 expression in mammary cells in response to prolactin (PRL) treatment thorough ERK1/2 activation. These results suggest that JUN, JUNB, FOS and FOSB are not only co-expressed, but would also play a relevant role in regulating ZFP36 expression in mammary epithelial cells.
Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/patologia , Mama/metabolismo , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Fator de Transcrição AP-1/metabolismo , Tristetraprolina/metabolismo , Biomarcadores Tumorais/genética , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Biologia Computacional/métodos , Feminino , Humanos , Prognóstico , Fator de Transcrição AP-1/genética , Tristetraprolina/genéticaRESUMO
Breast cancer spreading to different organs have been related to different molecules and mechanisms, but cutaneous metastasis remains unexplored. Increasing evidence showed that MUC1 and some of its carbohydrate associated antigens may be implicated in breast cancer metastasis. In this study we analyzed these tumor markers in order to identify breast cancer cutaneous metastatic profiles. A cohort of 26 primary tumors from breast cancer patients with cutaneous metastases were included; also, cutaneous and lymphatic node metastatic samples and primary tumors from breast cancer patients without metastases were analysed. Immunohistochemical (IHC) studies demonstrated that both underglycosylated MUC1 (uMUC1) and sialyl Lewis x (sLex) to be positively associated with cutaneous metastatic primary tumors (pâ¯<â¯0.05). Notably, a high percentage of tumors with cutaneous metastases were characterized as triple negative and Her2+ tumors (37.5 % and 29 %, respectively). Some discordant results were found between primary tumors and their matched cutaneous metastases. To determine if MUC1 variants may be carriers of carbohydrate antigens, subcellular fractions from a cutaneous metastatic lesion were obtained, immunoprecipitated and analyzed by Western blot. We found that the isolated uMUC1 with a molecular weight of>200â¯kDa was also the site for binding of anti-sLex MAb; in coincidence, a high correlation of positive IHC expression of both markers was observed. Our findings confirm that breast cancer cutaneous metastases were associated to highly malignant primary tumors and sustain the hypothesis that u-MUC1 and sLe x may drive breast cancer cutaneous metastases.
Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/patologia , Mucina-1/metabolismo , Antígeno Sialil Lewis X/metabolismo , Neoplasias Cutâneas/secundário , Adulto , Idoso , Biomarcadores Tumorais/análise , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
Over the last few years rhomboid genes have gained interest because of its association with cancer and neurodegenerative diseases. In previous studies, we demonstrated that human RHBDD2 is over-expressed in the advanced stages of breast and colorectal cancers, suggesting a favorable role in cell proliferation. So far little is known about the expression of RHBDD2 in other tissues and other species, and because of similarities between cancer and embryonic cells, this study focused on the evaluation of Rhbdd2 expression in embryonic and adult rat tissues. By IHC and RT-PCR, Rhbdd2 was identified in early stages of most tissues analyzed, with high expression in brain, spinal cord, kidney and embryonic skin. In adult tissues, the expression remained elevated while salivary glands became positive. Furthermore, Rhbdd2 showed a high expression in the most proliferative stages of the rat mammary gland. Indeed, similar findings were observed in the mouse mammary epithelial cell line HC11, in which Rhbdd2 resides in the Golgi apparatus, and at different stages of mouse mammary gland development. Therefore, Rhbdd2 would be implicated in embryonic and adult tissue proliferation.