RESUMO
Metastatic progression and tumor evolution complicates the clinical management of cancer patients. Circulating tumor cell (CTC) characterization is a growing discipline that aims to elucidate tumor metastasis and evolution processes. CTCs offer the clinical potential to monitor cancer patients for therapy response, disease relapse, and screen 'at risk' groups for the onset of malignancy. However, such clinical utility is currently limited to breast, prostate, and colorectal cancer patients. Further understanding of the basic CTC biology of other malignancies is required to progress them towards clinical utility. Unfortunately, such basic clinical research is often limited by restrictive characterization methods and high-cost barrier to entry for CTC isolation and imaging infrastructure. As experimental clinical results on applications of CTC are accumulating, it is becoming clear that a two-tier system of CTC isolation and characterization is required. The first tier is to facilitate basic research into CTC characterization. This basic research then informs a second tier specialised in clinical prognostic and diagnostic testing. This study presented in this manuscript describes the development and application of a low-cost, CTC isolation and characterization pipeline; CTC-5. This approach uses an established 'isolation by size' approach (ScreenCell Cyto) and combines histochemical morphology stains and multiparametric immunofluorescence on the same isolated CTCs. This enables capture and characterization of CTCs independent of biomarker-based pre-selection and accommodates both single CTCs and clusters of CTCs. Additionally, the developed open-source software is provided to facilitate the synchronization of microscopy data from multiple sources (https://github.com/CTC5/). This enables high parameter histochemical and immunofluorescent analysis of CTCs with existing microscopy infrastructure without investment in CTC specific imaging hardware. Our approach confirmed by the number of successful tests represents a potential major advance towards highly accessible low-cost technology aiming at the basic research tier of CTC isolation and characterization. The biomarker independent approach facilitates closing the gap between malignancies with poorly, and well-defined CTC phenotypes. As is currently the case for some of the most commonly occurring breast, prostate and colorectal cancers, such advances will ultimately benefit the patient, as early detection of relapse or onset of malignancy strongly correlates with their prognosis.
RESUMO
Neonatal encephalopathy (NE) is characterized by altered neurological function in term infants and inflammation plays an important pathophysiological role. Inflammatory cytokines interleukin (IL)-1ß, IL-1ra and IL-18 are activated by the nucleotide-binding and oligomerization domain (NOD)-, leucine-rich repeat domain (LRR)- and NOD-like receptor protein 3 (NLRP3) inflammasome; furthermore, we aimed to examine the role of the inflammasome multiprotein complex involved in proinflammatory responses from the newborn period to childhood in NE. Cytokine concentrations were measured by multiplex enzyme-linked immunosorbent assay (ELISA) in neonates and children with NE in the absence or presence of lipopolysaccharide (LPS) endotoxin. We then investigated expression of the NLRP3 inflammasome genes, NLRP3, IL-1ß and ASC by polymerase chain reaction (PCR). Serum samples from 40 NE patients at days 1 and 3 of the first week of life and in 37 patients at age 4-7 years were analysed. An increase in serum IL-1ra and IL-18 in neonates with NE on days 1 and 3 was observed compared to neonatal controls. IL-1ra in NE was decreased to normal levels at school age, whereas serum IL-18 in NE was even higher at school age compared to school age controls and NE in the first week of life. Percentage of LPS response was higher in newborns compared to school-age NE. NLRP3 and IL-1ß gene expression were up-regulated in the presence of LPS in NE neonates and NLRP3 gene expression remained up-regulated at school age in NE patients compared to controls. Increased inflammasome activation in the first day of life in NE persists in childhood, and may increase the window for therapeutic intervention.
