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1.
Tumour Biol ; 42(4): 1010428320916314, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32338581

RESUMO

In vitro characterization of cell-free DNA using two-dimensional cell culture models is emerging as an important step toward an improved understanding of the physical and biological characteristics of cell-free DNA in human biology. However, precise measurement of the cell-free DNA in cell culture medium is highly dependent on the efficacy of the method used for DNA purification, and is often a juncture of experimental confusion. Therefore, in this study, we compared six commercially available cell-free DNA isolation kits for the recovery of cell-free DNA from the cell culture supernatant of a human bone cancer cell line (143B), including two magnetic bead-based manual kits, one automated magnetic bead-based extraction method, and three manual spin-column kits. Based on cell-free DNA quantitation and sizing, using the Qubit dsDNA HS assay and Bioanalyzer HS DNA assay, respectively, the different methods showed significant variability concerning recovery, reproducibility, and size discrimination. These findings highlight the importance of selecting a cell-free DNA extraction method that is appropriate for the aims of a study. For example, mutational analysis of cell-free DNA may be enhanced by a method that favors a high yield or is biased toward the isolation of short cell-free DNA fragments. In contrast, quantitative analysis of cell-free DNA in a comparative setting (e.g. measuring the fluctuation of cell-free DNA levels over time) may require the selection of a cell-free DNA isolation method that forgoes a high recovery for high reproducibility and minimal size bias.


Assuntos
Ácidos Nucleicos Livres/isolamento & purificação , Meios de Cultivo Condicionados/análise , Biópsia Líquida/métodos , Biópsia Líquida/normas , Biomarcadores Tumorais , Células Cultivadas , DNA de Neoplasias , Humanos , Neoplasias/diagnóstico , Neoplasias/genética , Neoplasias/metabolismo , Reprodutibilidade dos Testes
2.
Medicine (Baltimore) ; 99(8): e19097, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32080083

RESUMO

Liquid biopsy is an emerging technique for noninvasive detection of various cancers. Majority of liquid biopsy tests still, however, use solitary type of biomarkers with unsatisfactory sensitivity and specificity. To this end, a combined approach of circulating tumor cells (CTCs) and salivary mRNA biomarkers was evaluated for discriminating non-small-cell lung cancer (NSCLC) from healthy controls.Our study included a discovery phase to find multiple biomarkers, and an independent validation phase to confirm the applicability of the selected biomarkers. In the discovery phase, CTC level in blood and 5 mRNA biomarkers in saliva (i.e., CCNI, Epidermal growth factor receptor [EGFR], FGF19, FRS2, and GREB1) were measured for 140 NSCLC patients and 140 healthy controls, followed by developing a predictive model. Next, this panel of biomarkers was applied to another patient cohort consisted of 60 patients with NSCLC and 60 healthy controls in the validation phase.We found that our novel biomarker panel could differentiate patients with NSCLC from healthy controls with high sensitivity (92.1%) and high specificity (92.9%) in the discovery phase. In the validation phase, we achieved sensitivity of 88.3% and specificity of 90.0%.To our best knowledge, it is the first time that a combined use of CTC and salivary mRNA biomarkers were applied for noninvasive detection of NSCLC.


Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Células Neoplásicas Circulantes/metabolismo , RNA Mensageiro/metabolismo , Saliva/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Adulto , Idoso , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Ciclina I/metabolismo , Receptores ErbB/metabolismo , Feminino , Fatores de Crescimento de Fibroblastos/metabolismo , Humanos , Biópsia Líquida/métodos , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Masculino , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Proteínas de Neoplasias/metabolismo , Células Neoplásicas Circulantes/patologia , Valor Preditivo dos Testes , Sensibilidade e Especificidade
3.
Crit Rev Oncol Hematol ; 146: 102879, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32005411