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Glândulas Mamárias Animais/metabolismo , Animais , Neoplasias da Mama/metabolismo , Neoplasias da Mama/fisiopatologia , Linhagem Celular , Proliferação de Células/genética , Feminino , Humanos , Imuno-Histoquímica , Glândulas Mamárias Animais/fisiopatologia , Proteínas de Membrana/genética , Camundongos , Proteínas de Neoplasias/genética , Gravidez , Ratos , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Rhomboid domain containing 2 (RHBDD2) was previously observed overexpressed and amplified in breast cancer samples. In order to identify biological pathways modulated by RHBDD2, gene expression profiles of RHBDD2 silenced breast cancer cells were analyzed using whole genome human microarray. Among the statistically significant overrepresented biological processes, we found protein metabolismwith the associated ontological terms folding , ubiquitination, and proteosomal degradationcell death, cell cycle, and oxidative phosphorylation. In addition, we performed an in silico analysis searching for RHBDD2 co-expressed genes in several human tissues. Interestingly, the functional analysis of these genes showed similar results to those obtained with the microarray data, with negative regulation of protein metabolism and oxidative phosphorylation as the most enriched gene ontology terms. These data led us to hypothesize that RHBDD2 might be involved in endoplasmic reticulum (ER) stress response. Thus, we specifically analyzed the unfolding protein response (UPR) of the ER stress process. We used a lentivirus-based approach for stable silencing of RHBDD2 mRNA in the T47D breast cancer cell line, and we examined the transcriptional consequences on UPR genes as well as the phenotypic effects on migration and proliferation processes. By employing dithiothreitol as an UPR inducer, we observed that cells with silenced RHBDD2 showed increased expression of ATF6, IRE1, PERK, CRT, BiP, ATF4, and CHOP (p <0.01). We also observed that RHBDD2 silencing inhibited colony formation and decreased cell migration. Based on these studies, we hypothesize that RHBDD2 overexpression in breast cancer could represent an adaptive phenotype to the stressful tumor microenvironment by modulating the ER stress response.
Assuntos
Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Proteínas de Neoplasias/metabolismo , Transdução de Sinais , Resposta a Proteínas não Dobradas , Neoplasias da Mama/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células , Estresse do Retículo Endoplasmático/genética , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Ontologia Genética , Inativação Gênica , Humanos , Proteínas de Membrana , Proteínas de Neoplasias/genética , Fenótipo , RNA Interferente Pequeno/metabolismo , Transdução de Sinais/genética , Transcrição Gênica , Resposta a Proteínas não Dobradas/genéticaRESUMO
Rhomboid is an evolutionary conserved and functionally diversified group of proteins composed of proteolytically active and inactive members that are involved in the modulation of multiple biological processes such as epidermal growth factor receptor signaling pathway, endoplasmic reticulum-associated degradation, cell death, and proliferation. Recently, several human rhomboid genes have been associated with the development of chronic myeloid leukemia and pituitary, colorectal, ovarian, and breast cancers. In this study, we evaluated the mRNA and protein expression profiles of rhomboid genes in cancer cell lines and breast tissue/tumor samples. In silico analysis of publicly available gene expression datasets showed that different rhomboid genes are specifically expressed according to the breast cancer intrinsic subtypes. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis showed a significant RHBDD2 mRNA overexpression in advanced breast cancer compared with normal tissue samples (p = 0.012). In addition, we found that RHBDL2 and PARL mRNA expression was associated with a low/intermediate histologic tumor grade (p = 0.024 and p = 0.015, respectively). Immunohistochemistry analysis showed a significant increase of RHBDD2 protein expression in association with breast cancer samples negative for progesterone receptor (p = 0.015). Moreover, protein expression analysis corroborated the quantitative RT-PCR results, indicating that breast primary tumors belonging to patients with a more disseminated disease expressed significantly increased levels of RHBDD2 protein compared with less disseminated tumors (p = 0.01).