Assuntos
Encefalopatias/imunologia , Inflamassomos/imunologia , Inflamação/imunologia , Criança , Pré-Escolar , Citocinas/imunologia , Feminino , Humanos , Recém-Nascido , Interleucina-1beta/imunologia , Lipopolissacarídeos/imunologia , Masculino , Proteína 3 que Contém Domínio de Pirina da Família NLR/imunologia , Regulação para Cima/imunologiaRESUMO
OBJECTIVE: To estimate prevalence of post-colposcopy physical after-effects and investigate associations between these and subsequent psychological distress. DESIGN: Longitudinal survey. SETTING: Two hospital-based colposcopy clinics. POPULATION: Women with abnormal cytology who underwent colposcopy (±related procedures). METHODS: Questionnaires were mailed to women 4, 8 and 12 months post-colposcopy. Details of physical after-effects (pain, bleeding and discharge) experienced post-colposcopy were collected at 4 months. Colposcopy-specific distress was measured using the Process Outcome-Specific Measure at all time-points. Linear mixed-effects regression was used to identify associations between physical after-effects and distress over 12 months, adjusting for socio-demographic and clinical variables. MAIN OUTCOME MEASURES: Prevalence of post-colposcopy physical after-effects. Associations between the presence of any physical after-effects, awareness of after-effects, and number of after-effects and distress. RESULTS: Five-hundred and eighty-four women were recruited (response rate = 73, 59 and 52% at 4, 8 and 12 months, respectively). Eighty-two percent of women reported one or more physical after-effect(s). Multiple physical after-effects were common (two after-effects = 25%; three after-effects = 25%). Psychological distress scores declined significantly over time. In adjusted analyses, women who experienced all three physical after-effects had on average a 4.58 (95% CI: 1.10-8.05) higher distress scored than those who experienced no after-effects. Women who were unaware of the possibility of experiencing after-effects scored significantly higher for distress during follow-up. CONCLUSIONS: The prevalence of physical after-effects of colposcopy and related procedures is high. The novel findings of inter-relationships between awareness of the possibility of after-effects and experiencing multiple after-effects, and post-colposcopy distress may be relevant to the development of interventions to alleviate post-colposcopy distress. TWEETABLE ABSTRACT: Experiencing multiple physical after-effects of colposcopy is associated with psychological distress.
Assuntos
Colposcopia/efeitos adversos , Hemorragia/etiologia , Dor Processual/etiologia , Estresse Psicológico/etiologia , Doenças Vaginais/etiologia , Adulto , Colposcopia/psicologia , Feminino , Hemorragia/diagnóstico , Hemorragia/epidemiologia , Hemorragia/psicologia , Humanos , Modelos Lineares , Estudos Longitudinais , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde , Dor Processual/diagnóstico , Dor Processual/epidemiologia , Dor Processual/psicologia , Prevalência , Estresse Psicológico/diagnóstico , Estresse Psicológico/epidemiologia , Doenças Vaginais/diagnóstico , Doenças Vaginais/epidemiologia , Doenças Vaginais/psicologiaRESUMO
INTRODUCTION: The close relationship between coagulation, thrombosis and cancer has long been established. Gynaecological cancers, in particular ovarian cancers, carry a high risk of thrombosis but coagulation activation is also thought to play a role in tumorigenesis and metastasis. In experimental animal models of metastasis, mice with a genetic procoagulant phenotype are prone to develop metastasis and anticoagulant therapy dramatically reduces pulmonary metastasis in these models. The aPC pathway is a key natural anticoagulant pathway, in addition to its role in venous thrombosis, dysregulation of this pathway is also thought to play a role in the pathogenesis of some cancers. No data exists in ovarian and endometrial cancers. AIM: The aim of this study is to determine the expression of key proteins of the activated protein C pathway in endometrial and ovarian malignant tumours compared to benign tumours and to assess their role in patient survival. MATERIALS AND METHODS: RNA was extracted from 78 (54 malignant and 24 benign) fresh frozen ovarian and endometrial tumours samples. Tumour biopsies were mRNA expression of endothelial protein C receptor (EPCR), protein S (PS), protein C (PC), thrombomodulin (TM), Factor V (FV) and VIII (FVIII) and PAR-1 and PAR-2 was measured using TaqMan Low Density Arrays. mRNA fold change relative to benign expression was determined using the 2 -delta delta Ct method with 18s as internal standard. All patients gave full and informed consent and the study had the approval of the hospital ethics committee. Total cell protein was extracted from ovarian tumour tissue. Enzyme-linked immunosorbent assay (ELISA) was used to measure protein plasma expression RESULTS: EPCR (P<0.001), protein S (P<0.0001) and Factor VIII (P<0.003) mRNA expression was significantly downregulated in malignant tumours compared with benign. Factor V and PAR-2 were significantly upregulated (P<0.001; P<0.004). Protein C was not consistently expressed. Reduced EPCR and TM protein expression was also observed in malignant tumours with increased plasma levels of Factor V. Reduced protein S and increased FV were associated with decreased survival. Plasma levels of Factor V were related to grade in the endometrial cancer group. PAR-2 mRNA expression was increased in ovarian tumours (P<0.001) however PAR-1 expression remained unchanged. CONCLUSIONS: Our results show reduced expression of key proteins associated with activation of protein C combined with increased expression in FV in gynaecological malignancies. These changes may contribute to local thrombin production and tumour progression and metastasis. Further work is required to determine the precise mechanisms involved.
