RESUMO
BRAF, NRAS and TERT mutations occur in more than 2/3 of melanomas. Its detection in patient's blood, as circulating tumor DNA (ctDNA), represents a possibility for identification and monitoring of metastatic disease. We proposed to standardize a liquid biopsy platform to identify hotspot mutations in BRAF, NRAS and TERT in plasma samples from advanced melanoma patients and investigate whether it was associated to clinical outcome. Firstly, we performed digital polymerase chain reaction using tumor cell lines for validation and determination of limit of detection (LOD) of each assay and screened plasma samples from healthy individuals to determine the limit of blank (LOB). Then, we selected 19 stage III and IV patients and determined the somatic mutations status in tumor tissue and track them in patients' plasma. We established a specific and sensitive methodology with a LOD ranging from 0.13 to 0.37%, and LOB ranging from of 0 to 5.201 copies/reaction. Somatic mutations occurred in 17/19 (89%) patients, of whom seven (41%) had ctDNA detectable their paired plasma. ctDNA detection was associated with shorter progression free survival (p = 0.01). In conclusion, our data support the use of ctDNA as prognosis biomarker, suggesting that patients with detectable levels have an unfavorable outcome.
Assuntos
DNA Tumoral Circulante/sangue , Melanoma/patologia , Neoplasias Cutâneas/patologia , Adulto , Idoso , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/genética , DNA Tumoral Circulante/genética , Intervalo Livre de Doença , Feminino , GTP Fosfo-Hidrolases/genética , Humanos , Limite de Detecção , Biópsia Líquida , Masculino , Melanoma/sangue , Proteínas de Membrana/genética , Pessoa de Meia-Idade , Mutação , Reação em Cadeia da Polimerase/métodos , Prognóstico , Proteínas Proto-Oncogênicas B-raf/genética , Neoplasias Cutâneas/sangue , Telomerase/genéticaRESUMO
Characteristics of melanoma cells have been deciphered by studies carried out in two dimensional cell cultures growing as adherent monolayers on the bottom of plastic flasks. Melanoma cells can be cultured with a considerable degree of success, and, depending on the further use of the cells obtained in the culture, methodologies have to be adjusted to obtain reliable results. Although there are many melanoma continuous cell lines, in vitro 2D and 3D melanoma primary cell culture may be a more useful model to investigate interactions between cancer cells and immune system, as well as the effect of cytotoxic treatments and personalized medicine in environments more similar to the physiological conditions.Here, we described a protocol which employs many strategies to obtain primary 2D and 3D melanoma cultures as a model to study cell-cell and cell-microenvironment interactions that must be considered to properly design personalized cancer treatments, as well as for testing novel anticancer drugs and drug delivery vehicles.
Assuntos
Antineoplásicos/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Melanoma/patologia , Cultura Primária de Células/métodos , Neoplasias Cutâneas/patologia , Antineoplásicos/uso terapêutico , Biópsia , Comunicação Celular/efeitos dos fármacos , Comunicação Celular/imunologia , Sobrevivência Celular/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/imunologia , Ensaios de Seleção de Medicamentos Antitumorais/instrumentação , Humanos , Melanoma/sangue , Melanoma/tratamento farmacológico , Melanoma/imunologia , Cultura Primária de Células/instrumentação , Neoplasias Cutâneas/sangue , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/imunologia , Manejo de Espécimes/instrumentação , Manejo de Espécimes/métodos , Esferoides Celulares , Microambiente Tumoral/efeitos dos fármacos , Microambiente Tumoral/imunologiaRESUMO
BACKGROUND: Sun exposure is critical for vitamin D synthesis and is a major risk factor for the development of non-melanoma skin cancer (NMSC). NMSC is the most common type of cancer in Brazil and coexists with a very high prevalence of vitamin D deficiency. OBJECTIVES: The present study aimed to assess serum 25-hydroxyvitamin D (25[OH]D) concentration in patients with a recent diagnosis of NMSC. MATERIALS & METHODS: The serum 25(OH)D concentration of patients with a histological diagnosis of NMSC, made between September 2016 and September 2017, was measured by chemiluminescence up to 60 days after diagnosis and compared to healthy individuals without NMSC matched by age, sex, BMI, and skin phototype. RESULTS: Forty-one patients with NMSC and 200 healthy controls were evaluated. Most of the patients were men (56.1%) and most had basal cell carcinoma (90.2%). Patients were 67 years old on average (21-87 years) with skin Phototype 2 or 3 (80.6%). Mean serum 25(OH)D concentration in NMSC patients was significantly higher than in healthy controls (p < 0.001). Most of the patients with NMSC (65.9%) had vitamin D deficiency (25[OH]D <30 ng/mL). No association was identified between histological type, time from diagnosis, or a previous history of skin neoplasm and 25(OH)D measurements. CONCLUSION: Patients with recently diagnosed NMSC had significantly higher serum levels of 25(OH)D when compared to healthy controls. On the other hand, most of the NMSC patients were still considered to have vitamin D insufficiency.
Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Basocelular/sangue , Carcinoma de Células Escamosas/sangue , Neoplasias Cutâneas/patologia , Vitamina D/análogos & derivados , Adulto , Idoso , Idoso de 80 Anos ou mais , Análise de Variância , Brasil , Carcinoma Basocelular/patologia , Carcinoma de Células Escamosas/patologia , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica/patologia , Estadiamento de Neoplasias , Prognóstico , Valores de Referência , Fatores de Risco , Neoplasias Cutâneas/sangue , Neoplasias Cutâneas/diagnóstico , Estatísticas não Paramétricas , Vitamina D/sangueRESUMO
Skin cancer represents a serious public health problem and melanoma is considered the most significant due to its high metastasis capacity. Evasion mechanisms are the main characteristic of these tumor cells to escape of immune response. Extracellular nucleotides and nucleosides play an important role in inflammatory and immune responses. In this study, we analyzed the expression and activity of purinergic system enzymes in platelets and lymphocytes, ATP levels quantification, as well the level of pro and anti-inflammatory interleukins in the serum of 23 patients with surgical melanoma removal (CM group) and 23 control subjects (CT group). Results showed a decrease in ATP, ADP, and AMP hydrolysis and an increase in ATP levels quantification in CM group. The pro-inflammatory cytokines were elevated in CM group when compared to CT group. These results suggest an inflammatory process, even after surgical removal, due to elevated extracellular ATP levels. Besides, CM group displayed an increase in IL-10 levels and an increased in ADA activity in platelets and lymphocytes. Once adenosine and IL-10 are anti-inflammatory molecules, these results indicate a down-regulation of immune system front to malignant process. The alteration in nucleotide and nucleoside hydrolysis reinforces the purinergic systems role in this cancer. Therefore, even after surgical removal, the purinergic system can develop a chronic inflammatory micro-environment that can influence directly on relapse or metastasis.
Assuntos
Trifosfato de Adenosina/sangue , Melanoma/sangue , Neoplasias Cutâneas/sangue , Adulto , Doença Crônica , Feminino , Humanos , Inflamação/sangue , Interleucina-10/sangue , Masculino , Melanoma/patologia , Pessoa de Meia-Idade , Proteínas de Neoplasias/sangue , Neoplasias Cutâneas/patologiaAssuntos
Melanoma/sangue , Neoplasias Cutâneas/sangue , Proteína de Ligação a Vitamina D/sangue , Vitamina D/análogos & derivados , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Melanoma/patologia , Pessoa de Meia-Idade , Invasividade Neoplásica , Estudos Prospectivos , Neoplasias Cutâneas/patologia , Vitamina D/sangueRESUMO
Several studies have reported that a high neutrophil-to-lymphocyte ratio (NLR) is associated with poor clinical outcomes in several cancers, but this ratio has not been thoroughly studied in melanoma except in stage IV. This is the first study on NLR in melanoma stages I-III. This was a retrospective study of 742 melanoma patients. The NLR was classified into NLR<2 and a NLR≥2 on the basis of a receiver operating characteristic curve. Associations of NLR with clinicopathological characteristics and survival were examined. The median patient age was 57 years (range: 15-91; Q1=46, Q3=70), and the median Breslow's thickness was 3.0 mm (range: 0.5-60; Q1=1.0, Q3=7). Clinical stage at presentation was as follows: (i) stage I in 27%; (ii) stage II in 33.2%; (iii) stage III in 36.5%; and (iv) stage IV in 3.3%. NLR≥2 was associated with lymph node metastasis (36.6 vs. 18.1%) and recurrence (28.2 vs. 22.1%). The 5-year overall survival (OS) was 63% for the NLR<2 group and 53% for the NLR≥2 group. Stage-by-stage analysis showed that the 5-year OS in the NLR≥2 group for stages I, II, III, and IV were 91, 60, 28, and 0%, respectively, whereas for the NLR<2 group the 5-year OS were 98, 68, 31, and 0%, respectively. Significant differences between NLR<2 and ≥2 occurred only in stage II (P=0.014). Univariate analysis showed that factors associated with decreased OS clinical stage were Breslow's thickness, ulceration, male sex, and NLR≥2. In the multivariate analysis, all of these factors were predictors of decreased survival. The NLR appears to be an accurate prognostic marker for decreased OS in patients with melanoma, especially in clinical stage II. NLR≥2 correlated with lymph node metastasis and recurrence.