Assuntos
Neoplasias da Mama/genética , Mama/metabolismo , Regulação Neoplásica da Expressão Gênica/genética , Proteínas de Neoplasias/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/genética , Mama/patologia , Neoplasias da Mama/patologia , Feminino , Perfilação da Expressão Gênica , Humanos , Células MCF-7 , Proteínas de Membrana , Análise em Microsséries , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , Transdução de Sinais/genéticaRESUMO
In previous studies, we identified rhomboid domain containing 2 (RHBDD2) gene to be markedly overexpressed in breast cancer patients that developed recurrence of the disease. In this study, we evaluated for the first time RHBDD2 gene expression in colorectal cancer (CRC). Five public available DNA microarray studies were compiled in a homogeneous dataset of 906 colorectal samples. The statistical analysis of these data showed a significant increase of RHBDD2 expression in the advanced stages of CRC (p < 0.01). We validated these findings by immunohistochemistry on 130 colorectal tissue samples; RHBDD2 protein overexpression was also observed in the advanced stages of the disease (p < 0.001). In addition, we investigated RHBDD2 expression in response to the chemotherapy agent 5-fluorouracile (5FU). We detected a significant increase of RHBDD2 mRNA and protein after 5FU treatment (20-40 µM; p < 0.001). Overall, these results showed that RHBDD2 overexpression might play a role in colorectal cancer progression.
Assuntos
Antimetabólitos Antineoplásicos/uso terapêutico , Biomarcadores Tumorais/genética , Neoplasias Colorretais/genética , Fluoruracila/uso terapêutico , Proteínas de Neoplasias/genética , Biomarcadores Tumorais/metabolismo , Western Blotting , Estudos de Casos e Controles , Colo/metabolismo , Colo/patologia , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/patologia , Perfilação da Expressão Gênica , Humanos , Técnicas Imunoenzimáticas , Proteínas de Membrana , Proteínas de Neoplasias/metabolismo , Estadiamento de Neoplasias , Análise de Sequência com Séries de Oligonucleotídeos , Prognóstico , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reto/metabolismo , Reto/patologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais CultivadasRESUMO
In this review we provide a systematic analysis of transcriptomic signatures derived from 42 breast cancer gene expression studies, in an effort to identify the most relevant breast cancer biomarkers using a meta-analysis method. Meta-data revealed a set of 117 genes that were the most commonly affected ranging from 12% to 36% of overlap among breast cancer gene expression studies. Data mining analysis of transcripts and protein-protein interactions of these commonly modulated genes indicate three functional modules significantly affected among signatures, one module related with the response to steroid hormone stimulus, and two modules related to the cell cycle. Analysis of a publicly available gene expression data showed that the obtained meta-signature is capable of predicting overall survival (P < 0.0001) and relapse-free survival (P < 0.0001) in patients with early-stage breast carcinomas. In addition, the identified meta-signature improves breast cancer patient stratification independently of traditional prognostic factors in a multivariate Cox proportional-hazards analysis.
RESUMO
The MUC1 gene is aberrantly overexpressed in approximately 90% of human breast cancers. Several studies have shown that MUC1 overexpression is due to transcriptional regulatory events. However, the importance of gene amplification as a mechanism leading to the increase of MUC1 expression in breast cancer has been poorly characterized. The aim of this study was to evaluate the role of MUC1 gene amplification and protein expression in human breast cancer development. By means of real-time quantitative polymerase chain reaction and immunohistochemical methods, 83 breast tissue samples were analyzed for MUC1 gene amplification and protein expression. This analysis showed MUC1 genomic amplification and a positive association with the histopathological group in 12% (1 out of 8) of benign lesions and 38% (23 out of 60) of primary invasive breast carcinoma samples (P = 0.004). Array-comparative genomic hybridization meta-analysis of 886 primary invasive breast carcinomas obtained from 22 studies showed MUC1 genomic gain in 43.7% (387 out of 886) of the samples. Moreover, we identified a highly statistical significant association between MUC1 gene amplification and MUC1 protein expression assessed by immunohistochemistry and Western blot test (P < 0.0001). In conclusion, this study demonstrated that MUC1 copy number increases from normal breast tissue to primary invasive breast carcinomas in correlation with MUC1 protein expression.