RESUMO
OBJECTIVE: To investigate human papillomavirus (HPV) DNA testing and p16/Ki-67 staining for detecting cervical intraepithelial grade 2 or worse (CIN2+) and CIN3 in women referred to colposcopy with minor abnormal cervical cytology low-grade squamous intraepithelial lesions (LSIL) and atypical squamous cells of undermined significance (ASC-US). The clinical performance of both tests was evaluated as stand-alone tests and combined, for detection CIN2+ and CIN3 over 2 years. METHODS: ThinPrep(®) liquid-based cytology (LBC) specimens were collected from 1349 women with repeat LSIL or ASC-US. HPV DNA was performed using Hybrid Capture. Where adequate material remained (n = 471), p16/Ki-67 overexpression was assessed. Clinical performance for detection of histologically diagnosed CIN2+ and CIN3 was calculated. RESULTS: Approximately 62.2% of the population were positive for HPV DNA, and 30.4% were positive for p16/Ki-67. p16/Ki-67 showed no significant difference in positivity between LSIL and ASC-US referrals (34.3% versus 28.6%; P = 0.189). Women under 30 years had a higher rate of p16/Ki-67 compared to those over 30 years (36.0% versus 26.6%; P = 0.029). Overall HPV DNA testing produced a high sensitivity for detection of CIN3 of 95.8% compared to 79.2% for p16/Ki-67. In contrast, p16/Ki-67 expression offered a higher specificity, 75.2% versus 40.4% for detection of CIN3. Combining p16/Ki-67 with HPV DNA improved the accuracy in distinguishing between CIN3 and Assuntos
Inibidor p16 de Quinase Dependente de Ciclina/biossíntese
, Antígeno Ki-67/biossíntese
, Lesões Intraepiteliais Escamosas Cervicais/diagnóstico
, Displasia do Colo do Útero/diagnóstico
, Adulto
, Células Escamosas Atípicas do Colo do Útero/patologia
, Colposcopia
, Inibidor p16 de Quinase Dependente de Ciclina/genética
, Citodiagnóstico
, Feminino
, Regulação Neoplásica da Expressão Gênica
, Humanos
, Antígeno Ki-67/genética
, Pessoa de Meia-Idade
, Gradação de Tumores
, Papillomaviridae/isolamento & purificação
, Papillomaviridae/patogenicidade
, Infecções por Papillomavirus/diagnóstico
, Infecções por Papillomavirus/genética
, Infecções por Papillomavirus/patologia
, Infecções por Papillomavirus/virologia
, Gravidez
, Lesões Intraepiteliais Escamosas Cervicais/genética
, Lesões Intraepiteliais Escamosas Cervicais/patologia
, Lesões Intraepiteliais Escamosas Cervicais/virologia
, Esfregaço Vaginal
, Adulto Jovem
, Displasia do Colo do Útero/genética
, Displasia do Colo do Útero/patologia
, Displasia do Colo do Útero/virologia
RESUMO
BACKGROUND: Although colposcopy is the leading follow-up option for women with abnormal cervical cytology, little is known about its psychological consequences. OBJECTIVES: We performed a systematic review to examine: (1) what, if any, are the adverse psychological outcomes following colposcopy and related procedures; (2) what are the predictors of adverse psychological outcomes post-colposcopy; and (3) what happens to these outcomes over time. SEARCH STRATEGY: Five electronic databases (PubMed, PsychINFO, CINAHL, Web of Science, Scopus) were searched for studies published in English between January 1986 and February 2014. SELECTION CRITERIA: Eligible studies assessed psychological wellbeing at one or more time-points post-colposcopy. DATA COLLECTION AND ANALYSIS: Two reviewers independently screened titles and abstracts. Full texts of potentially eligible papers were reviewed. Data were abstracted from, and a quality appraisal performed of, eligible papers. MAIN RESULTS: Twenty-three papers reporting 16 studies were eligible. Colposcopy and related procedures can lead to adverse psychological outcomes, particularly anxiety. Ten studies investigated predictors of adverse psychological outcomes; management type and treatment had no impact on this. Seven studies investigated temporal trends in psychological outcomes post-colposcopy; findings were mixed, especially in relation to anxiety and distress. Studies were methodologically heterogeneous. CONCLUSIONS: Follow-up investigations and procedures for abnormal cervical cytology can cause adverse psychological outcomes among women. However, little is known about the predictors of these outcomes or how long they persist. There is a need for a more standardised approach to the examination of the psychological impact of colposcopy, especially longer-term outcomes. TWEETABLE ABSTRACT: Follow-up investigations for abnormal cervical cytology can cause adverse psychological outcome among women.