Assuntos
Linfócitos , Melanoma/sangue , Melanoma/secundário , Neutrófilos , Neoplasias Cutâneas/sangue , Neoplasias Cutâneas/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Estudos Retrospectivos , Taxa de Sobrevida , Adulto JovemRESUMO
BACKGROUND: A variety of cutaneous human papillomaviruses (HPV) are detectable in genital epithelial lesions in men and non-melanoma skin cancer patients. It remains unclear whether these viruses are associated causally with skin lesions. To date, no study has prospectively examined the association between cutaneous HPV seropositivity and development of external genital lesions (EGLs) in men. OBJECTIVES: To examine the association between seropositivity to cutaneous HPV types and the risk of subsequent development of EGLs. METHODS: A nested case-control study including 163 incident EGL cases and 352 EGL-free controls in the HPV Infection in Men (HIM) Study cohort was conducted. Cases were ascertained at each of up to 10 biannual clinical visits and verified through biopsy and pathological diagnoses. EGLs were categorized as condyloma, suggestive of condyloma, penile intraepithelial neoplasia (PeIN), and other EGLs. Archived serum specimens collected at baseline were tested for antibodies against 14 cutaneous HPV types (ß types (5, 8, 12, 14, 17, 22, 23, 24, 38, and 47), α type 27, γ type 4, µ type 1, and ν type 41) using a GST L1-based multiplex serology assay. Socio-demographic and sexual behavior data were collected through a questionnaire. Using logistic regression, adjusted odds ratios (AOR) and 95% confidence intervals (CI) were estimated. RESULTS: Overall, seropositivity to ≥1 cutaneous HPV type (any-HPV) and ≥1 ß types (any-ß) was 58.3% and 37.5% among other EGL cases, 71.6% and 46.8% among condyloma, 66.8% and 50.0% among PeIN, and 71.9% and 38.4% among controls, respectively. Type-specific seropositivity was most common for ɤ-HPV 4, µ-HPV 1, and ß-HPV 8. No statistically significant association was observed between any-HPV, any-ß, and type-specific HPV seropositivity and subsequent development of EGLs across all pathological diagnoses. CONCLUSIONS: Overall, seropositivity to cutaneous HPV was common among men; however, it appears that cutaneous HPV is not associated with the development of genital lesions in men.
Assuntos
Carcinoma in Situ/virologia , Carcinoma de Células Escamosas/virologia , Condiloma Acuminado/virologia , Genitália Masculina/virologia , Infecções por Papillomavirus/virologia , Neoplasias Penianas/virologia , Neoplasias Cutâneas/virologia , Adolescente , Adulto , Idoso , Brasil/epidemiologia , Carcinoma in Situ/sangue , Carcinoma in Situ/epidemiologia , Carcinoma de Células Escamosas/sangue , Carcinoma de Células Escamosas/epidemiologia , Estudos de Casos e Controles , Condiloma Acuminado/sangue , Condiloma Acuminado/epidemiologia , Feminino , Humanos , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/sangue , Infecções por Papillomavirus/epidemiologia , Neoplasias Penianas/sangue , Neoplasias Penianas/epidemiologia , Estudos Prospectivos , Estudos Soroepidemiológicos , Neoplasias Cutâneas/sangue , Neoplasias Cutâneas/epidemiologia , Estados Unidos/epidemiologia , Adulto JovemRESUMO
Sézary syndrome is a primary cutaneous T-cell lymphoma characterized by the triad of erythroderma, lymphadenopathy and circulating atypical cells. The emergence of new molecular targets has enabled the development of drugs such as alemtuzumab, an anti-CD52 monoclonal antibody, which has shown promising results in the treatment of this entity. We report the case of a 70-year-old male with refractory Sézary syndrome in whom treatment with alemtuzumab achieved an 80% skin lesion clearance with complete haematologic and radiologic response. The treatment was discontinued after 4 months due to adverse effects, with the patient showing a sustained response without disease progression after 13 months of follow-up.