Assuntos
Neoplasias da Mama/genética , Carcinoma Ductal de Mama/genética , Amplificação de Genes , Mucina-1/genética , Western Blotting , Neoplasias da Mama/metabolismo , Carcinoma Ductal de Mama/metabolismo , Linhagem Celular Tumoral , Feminino , Humanos , Imuno-Histoquímica , Mucina-1/biossíntese , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
In the course of breast cancer global gene expression studies, we identified an uncharacterized gene known as RHBDD2 (Rhomboid domain containing 2) to be markedly over-expressed in primary tumors from patients with recurrent disease. In this study, we identified RHBDD2 mRNA and protein expression significantly elevated in breast carcinomas compared with normal breast samples as analyzed by SAGE (n=46) and immunohistochemistry (n=213). Interestingly, specimens displaying RHBDD2 over-expression were predominantly advanced stage III breast carcinomas (p=0.001). Western-blot, RT-PCR and cDNA sequencing analyses allowed us to identify two RHBDD2 alternatively spliced mRNA isoforms expressed in breast cancer cell lines. We further investigated the occurrence and frequency of gene amplification and over-expression affecting RHBDD2 in 131 breast samples. RHBDD2 gene amplification was detected in 21% of 98 invasive breast carcinomas analyzed. However, no RHBDD2 amplification was detected in normal breast tissues (n=17) or breast benign lesions (n=16) (p=0.014). Interestingly, siRNA-mediated silencing of RHBDD2 expression results in a decrease of MCF7 breast cancer cells proliferation compared with the corresponding controls (p=0.001). In addition, analysis of publicly available gene expression data showed a strong association between high RHBDD2 expression and decreased overall survival (p=0.0023), relapse-free survival (p=0.0013), and metastasis-free interval (p=0.006) in patients with primary ER-negative breast carcinomas. In conclusion, our findings suggest that RHBDD2 over-expression behaves as an indicator of poor prognosis and may play a role facilitating breast cancer progression.
Assuntos
Neoplasias da Mama/enzimologia , Neoplasias da Mama/genética , Regulação Neoplásica da Expressão Gênica , Proteínas de Neoplasias/genética , Serina Endopeptidases/genética , Processamento Alternativo/genética , Sequência de Bases , Biomarcadores Tumorais , Mama/patologia , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células , Análise Mutacional de DNA , Células Epiteliais/enzimologia , Feminino , Seguimentos , Perfilação da Expressão Gênica , Técnicas de Silenciamento de Genes , Inativação Gênica , Humanos , Imuno-Histoquímica , Isoenzimas/genética , Isoenzimas/metabolismo , Proteínas de Membrana , Dados de Sequência Molecular , Proteínas de Neoplasias/metabolismo , Prognóstico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Serina Endopeptidases/metabolismoRESUMO
OBJECTIVE: The aim of the present work was to evaluate the presence of human papillomavirus genotypes in malignant and normal mucosa of the colon and rectum in order to determine if a relationship exists between HPV infection and colon neoplasms. MATERIALS AND METHODS: Thirty normal colon tissues and 54 sporadic adenocarcinomas were screened for HPV positivity using nested-PCR. Detection of viral types 6, 11, 16, 18, 33, 34 and 51 was performed by the LIS-SSCP (Low Ionic Strength-Single Strand Conformational Polymorphism) procedure. RESULTS: Significant differences in high risk HPV infection were found between normal samples and adenocarcinomas (P < 0.001). Among the cases, an inverse association between HPV infection and Dukes staging was also found (P = 0.020). Finally, there was no significant association between HPV and some classical clinicopathological features, although a gradient of infection form rectum to cecum was evident. CONCLUSION: The present study demonstrates that HPV may infect the glandular mucosa of the colon and suggests a possible association between HPV and colorectal cancer.