Assuntos
Ansiedade/etiologia , Colposcopia/psicologia , Detecção Precoce de Câncer/psicologia , Estresse Psicológico/etiologia , Displasia do Colo do Útero/psicologia , Neoplasias do Colo do Útero/psicologia , Detecção Precoce de Câncer/instrumentação , Feminino , Humanos , Neoplasias do Colo do Útero/patologia , Displasia do Colo do Útero/patologiaRESUMO
OBJECTIVE: To explore emotional responses, and predictors of negative reactions, among women undergoing human papillomavirus (HPV) tests in routine clinical practice. DESIGN: Exploratory qualitative interview study. SETTING: A large busy colposcopy clinic in a Dublin hospital. SAMPLE: Twenty-seven women who had had an HPV DNA test in the previous 6 months following one or more low-grade cytology tests or treatment for cervical intraepithelial neoplasia (CIN). METHODS: In-depth semi-structured interviews were conducted. Interview transcripts were analysed using a thematic approach (Framework Analysis). MAIN OUTCOME MEASURES: Women's emotional responses and predictors of negative emotional reactions. RESULTS: For most women, having a test for high-risk HPV types generated little negative or positive emotional impact. Adverse emotional responses related to HPV infection rather than testing. Factors that influenced whether women experienced negative emotional responses were: concerns over abnormal cytology or diagnosis of CIN; HPV knowledge; awareness of HPV being sexually transmitted; awareness of HPV prevalence; and HPV information needs. Women's concerns about abnormal cytology/CIN dominated all other issues. CONCLUSIONS: These qualitative data suggest that in the context of follow up of abnormal cytology or treatment for CIN, the emotional impact of HPV testing may be modest: women's primary concerns at this time relate to abnormal cytology/CIN.
Assuntos
Ansiedade , Colposcopia , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/psicologia , Displasia do Colo do Útero/psicologia , Neoplasias do Colo do Útero/psicologia , Adolescente , Adulto , Colposcopia/psicologia , Aconselhamento , Emoções , Feminino , Seguimentos , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Irlanda , Programas de Rastreamento/psicologia , Pessoa de Meia-Idade , Infecções por Papillomavirus/diagnóstico , Lesões Pré-Cancerosas/psicologia , Valor Preditivo dos Testes , Pesquisa Qualitativa , Inquéritos e Questionários , Neoplasias do Colo do Útero/diagnóstico , Esfregaço Vaginal/métodos , Displasia do Colo do Útero/diagnósticoRESUMO
A Lab-On-Chip system with an instrument is presented which is capable of performing total sample preparation and automated extraction of nucleic acid from human cell samples fixed in a methanol based solution. The target application is extraction of mRNA from cervical liquid based cytology specimens for detection of transformed HPV-infections. The device accepts 3 ml of sample and performs the extraction in a disposable polymer chip of credit card size. All necessary reagents for cell lysis, washing, and elution are stored on-chip and the extraction is performed in two filter stages; one for cell pre-concentration and the other for nucleic acid capture. Tests performed using cancer cell lines and cervical liquid based cytology specimens confirm the extraction of HPV-mRNA by the system.