RESUMO

Cell-free circulating tumor DNA (ct-DNA) reflecting the whole tumor spatial and temporal heterogeneity currently represents the most promising candidate for liquid biopsy strategy in glioma. Unlike other solid tumors, it is now widely accepted that the best source of ct-DNA for glioma patients is the cerebrospinal fluid, since blood levels are usually low and detectable only in few cases. A cerebrospinal fluid ct-DNA liquid biopsy approach may virtually support all the stages of glioma management, from facilitating molecular diagnosis when surgery is not feasible, to monitoring tumor response, identifying early recurrence, tracking longitudinal genomic evolution, providing a new molecular characterization at recurrence and allowing patient selection for targeted therapies. This review traces the history of ct-DNA liquid biopsy in the field of diffuse malignant gliomas, describes its current status and analyzes what are the future perspectives and pitfalls of this potentially revolutionary molecular tool.


Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Encefálicas/líquido cefalorraquidiano , DNA Tumoral Circulante/líquido cefalorraquidiano , DNA de Neoplasias/metabolismo , Glioma/líquido cefalorraquidiano , Biópsia Líquida/métodos , Células Neoplásicas Circulantes/metabolismo , Biomarcadores Tumorais/genética , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patologia , DNA de Neoplasias/genética , Genes Neoplásicos/genética , Glioblastoma/líquido cefalorraquidiano , Glioblastoma/genética , Glioblastoma/patologia , Glioma/genética , Glioma/patologia , Humanos , Mutação , Gradação de Tumores , Recidiva Local de Neoplasia/diagnóstico , Recidiva Local de Neoplasia/genética , Recidiva Local de Neoplasia/metabolismo , Células Neoplásicas Circulantes/patologia
4.
Medicine (Baltimore) ; 99(3): e18581, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32011436

RESUMO

BACKGROUND: Liquid biopsy is a novel method for cancer diagnosis, which has been applied in lung and breast cancers, demonstrating high diagnostic value. However, clinical value of it in pancreatic cancer (PC) remains to be verified. The aim of this meta-analysis was to evaluate overall diagnostic value of various liquid biopsy methods (circulating tumor DNA, circulating tumor cells and exosomes) in detecting PC. METHODS: We comprehensively searched relevant studies in PubMed, Medline, Embase, and Web of Science without time limitation according to PRISMA. Data necessary for reconstructing a 2 × 2 table was calculated from the original articles. The methodological quality of included studies was evaluated by QUADAS-2. Statistical analysis including was performed by the software Meta-Disc version 1.4, and STATA 14.2. RESULTS: A total of 19 studies including 1872 individuals were included in this meta-analysis. In which, 7 were studies about ctDNA, 7 were on CTCs and 6 were about exosomes (Sefrioui D, studied diagnostic accuracy of both ctDNA and CTCs, with no common patients in these 2 groups). The pooled sensitivity estimates for ctDNA, CTCs and exosomes in detecting PC with their 95% confidential intervals (95% CI) were 0.64 (95%CI 0.58-0.70), 0.74 (95%CI 0.68-0.79) and 0.93 (95%CI 0.90-0.95), respectively. The pooled specificity estimates were 0.92(95%CI 0.88-0.95), 0.83 (95%CI 0.78-0.88) and 0.92 (95%CI 0.88-0.95), respectively. The area under curve (AUC) of the sROC for ctDNA, CTCs and exosomes in detecting PC were 0.9478, 0.8166, and 0.9819, respectively. The overall sensitivity, specificity and AUC of the sROC curve for overall liquid biopsy in detecting PC were 0.80 (95%CI 0.77-0.82), 0.89 (95%CI 0.87-0.91) and 0.9478, respectively. CONCLUSION: This meta-analysis confirmed that liquid biopsy had high diagnostic value in detecting PC. In ctDNA, CTCs and exosomes these 3 subgroups, exosomes showed highest sensitivity and specificity.