Assuntos
Adenocarcinoma/virologia , Neoplasias do Colo/virologia , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , Neoplasias Retais/virologia , Adenocarcinoma/genética , Idoso , Idoso de 80 Anos ou mais , Distribuição de Qui-Quadrado , Neoplasias do Colo/genética , DNA Viral/análise , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Papillomaviridae/genética , Infecções por Papillomavirus/genética , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples , Neoplasias Retais/genéticaRESUMO
Cattle hypocuprosis is the second most widespread mineral deficiency affecting grazing cattle. The consequences of hypocuprosis include a failure of copper metalloenzymes, many of which form part of the antioxidant defence system. This work focuses on the association between copper (Cu) plasma concentration and DNA damage in Aberdeen Angus cattle. Two-hundred and ninety-nine heparinized blood samples from 2-year-old Aberdeen Angus cows were obtained from different farms located in the Salado River basin, Argentina. Plasma copper level analysis was carried out in whole samples, while cytogenetic analysis and single cell gel electrophoresis assay (comet assay) were carried out in 82 and 217 samples, respectively. Cytogenetic analysis showed a significant increase in the frequency of abnormal metaphases in moderate/severe hypocupremic groups (groups B and C) in relation to the normocupremic group (group A) (4.5 and 1.5 abnormal metaphases/100 cells, respectively, P < 0.01). The Spearman correlation test showed a negative association between cupremic values and the yield of chromosomal aberrations (r = -0.708, P < 0.0001). In the comet assay greater migration was observed in cells from the hypocupremic group, from a median of 54 in the severe hypocupremic group to 31 in the normocupremic group (P < 0.01). Accordingly, the Spearman correlation test showed a significant positive relationship between copper levels and cells without DNA migration and a significant negative relationship between copper levels and cells with a tiny tail (P < 0.0001 in both cases). The results obtained show that hypocupremia in cattle is associated with an increase in the frequency of chromosomal aberrations as well as in DNA migration as assessed by the comet assay. Whereas the comet assay could differentiate copper plasma level groups, chromosomal aberrations only detected differences between normal and hypocupremic animals. The increase of DNA damage found in hypocupremic animals could be explained by higher oxidative stress suffered by these animals.
Assuntos
Doenças dos Bovinos/etiologia , Cobre/deficiência , Dano ao DNA , Animais , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/genética , Aberrações Cromossômicas , Ensaio Cometa/veterinária , Cobre/sangue , Cobre/metabolismoRESUMO
Genital infection with human papillomavirus (HPV) is one of the most common sexually transmitted viral diseases. High risk HPV are now considered the main etiologic agent of cancer of the uterine cervix and their high-grade precursor lesions. The aim of the present study was to investigate the endemic HPV-genotype spectrum in a population of women from the city of La Plata, Argentina. With this purpose, 718 cervical scrapes or biopsies corresponding to 152 normal samples (Pap I/II), 84 samples classified as atypical squamous cells of undetermined significance (ASCUS), 100 condyloma, 279 low-grade squamous intraepithelial lesions (LGSIL), 82 high-grade squamous intraepithelial lesions (HGSIL), and 21 squamous cell carcinomas (SCC) were studied. The detection of HPV-DNA was performed by nested polymerase chain reaction, using My 09/11 and Gp 05/06. The viral genotypes were analyzed by single-stranded conformation polymorphisms, employing low ionic strength solution (LIS-SSCP). The overall prevalence of HPV infection was 75% in the analyzed population, with a frequency of 46% for normal cervix, 69% for ASCUS, 86% for condyloma, 80% for LGSIL, 98% for HGSIL and 100% for SCC. The most prevalent viral types were HPV 16 (35%), followed by HPV 6/11 (27% each one), HPV 33 (6%) and HPV 18 (5%). HPV 16 was the most prevalent viral type among women with LGSIL, HGSIL and SCC, representing 33%, 50% and 67% of the genital infections, respectively. HPV 6 and 11 were the most frequent viral types among samples classified as Pap I/II, ASCUS and condyloma. Women between 21 and 30 year old showed the highest prevalence of HPV positivity, compraising the 32.2% of total infections.