Assuntos
Dispositivos Lab-On-A-Chip , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/diagnóstico , RNA Viral/isolamento & purificação , Esfregaço Vaginal/instrumentação , Linhagem Celular Tumoral , Desenho de Equipamento , Feminino , Humanos , Dispositivos Lab-On-A-Chip/economia , RNA Mensageiro/isolamento & purificação , Sensibilidade e Especificidade , Esfregaço Vaginal/economiaRESUMO
RET/PTC rearrangements are initiating events in the development of a significant proportion of papillary thyroid carcinomas. Activated RET/PTC mutations are thought to be restricted to thyroid disease, but this study proposes that these events may also occur in nonthyroid tumors. A total of 57 nonthyroid papillary tumors were examined for RET/PTC rearrangements using interphase fluorescence in situ hybridization, Taqman reverse transcriptase polymerase chain reaction, and immunohistochemistry. Taqman single nucleotide polymorphism detection was used to analyze for expression of mutated BRAF T1799A. In all, 20% (3/15) of primary peritoneal carcinoma had detectable RET/PTC1 rearrangements by all 3 methodologies. A further case of similar histotype had an alternate RET/ PTC rearrangement. No RET/PTC1 rearrangements were detected in the remaining tumor cohort. All 57 tumors were homozygous for wild-type BRAF. The results indicate that RET/PTC rearrangements occur in a small subset of nonthyroid papillary tumors. These rearrangements may not be directly implicated in tumor growth; rather representing "passenger" mutations reflecting RET instability in secondary tumor subclones.
Assuntos
Biomarcadores/análise , Carcinoma Papilar/genética , Carcinoma/genética , Rearranjo Gênico , Neoplasias Peritoneais/genética , Proteínas Proto-Oncogênicas c-ret/genética , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Masculino , Pessoa de Meia-Idade , Proteínas de Fusão Oncogênica/genética , Proteínas Proto-Oncogênicas B-raf/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The most common sub-variant of papillary thyroid carcinoma (PTC) is the so-called follicular variant (FVPTC), which is a particularly problematic lesion and can be challenging from a diagnostic viewpoint even in resected lesions. Although fine needle aspiration cytology is very useful in the diagnosis of PTC, its accuracy and utility would be greatly facilitated by the development of specific markers for PTC and its common variants. We used the recently developed Applied Biosystems 1700 microarray system to interrogate a series of 11 benign thyroid lesions and conditions and 14 samples of PTC (six with classic morphology and eight with follicular variant morphology). TaqMan(R) reverse transcriptase-polymerase chain reaction was used to validate the expression portfolios of 50 selected transcripts. Our data corroborates potential biomarkers previously identified in the literature, such as LGALS3, S100A11, LYN, BAX, and cluster of differentiation 44 (CD44). However, we have also identified numerous transcripts never previously implicated in thyroid carcinogenesis, and many of which are not represented on other microarray platforms. Diminished expression of metallothioneins featured strongly among these and suggests a possible role for this family as tumour suppressors in PTC. Fifteen transcripts were significantly associated with FVPTC morphology. Surprisingly, these genes were associated with an extremely narrow repertoire of functions, including the major histocompatibility complex and cathepsin families.
Assuntos
Adenocarcinoma Folicular/genética , Adenocarcinoma Papilar/genética , Biomarcadores Tumorais/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Neoplasias da Glândula Tireoide/genética , Adenocarcinoma Folicular/metabolismo , Adenocarcinoma Folicular/patologia , Adenocarcinoma Papilar/metabolismo , Adenocarcinoma Papilar/patologia , Biomarcadores Tumorais/metabolismo , Expressão Gênica , Perfilação da Expressão Gênica , Humanos , Reação em Cadeia da Polimerase/métodos , Estudos Prospectivos , RNA Mensageiro/metabolismo , Taq Polimerase/metabolismo , Neoplasias da Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/patologia , TireoidectomiaRESUMO
BACKGROUND: The aim of this study was to quantify maternal plasma fetal DNA and total DNA in early pregnancy in intrauterine growth restriction (IUGR) or pre-eclampsia (PET). METHODS: A nested case control study was carried out in a University Teaching Hospital. Plasma samples were obtained from 1993 women before 20 weeks of gestation. Pregnancies complicated by IUGR or PET were identified and compared to controls. DNA was extracted and real-time quantitative PCR applied for the SRY and beta-actin genes. IUGR or PET groups were compared to controls using the chi(2) and Wilcoxon rank sum tests. RESULTS: SRY was detected in 86% of IUGR (31/36), 94% of PET (15/16) and 78% of controls (56/72). The median SRY was similar in women with IUGR (28 GE/mL) or PET (30.5 GE/mL) and controls (27.5 GE/mL). beta-actin was increased in the IUGR group (3975 GE/mL) compared to controls (1835 GE/mL) (p = 0.045). Cigarette consumption was greater in the IUGR group compared to controls (p = 0.004). CONCLUSIONS: Fetal DNA quantitation in maternal plasma before 20 weeks is not a useful predictor of IUGR or PET. beta-actin levels were elevated before 20 weeks in women with IUGR and may be a marker of maternal susceptibility to this condition.