Assuntos
Biópsia Líquida/métodos , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/patologia , Biomarcadores Tumorais , DNA de Neoplasias , Exossomos , Humanos , Biópsia Líquida/normas , Neoplasias Pancreáticas/genética , Sensibilidade e Especificidade
5.
Crit Rev Oncol Hematol ; 146: 102863, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31935617

RESUMO

Lung cancer is the most frequent cancer for males and third most frequent cancer for females. Targeted therapy drugs based on molecular alterations, such as angiogenesis inhibitors, epidermal growth factor receptor (EGFR) inhibitors, and anaplastic lymphoma kinase (ALK) inhibitors are important part of treatment of NSCLC. However, the quality of the available tumor biopsy and/or cytology material is sometimes not adequate to perform the necessary molecular testing, which has prompted the search for alternatives. This review examines the use of tumor-educated platelet (TEP) as a liquid biopsy in lung cancer patients. The development of sensitive and accurate techniques have made it possible to detect the specific genetic alterations for which targeted therapies are already available. Liquid biopsy offers opportunities to detect resistance mechanisms at an early stage. To conclude, tumor-educated platelet has the potential to be used as liquid biopsy for a variety of clinical and investigational applications.


Assuntos
Biomarcadores Tumorais/sangue , Carcinoma Pulmonar de Células não Pequenas/patologia , Ácidos Nucleicos Livres/sangue , DNA de Neoplasias/sangue , DNA de Neoplasias/genética , Biópsia Líquida/métodos , Neoplasias Pulmonares/patologia , Biomarcadores Tumorais/genética , Plaquetas/patologia , Carcinoma Pulmonar de Células não Pequenas/sangue , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/terapia , Ácidos Nucleicos Livres/genética , Receptores ErbB/genética , Feminino , Humanos , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/terapia , Masculino , Mutação , Medicina de Precisão , Inibidores de Proteínas Quinases/uso terapêutico
6.
Cancer Invest ; 38(2): 85-93, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31939681

RESUMO

The identification and quantification of actionable mutations are critical for guiding targeted therapy and monitoring drug response in colorectal cancer. Liquid biopsy (LB) based on plasma cell-free DNA analysis has emerged as a noninvasive approach with many clinical advantages over conventional tissue sampling. Here, we developed a LB protocol using ultra-deep massive parallel sequencing and validated its clinical performance for detection and quantification of actionable mutations in three major driver genes (KRAS, NRAS and BRAF). The assay showed a 92% concordance for mutation detection between plasma and paired tissues and great reliability in quantification of variant allele frequency.


Assuntos
DNA Tumoral Circulante/genética , Neoplasias Colorretais/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Biópsia Líquida/métodos , Neoplasias Colorretais/sangue , GTP Fosfo-Hidrolases/genética , Humanos , Proteínas de Membrana/genética , Mutação , Proteínas Proto-Oncogênicas B-raf/genética , Proteínas Proto-Oncogênicas p21(ras)/genética , Reprodutibilidade dos Testes
7.
J Cancer Res Clin Oncol ; 146(1): 205-219, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31620896

RESUMO

BACKGROUND: Third line innovative systemic treatments and loco-regional chemotherapy by hypoxic pelvic perfusion (HPP) have both been proposed for the treatment of unresectable not responsive recurrent rectal cancer (URRC). In the present study, we have compared the safety and efficacy of HPP/target therapy, using drug regimens selected by liquid biopsy precision oncotherapy, to third-line systemic therapy based on tissue specimens precision oncotherapy. METHODS: HPP/target therapy regimens were selected based on precision oncotherapy, including assays for chemosensitivity and viability, and qRT-PCR for tumor-related gene expression. In the control group, systemic third-line and further lines of therapy were defined according to clinical and biological parameters. RESULTS: From 2007 to 2019, 62 URRC patients were enrolled, comprised of 43 patients in the HPP/target-therapy group and 19 patients in the systemic therapy control group. No HPP related complications were reported and the most common adverse events were skin and bone marrow toxicity. In the HPP/target-therapy group, the ORR was 41.8% whereas in the systemic therapy control group was 15.8%. DCR of the HPP/target-therapy group was significantly improved over the systemic therapy group (P = 0.001), associated with a PFS of 8 vs 4 months (P = 0.009), and OS of 20 vs 8 months (P = 0.046). CONCLUSIONS: The present data indicate that in URCC patients, the integration of HPP/target-therapy and precision oncotherapy based upon liquid biopsy is as effective and efficacious as third-line treatment in local disease control and, therefore, deserves to be further assessed and compared to conventional systemic treatments in future prospective randomized trials.