Assuntos
Anticorpos Monoclonais Humanizados/uso terapêutico , Antineoplásicos/uso terapêutico , Síndrome de Sézary/tratamento farmacológico , Neoplasias Cutâneas/tratamento farmacológico , Idoso , Alemtuzumab , Antígenos de Diferenciação de Linfócitos T/metabolismo , Contagem de Células Sanguíneas , Humanos , Masculino , Síndrome de Sézary/sangue , Neoplasias Cutâneas/sangue , Resultado do TratamentoRESUMO
Abstract: Sézary syndrome is a primary cutaneous T-cell lymphoma characterized by the triad of erythroderma, lymphadenopathy and circulating atypical cells. The emergence of new molecular targets has enabled the development of drugs such as alemtuzumab, an anti-CD52 monoclonal antibody, which has shown promising results in the treatment of this entity. We report the case of a 70-year-old male with refractory Sézary syndrome in whom treatment with alemtuzumab achieved an 80% skin lesion clearance with complete haematologic and radiologic response. The treatment was discontinued after 4 months due to adverse effects, with the patient showing a sustained response without disease progression after 13 months of follow-up.
Assuntos
Humanos , Masculino , Idoso , Neoplasias Cutâneas/tratamento farmacológico , Síndrome de Sézary/tratamento farmacológico , Anticorpos Monoclonais Humanizados/uso terapêutico , Antineoplásicos/uso terapêutico , Neoplasias Cutâneas/sangue , Contagem de Células Sanguíneas , Antígenos de Diferenciação de Linfócitos T/metabolismo , Síndrome de Sézary/sangue , Resultado do Tratamento , AlemtuzumabRESUMO
CONTEXT AND OBJECTIVE: Kaposi's sarcoma (KS) is a common neoplastic disease in AIDS patients. The aim of this study was to evaluate the frequency of human herpesvirus 8 (HHV-8) infection in human immunodeficiency virus (HIV)-infected patients, with or without KS manifestations and correlate HHV-8 detection with KS staging. DESIGN AND SETTING: Analytic cross-sectional study conducted in a public tertiary-level university hospital in Ribeirão Preto, São Paulo, Brazil. METHODS: Antibodies against HHV-8 lytic-phase antigens were detected by means of the immunofluorescence assay. HHV-8 DNA was detected in the patient samples through a nested polymerase chain reaction (nested PCR) that amplified a region of open reading frame (ORF)-26 of HHV-8. RESULTS: Anti-HHV-8 antibodies were detected in 30% of non-KS patients and 100% of patients with KS. Furthermore, the HHV-8 DNA detection rates observed in HIV-positive patients with KS were 42.8% in serum, 95.4% in blood samples and 100% in skin biopsies; and in patients without KS, the detection rate was 4% in serum. Out of the 16 serum samples from patients with KS-AIDS who were classified as stage II, two were positive (12.5%); and out of the 33 samples from patients in stage IV, 19 (57.6%) were positive. CONCLUSION: We observed an association between HHV-8 detection and disease staging, which was higher in the serum of patients in stage IV. This suggests that detection of HHV-8 DNA in serum could be very useful for clinical assessment of patients with KS and for monitoring disease progression.
CONTEXTO E OBJETIVO: Sarcoma de Kaposi (SK) é uma doença neoplásica comum em pacientes com aids. O objetivo deste estudo foi avaliar a frequência da infecção por herpesvírus humano 8 (HHV-8) em pacientes infectados por HIV, com ou sem SK e associar a detecção do HHV-8 com o estadiamento do SK. TIPO DE ESTUDO E LOCAL: Estudo transversal analítico realizado em hospital universitário público terciário de Ribeirão Preto, São Paulo, Brasil. MÉTODOS: Anticorpos contra antígenos de fase lítica do HHV-8 foram detectados por imunofluorescência. O DNA viral de HHV-8 foi detectado em amostras de pacientes pela reação em cadeia da polimerase do tipo nested (nested PCR), que amplificou uma região do fragmento de leitura aberta (ORF)-26 do HHV-8. RESULTADOS: Anticorpos anti-HHV-8 foram detectados em 30% dos pacientes sem SK e 100% dos com SK. Além disso, a detecção de HHV-8 DNA observada em pacientes HIV-positivos com SK foi de 42,8% no soro, 95,4% em amostras de sangue e 100% em biópsias de pele, e em pacientes sem SK foi de 4% no soro. Das 16 amostras de soro de pacientes com SK-AIDS classificados como estádio II, duas foram positivas (12,5%) e, das 33 amostras de pacientes no estádio IV, 19 (57,6%) foram positivas. CONCLUSÃO: Observamos associação entre a detecção do HHV-8 e o estadiamento da doença, que foi maior no soro de pacientes no estágio IV. Isso sugere que a detecção do HHV-8 no soro poderia ser muito útil para a avaliação clínica de pacientes com SK e para o monitoramento da progressão da doença.