Assuntos
DNA/sangue , Retardo do Crescimento Fetal/diagnóstico , Feto , Troca Materno-Fetal , Pré-Eclâmpsia/diagnóstico , Actinas/sangue , Actinas/genética , Adolescente , Adulto , Estudos de Casos e Controles , Feminino , Retardo do Crescimento Fetal/sangue , Retardo do Crescimento Fetal/genética , Genes sry/genética , Predisposição Genética para Doença , Idade Gestacional , Humanos , Masculino , Pré-Eclâmpsia/sangue , Pré-Eclâmpsia/genética , Gravidez , Diagnóstico Pré-Natal , Proteína da Região Y Determinante do Sexo/sangue , Proteína da Região Y Determinante do Sexo/genéticaRESUMO
OBJECTIVE: The aim of this study was to assess whether microvessel density (measured by CD31), vascular endothelial growth factor (VEGF) or multidrug resistance (MDR1) could determine the response to chemotherapy or act as prognostic factors in ovarian cancer. METHODS: Seventy-nine ovarian specimens were immunostained. Pearson correlation, 1-way ANOVA and chi-square were used for univariate analysis. Kaplan Meier survival curves were used, log-rank was used for univariate analysis and a Cox proportional hazards regression model was used for multivariate evaluation. Response to chemotherapy was assessed after 6 months and again after 1 year. RESULT: Quantifying VEGF proved to be a valuable independent prognostic indicator in progression-free survival (PFS) (p<0.05) and overall survival (OS) (p<0.0001). VEGF correlated with response to chemotherapy at the 6-month interval (r=0.446, p<0.001) but failed to correlate at the 1-year interval. Increased staining with CD31 was associated with decreased PFS (p<0.01) and OS (p<0.01) in univariate but not multivariate analysis. MDR1 failed to act as a prognostic marker or as a predictor of response to chemotherapy. CONCLUSION: VEGF correlates with response to chemotherapy at the 6-month but not the 12-month interval. What should our criteria be for determining sensitivity to chemotherapy? CD31, VEGF and MDR1 do play a role in some ovarian malignancies but other factors are likely to be involved and perhaps molecular profiling will determine which factors will be important for determining the response to chemotherapy.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias Ovarianas/irrigação sanguínea , Neoplasias Ovarianas/tratamento farmacológico , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Intervalo Livre de Doença , Resistencia a Medicamentos Antineoplásicos , Feminino , Humanos , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Compostos Organoplatínicos/administração & dosagem , Neoplasias Ovarianas/metabolismo , Paclitaxel/administração & dosagem , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Valor Preditivo dos Testes , Estudos Retrospectivos , Taxa de Sobrevida , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
The aim of this study was to identify amplified oncogenes in endometrial cancer using array-based comparative genomic hybridization (array CGH). Despite its prevalence, the molecular mechanisms of endometrial carcinogenesis are still poorly understood. The selected array CGH allows the simultaneous examination of 58 oncogenes commonly amplified in human cancers and is capable of achieving increased mapping resolution compared with conventional CGH. A subset of 8 specimens from a bank of 60 malignant and normal specimens was selected for array analysis to identify potential genes of interest. TaqMan polymerase chain reaction was carried out on the 60 specimens to examine if aberrations at the genomic level correlated with gene expression and to compare expression in normal and malignant samples. Oncogenes amplified in the endometrial cancers included AR, PIK3CA, MET, HRAS, NRAS, D17S1670, FGFR1, CTSB, RPS6KB1, LAMC2, MYC, PDGFRA, FGF4/FGF3, PAKI, and FGR. Three genes were examined at the messenger RNA level. AR and PIK3CA were higher in normal specimens, and MET was higher in malignant samples, suggesting a role for MET in endometrial cancer. Newer arrays examining more genes and larger sample numbers are necessary to elucidate the carcinogenic pathway in endometrial cancer.