Assuntos
Biópsia Líquida/métodos , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/terapia , Neoplasias Retais/patologia , Neoplasias Retais/terapia , Idoso , Quimiorradioterapia Adjuvante , Estudos de Coortes , Progressão da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Terapia Neoadjuvante , Medicina de Precisão/métodos , Estudos Retrospectivos
8.
Hematol Oncol ; 38(1): 34-37, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31872890

RESUMO

This report summarizes a closed workshop cosponsored by the American Association for Cancer Research, the European School of Oncology, and the 15th-International Conference on Malignant Lymphoma to discuss critical open questions on liquid biopsy in lymphoid malignancies, develops a roadmap for their analytical and clinical validation, and prioritizes research areas.


Assuntos
Biomarcadores Tumorais/genética , DNA Tumoral Circulante/sangue , Biópsia Líquida/métodos , Linfoma/sangue , DNA Tumoral Circulante/genética , Congressos como Assunto , Humanos , Linfoma/diagnóstico , Linfoma/genética , Linfoma/terapia , Manejo de Espécimes
9.
Recent Results Cancer Res ; 215: 181-211, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31605230

RESUMO

ctDNA provided by liquid biopsy offers a promising alternative to tumor biopsy as it gives a non-invasive and «real-time¼ access to the cancer genome and reflects tumor intra and extra heterogeneity. ctDNA has shown growing clinical interest for cancer diagnosis, prognosis, theragnostics, therapeutic monitoring, and clonal evolution tracking. A major technical limit for ctDNA analysis from body fluids is the extremely low proportion of ctDNA compared to non-malignant cell-free DNA, underscoring the need for highly sensitive and specific detection techniques. The control of pre-analytical procedures appears essential for optimal ctDNA analysis and need to be standardized for clinical research applications. This chapter provides insights into major current technologies for ctDNA detection. Overall, PCR-based techniques are able to detect limited molecular alterations and have a high sensitivity suitable for monitoring purposes while NGS-based approaches are broad range molecular screening assays more specifically indicated for treatment selection. We briefly reviewed new technical innovations that are now available for ctDNA detection.


Assuntos
DNA Tumoral Circulante/análise , DNA Tumoral Circulante/isolamento & purificação , Biópsia Líquida/métodos , Neoplasias/diagnóstico , Neoplasias/genética , DNA Tumoral Circulante/genética , Humanos , Neoplasias/sangue , Neoplasias/terapia , Reação em Cadeia da Polimerase , Prognóstico
10.
Recent Results Cancer Res ; 215: 319-344, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31605237

RESUMO

Extracellular micro- and nanoscale membrane vesicles produced by different cells progressively attract the attention of the scientific community. They function as mediators of intercellular communication and transport genetic material and signaling molecules between the cells. In the context of keeping homeostasis, the extracellular vesicles contribute to the regulation of various systemic and local processes. Vesicles released by the tumor and activated stromal cells exhibit multiple functions including support of tumor growth, preparation of the pre-metastatic niches, and immune suppression. Considerable progress has been made regarding the criteria of classification of the vesicles according to their origin, content, and function: Exosomes, microvesicles, also referred to as microparticles or ectosomes, and large oncosomes were defined as actively released vesicles. Additionally, apoptotic bodies represented by a highly heterogeneous population of particles produced during apoptosis, the programmed cell death, should be considered. Because the majority of isolation techniques do not allow the separation of different types of vesicles, a joined term "extracellular vesicles" (EVs) was recommended by the ISEV community for the definition of vesicles isolated from either the cell culture supernatants or the body fluids. Because EV content reflects the content of the cell of origin, multiple studies on EVs from body fluids in the context of cancer diagnosis, prediction, and prognosis were performed, actively supporting their high potential as a biomarker source. Here, we review the leading achievements in EV analysis from body fluids, defined as EV-based liquid biopsy, and provide an overview of the main EV constituents: EV surface proteins, intravesicular soluble proteins, EV RNA including mRNA and miRNA, and EV DNA as potential biomarkers. Furthermore, we discuss recent developments in technology for quantitative EV analysis in the clinical setting and future perspectives toward miniaturized high-precision liquid biopsy approaches.