Assuntos
Carcinoma de Células Escamosas/virologia , Papillomaviridae/classificação , Infecções por Papillomavirus/virologia , Infecções Tumorais por Vírus/virologia , Neoplasias do Colo do Útero/virologia , Cervicite Uterina/virologia , Adolescente , Adulto , Idoso , Argentina/epidemiologia , Carcinoma de Células Escamosas/epidemiologia , Condiloma Acuminado/epidemiologia , Condiloma Acuminado/virologia , DNA Viral/genética , DNA Viral/isolamento & purificação , Feminino , Genótipo , Humanos , Pessoa de Meia-Idade , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/epidemiologia , Prevalência , Infecções Tumorais por Vírus/epidemiologia , População Urbana , Neoplasias do Colo do Útero/epidemiologia , Cervicite Uterina/epidemiologia , Displasia do Colo do Útero/epidemiologia , Displasia do Colo do Útero/virologiaRESUMO
La infección genital por el virus papiloma humano (VPH) es la enfermedad de transmisión sexual de tipo viral más común en el mundo. Loas tipos virales de alto riesgo son considerados los agentes etiológicos del cáncer de cuello uterino. El objetivo de este estudio fue analizar los genotipos del VPH en un grupo de mujeres de la ciudad de La Plata, Argentina. Se estudiaron 718 hisopados y/o biopsias cervicales correspondientes a 152 muestras normales (Pap I/II), 84 muestras clasificadas como con atipías de significado incierto (ASCUS), 100 condilomas, 279 lesiones intraepiteliales de bajo grado (LGSIL), 82 lesiones intraepiteliales de alto grado (HGSIL) y 21 carcinomas de células escamosas (SCC). La detección del genoma viral se realizó por medio de la reacción en cadena de la polimerasa (PCR), utilizando un protocolo anidado con los cebadores My 09/11 y Gp 05/06. La genotipificación del VPH se realizó por medio de la técnica de análisis de polimorfismos en la conformación de cadenas simples, utilizando solución de siembra de baja fuerza iónica (LIS-SSCP). La prevalencia general de la infección fue 75 por ciento, con 46 por ciento de muestras ADN-VPH positivas para el grupo Pap I/II, 69 por ciento para las clasificadas como ASCUS, 86 por ciento para los condilomas, 80 por ciento para los LGSIL, 98 por ciento para los HGSIL y 100 por ciento para los carcinomas de células escamosas. Los tipos virales más prevalentes fueron VPH 16 (35 por ciento), VPH 6/11 (27 por ciento cada uno), VPH 33 (6 por ciento) y VPH 18 (5 por ciento). En el grupo de mujeres con LGSIL, HGSIL y SCC el tipo viral más prevalente fue VPH 16, representando el 33 por ciento, 50 por ciento y 67 por ciento de las infecciones, respectivamente. En las mujeres con Pap I/II, ASCUS y condilomas los tipos virales más frecuentes fueron VPH 6 y 11. El grupo etario con mayor prevalencia de VPH fue el comprendido por mujeres de 21 a 30 años, acumulando el 32,2 por ciento de las infecciones totales.