Assuntos
DNA/genética , Neoplasias do Endométrio/genética , Regulação Neoplásica da Expressão Gênica , Genoma Humano , Análise de Sequência com Séries de Oligonucleotídeos , Oncogenes/genética , Adenocarcinoma/genética , Adenocarcinoma Papilar/genética , Adenocarcinoma Papilar/metabolismo , Adenocarcinoma Papilar/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carcinoma Adenoescamoso/genética , Carcinoma Adenoescamoso/metabolismo , Carcinoma Adenoescamoso/patologia , Estudos de Casos e Controles , Cistadenocarcinoma Seroso/genética , Cistadenocarcinoma Seroso/metabolismo , Cistadenocarcinoma Seroso/patologia , DNA de Neoplasias/genética , Neoplasias do Endométrio/metabolismo , Neoplasias do Endométrio/patologia , Endométrio/metabolismo , Feminino , Humanos , Hibridização in Situ Fluorescente , Pessoa de Meia-Idade , Hibridização de Ácido Nucleico , Células Tumorais Cultivadas , Regulação para CimaRESUMO
Classically, squamous cell carcinoma (SCC) of the head and neck is a disease of older adults, but recently there have been reports of an increasing incidence in young people. This study of patients in the Republic of Ireland compares sex distribution, sites, risk factors, stage and grade of tumour, and nodal status of 130 patients with SCC of the head and neck, 30 of whom were less than 40 years old. There was a highly significant association between age, smoking status, and site of tumour. For the first time to our knowledge in a study such as this, the preoperative haematological status of the patients was assessed, and although 15% were anaemic there was no significant difference in the occurrence of anaemia between the younger and the older patients. We think that it is possible that the biology of SCC of the head and neck in young people differs from that in older people.
Assuntos
Carcinoma de Células Escamosas/epidemiologia , Neoplasias de Cabeça e Pescoço/epidemiologia , Adolescente , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Distribuição de Qui-Quadrado , Feminino , Humanos , Irlanda/epidemiologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Distribuição por Sexo , Fumar/efeitos adversosRESUMO
AIM: To analyse and compare expression patterns of three potential biomarkers-p16(INK4A), CDC6, and MCM5-and evaluate their use as predictive biomarkers in squamous and glandular cervical preinvasive neoplasia. METHODS: Immunocytochemical analysis of p16(INK4A), MCM5, and CDC6 expression was performed on 20 normal, 38 cervical intraepithelial neoplasia 1 (CIN1), 33 CIN2, 46 CIN3, 10 squamous cell carcinoma, 19 cervical glandular intraepithelial neoplasia (cGIN), and 10 adenocarcinoma samples. Staining intensity was assessed using a 0-3 scoring system. p16(INK4A), MCM5, and CDC6 expression was also examined in ThinPrep slides exhibiting mild, moderate, and severe dyskaryosis. Human papillomavirus (HPV) was detected using a modified SYBR green assay. Fluorogenic polymerase chain reaction (PCR) and solution phase PCR were used for specific HPV typing. RESULTS: All three markers showed a linear correlation between expression and grade of dysplasia. p16(INK4A) and MCM5 protein expression was upregulated in all grades of squamous and glandular dysplasia. CDC6 protein was preferentially expressed in high grade lesions and in invasive squamous cell carcinoma. CONCLUSION: p16(INK4A) expression was closely associated with high risk HPV infection-all grades of squamous and glandular cervical lesions were immunohistochemically positive. MCM5 staining intensity was independent of high risk HPV infection, highlighting its potential as a biomarker in both HPV dependent and independent cervical dysplasia. CDC6 may be a biomarker of high grade and invasive lesions of the cervix, with limited use in low grade dysplasia. p16(INK4A) was the most reliable marker of cervical dysplasia. Combinations of dysplastic biomarkers may be useful in difficult diagnostic cases.
Assuntos
Biomarcadores Tumorais/análise , Proteínas de Ciclo Celular/análise , Inibidor p16 de Quinase Dependente de Ciclina/análise , Proteínas de Ligação a DNA/análise , Proteínas Nucleares/análise , Proteínas de Schizosaccharomyces pombe/análise , Displasia do Colo do Útero/química , Neoplasias do Colo do Útero/química , Adenocarcinoma/química , Adenocarcinoma/complicações , Adenocarcinoma/imunologia , Anticorpos Monoclonais/imunologia , Western Blotting/métodos , Carcinoma de Células Escamosas/química , Carcinoma de Células Escamosas/complicações , Carcinoma de Células Escamosas/imunologia , Proteínas de Ciclo Celular/imunologia , Proteínas de Ligação a DNA/imunologia , Feminino , Humanos , Imuno-Histoquímica/métodos , Proteínas de Neoplasias/análise , Proteínas de Neoplasias/imunologia , Proteínas Nucleares/imunologia , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/diagnóstico , Fatores de Risco , Proteínas de Schizosaccharomyces pombe/imunologia , Neoplasias do Colo do Útero/complicações , Neoplasias do Colo do Útero/imunologia , Displasia do Colo do Útero/complicações , Displasia do Colo do Útero/imunologiaRESUMO
The purpose of this study was to assess BRAF mutation rates in various thyroid tissues and to investigate if concomitant mutations with ret/PTC activation occurred in inflammatory and neoplastic lesions. To this end, we developed a novel Taqman based screening assay for the common T1799A BRAF mutation. Heterozygous T1799A mutations were detected in 13 of 34 (44%) papillary thyroid carcinomas (PTCs) tested. No such mutations were detected in the other tissue types tested. Concomitant presence of both oncogenes was reported in 5 of the 34 PTCs. A significant temporal trend was observed, with ret/PTC chimera detected for the most part before 1997 and BRAF mutations being more prevalent after 1997. The results suggest that some environmental/etiological agent(s) may have influenced the pathobiology of thyroid tumor development, among the population examined, over time.