Assuntos
Humanos , Argentina , Genoma Viral , Papiloma , Reação em Cadeia da PolimeraseRESUMO
La infección genital por el virus papiloma humano (VPH) es la enfermedad de transmisión sexual de tipo viral más común en el mundo. Loas tipos virales de alto riesgo son considerados los agentes etiológicos del cáncer de cuello uterino. El objetivo de este estudio fue analizar los genotipos del VPH en un grupo de mujeres de la ciudad de La Plata, Argentina. Se estudiaron 718 hisopados y/o biopsias cervicales correspondientes a 152 muestras normales (Pap I/II), 84 muestras clasificadas como con atipías de significado incierto (ASCUS), 100 condilomas, 279 lesiones intraepiteliales de bajo grado (LGSIL), 82 lesiones intraepiteliales de alto grado (HGSIL) y 21 carcinomas de células escamosas (SCC). La detección del genoma viral se realizó por medio de la reacción en cadena de la polimerasa (PCR), utilizando un protocolo anidado con los cebadores My 09/11 y Gp 05/06. La genotipificación del VPH se realizó por medio de la técnica de análisis de polimorfismos en la conformación de cadenas simples, utilizando solución de siembra de baja fuerza iónica (LIS-SSCP). La prevalencia general de la infección fue 75 por ciento, con 46 por ciento de muestras ADN-VPH positivas para el grupo Pap I/II, 69 por ciento para las clasificadas como ASCUS, 86 por ciento para los condilomas, 80 por ciento para los LGSIL, 98 por ciento para los HGSIL y 100 por ciento para los carcinomas de células escamosas. Los tipos virales más prevalentes fueron VPH 16 (35 por ciento), VPH 6/11 (27 por ciento cada uno), VPH 33 (6 por ciento) y VPH 18 (5 por ciento). En el grupo de mujeres con LGSIL, HGSIL y SCC el tipo viral más prevalente fue VPH 16, representando el 33 por ciento, 50 por ciento y 67 por ciento de las infecciones, respectivamente. En las mujeres con Pap I/II, ASCUS y condilomas los tipos virales más frecuentes fueron VPH 6 y 11. El grupo etario con mayor prevalencia de VPH fue el comprendido por mujeres de 21 a 30 años, acumulando el 32,2 por ciento de las infecciones totales. (AU)
Assuntos
Humanos , Papiloma , Reação em Cadeia da Polimerase , Genoma Viral , ArgentinaRESUMO
The aim of the present study was to evaluate the relationship between viral type and copy number of human papillomavirus (HPV) with respect to the grade of cervical disease, and also to identify the existence of an HPV type-dependent viral load effect. DNA from 275 exocervical specimens, previously evaluated for histologic diagnosis, were evaluated for HPV presence, HPV type, and viral load. Viral load determination was performed using the low stringency PCR method (LS-PCR). Significant differences were found between the samples infected with HPV16 with respect to the samples infected with other 'high-risk' viral types (HPV -18, -31, -33 or -51) and 'low-risk' types (P < 0.05). However, highly significant differences were found between the viral loads observed in the high-grade squamous intraepithelial lesions group and normal epithelium (OR = 8.53) or the low grade ones (OR = 3.10). Moreover, a high viral load was detected in the condyloma acuminatum group compared to the normal epithelia samples (p< 0.05). This work confirms the genotype-specific association of viral load to the presence of HPV16. Also, a trend to higher viral loads could be seen in the more compromised cervical lesions. An unexpected level of viral particles appeared associated to the condylomas. This fact could be explained by a productive infection with high levels of viral replication.
Assuntos
Papillomaviridae/isolamento & purificação , Displasia do Colo do Útero/virologia , Neoplasias do Colo do Útero/virologia , Adulto , DNA Viral/análise , Feminino , Humanos , Papillomaviridae/classificação , Infecções por Papillomavirus/patologia , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase , Infecções Tumorais por Vírus/patologia , Infecções Tumorais por Vírus/virologia , Neoplasias do Colo do Útero/patologia , Carga Viral , Displasia do Colo do Útero/patologiaRESUMO
BACKGROUND: Helicobacter pylori infection is presumed to be the major causal agent of chronic active gastritis in humans. The persistent infection with this pathogen would be an important factor in the pathogenesis of peptic ulcer and also gastric cancer. METHODS: We investigated relationship between H. pylori characteristics in 42 patients with normal mucosa or gastritis with minor changes and 40 patients with mild and severe gastritis. Detection and typing of vacA and cagA genes were performed using a polymerase chain reaction method. RESULTS: The analysis of vacA prevalence and the type (S1 or S2) showed non-significant differences between the two groups studied (p > 0.05). However, cagA analysis showed highly significant differences between the groups classified as normal tissue-weak gastritis and mild-severe gastritis (p < 0.0001; OR = 8.4; CI = 3.1-22.8). CONCLUSIONS: cagA status is associated to the grade of gastritis, finding higher frequencies of H. pylori cagA+ in the moderate-severe gastritis group. These highly significant differences could make cagA status a genetic marker for disease progress.
Assuntos
Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Gastrite/microbiologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/genética , Reação em Cadeia da Polimerase , Adolescente , Adulto , Idoso , Antígenos de Bactérias/análise , Proteínas de Bactérias/análise , Proteínas de Bactérias/isolamento & purificação , Dispepsia/microbiologia , Dispepsia/patologia , Feminino , Gastrite/patologia , Marcadores Genéticos , Genótipo , Humanos , Mucosa Intestinal/microbiologia , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
BACKGROUND: Helicobacter pylori infection is presumed to be the major causal agent of chronic active gastritis in humans. The persistent infection with this pathogen would be an important factor in the pathogenesis of peptic ulcer and also gastric cancer. METHODS: We investigated relationship between H. pylori characteristics in 42 patients with normal mucosa or gastritis with minor changes and 40 patients with mild and severe gastritis. Detection and typing of vacA and cagA genes were performed using a polymerase chain reaction method. RESULTS: The analysis of vacA prevalence and the type (S1 or S2) showed non-significant differences between the two groups studied (p > 0.05). However, cagA analysis showed highly significant differences between the groups classified as normal tissue-weak gastritis and mild-severe gastritis (p < 0.0001; OR = 8.4; CI = 3.1-22.8). CONCLUSIONS: cagA status is associated to the grade of gastritis, finding higher frequencies of H. pylori cagA+ in the moderate-severe gastritis group. These highly significant differences could make cagA status a genetic marker for disease progress.
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Antígenos de Bactérias/genética , Gastrite/microbiologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/genética , Reação em Cadeia da Polimerase , Antígenos de Bactérias/análise , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Técnicas de Tipagem Bacteriana/métodos , DNA Bacteriano/análise , Dispepsia/microbiologia , Dispepsia/patologia , Mucosa Gástrica/microbiologia , Gastrite/patologia , Marcadores Genéticos , Genótipo , Helicobacter pylori/classificação , Helicobacter pylori/patogenicidade , Técnicas de Amplificação de Ácido Nucleico , Estudos Retrospectivos , VirulênciaRESUMO
Genital infection with human papillomavirus (HPV) is one of the most common sexually transmitted viral diseases. High risk HPV are now considered the main etiologic agent of cancer of the uterine cervix and their high-grade precursor lesions. The aim of the present study was to investigate the endemic HPV-genotype spectrum in a population of women from the city of La Plata, Argentina. With this purpose, 718 cervical scrapes or biopsies corresponding to 152 normal samples (Pap I/II), 84 samples classified as atypical squamous cells of undetermined significance (ASCUS), 100 condyloma, 279 low-grade squamous intraepithelial lesions (LGSIL), 82 high-grade squamous intraepithelial lesions (HGSIL), and 21 squamous cell carcinomas (SCC) were studied. The detection of HPV-DNA was performed by nested polymerase chain reaction, using My 09/11 and Gp 05/06. The viral genotypes were analyzed by single-stranded conformation polymorphisms, employing low ionic strength solution (LIS-SSCP). The overall prevalence of HPV infection was 75
in the analyzed population, with a frequency of 46
for normal cervix, 69
for ASCUS, 86
for condyloma, 80
for LGSIL, 98
for HGSIL and 100
for SCC. The most prevalent viral types were HPV 16 (35
), followed by HPV 6/11 (27
each one), HPV 33 (6
) and HPV 18 (5
). HPV 16 was the most prevalent viral type among women with LGSIL, HGSIL and SCC, representing 33
, 50
and 67
of the genital infections, respectively. HPV 6 and 11 were the most frequent viral types among samples classified as Pap I/II, ASCUS and condyloma. Women between 21 and 30 year old showed the highest prevalence of HPV positivity, compraising the 32.2
of total infections.