Assuntos
Carcinoma Papilar/genética , Mutação , Proteínas de Fusão Oncogênica/genética , Proteínas Proto-Oncogênicas B-raf/genética , Neoplasias da Glândula Tireoide/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Papilar/epidemiologia , Carcinoma Papilar/patologia , Estudos de Coortes , DNA de Neoplasias/análise , Feminino , Humanos , Irlanda/epidemiologia , Masculino , Pessoa de Meia-Idade , Proteínas Tirosina Quinases , Neoplasias da Glândula Tireoide/epidemiologia , Neoplasias da Glândula Tireoide/patologia , Tireoidite Autoimune/genética , Tireoidite Autoimune/patologiaRESUMO
A wide array of immunohistochemical markers have been evaluated with respect to their specificity in staining dysplastic cervical cells in cervical biopsies and cervical cytological smears. However, there is still a significant demand for better biomarkers to identify neoplastic cervical glandular and squamous epithelial cells precisely. The CDKN2A gene, located on chromosome 9p21, encodes the tumour suppressor protein, p16INK4A, which decelerates the cell cycle by inactivating CDK4 and CDK6. The aim of this study was to compare and contrast the expression pattern of p16INK4A in benign and neoplastic glandular lesions and tubo-endometrioid metaplasia. All cases in each category displayed some p16INK4A expression. Adenocarcinoma and in situ cases showed a combination of intense nuclear and cytoplasmic staining. It was observed that all cases of tubo-endometrioid metaplasia showed occasional nuclear positivity and definite cytoplasmic staining. These findings may have important implications for the potential utility of p16INK4A as a biomarker for glandular dysplastic lesions. While p16INK4A has been demonstrated to be an excellent marker of cervical dysplasia in squamous neoplastic lesions of the cervix, it has potential pitfalls in cervical glandular lesions that may limit the utility of this biomarker in resolving the nature of suspicious glandular lesions, particularly in cytopathology.
Assuntos
Adenocarcinoma/química , Inibidor p16 de Quinase Dependente de Ciclina/análise , Lesões Pré-Cancerosas/química , Displasia do Colo do Útero/química , Neoplasias do Colo do Útero/química , Adenocarcinoma/patologia , Endométrio/química , Feminino , Humanos , Imuno-Histoquímica , Metaplasia , Lesões Pré-Cancerosas/patologia , Neoplasias do Colo do Útero/patologiaRESUMO
We investigated whether fetal DNA from the maternal blood can be used to diagnose abnormalities of chromosomes 13, 16, 18 and 21 in spontaneous miscarriage. Venous blood was obtained from 50 women with a diagnosis of spontaneous miscarriage. The evacuated products of the uterus were also sampled. DNA was extracted from both blood and tissue samples. Polymerase chain reaction (PCR) was performed using microsatellite markers for selected chromosomes. A novel technique using a capillary sequencer was employed. The electrophoretogram created was analysed using GeneScan software. The result obtained from the maternal blood samples was compared to the result obtained from uterine samples. Forty-two of the 50 (84%) cases were found to have a trisomy of one or more of the chromosomes tested on analysis of maternal blood. Compared to the overall result obtained on the uterine products of conception, the sensitivity was 97.6 and specificity 1.0. Trisomy 16 was the most common abnormality detected. These results suggest that the incidence of genetic abnormalities complicating miscarriage may be higher than suspected hitherto.