Assuntos
Vesículas Extracelulares , Biópsia Líquida/métodos , Biópsia Líquida/tendências , Neoplasias/diagnóstico , Neoplasias/patologia , Apoptose , Micropartículas Derivadas de Células , Exossomos , Humanos
11.
Cancer Metastasis Rev ; 38(4): 553-571, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31836951

RESUMO

Pediatric solid tumors have long been known to shed tumor cells, DNA, RNA, and proteins into the blood. Recent technological advances have allowed for improved capture and analysis of these typically scant circulating materials. Efforts are ongoing to develop "liquid biopsy" assays as minimally invasive tools to address diagnostic, prognostic, and disease monitoring needs in childhood cancer care. Applying these highly sensitive technologies to serial liquid biopsies is expected to advance understanding of tumor biology, heterogeneity, and evolution over the course of therapy, thus opening new avenues for personalized therapy. In this review, we outline the latest technologies available for liquid biopsies and describe the methods, pitfalls, and benefits of the assays that are being developed for children with extracranial solid tumors. We discuss what has been learned in several of the most common pediatric solid tumors including neuroblastoma, sarcoma, Wilms tumor, and hepatoblastoma and highlight promising future directions for the field.


Assuntos
Biópsia Líquida/métodos , Neoplasias/sangue , Pediatria/métodos , Criança , Humanos , Neoplasias/patologia , Ensaios Clínicos Controlados Aleatórios como Assunto
12.
Nat Commun ; 10(1): 3941, 2019 09 02.
Artigo em Inglês | MEDLINE | ID: mdl-31477699

RESUMO

Liquid biopsies have the utility for detecting minimal residual disease in several cancer types. Here, we investigate if liquid biopsy tracking on-treatment informs on tumour phenotypes by longitudinally quantifying circulating Epstein-barr virus (EBV) DNA copy number in 673 nasopharyngeal carcinoma patients undergoing radical induction chemotherapy (IC) and chemo-radiotherapy (CRT). We observe significant inter-patient heterogeneity in viral copy number clearance that is classifiable into eight distinct patterns based on clearance kinetics and bounce occurrence, including a substantial proportion of complete responders (≈30%) to only one IC cycle. Using a supervised statistical clustering of disease relapse risks, we further bin these eight subgroups into four prognostic phenotypes (early responders, intermediate responders, late responders, and treatment resistant) that are correlated with efficacy of chemotherapy intensity. Taken together, we show that real-time monitoring of liquid biopsy response adds prognostic information, and has the potential utility for risk-adapted treatment de-intensification/intensification in nasopharyngeal carcinoma.


Assuntos
Infecções por Vírus Epstein-Barr/terapia , Biópsia Líquida/métodos , Carcinoma Nasofaríngeo/terapia , Neoplasias Nasofaríngeas/terapia , Adulto , Quimiorradioterapia , DNA Viral/sangue , DNA Viral/genética , Infecções por Vírus Epstein-Barr/complicações , Infecções por Vírus Epstein-Barr/virologia , Feminino , Herpesvirus Humano 4/efeitos dos fármacos , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/efeitos da radiação , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo/complicações , Fenótipo , Prognóstico
13.
Cancer Treat Rev ; 79: 101893, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31499407

RESUMO

BACKGROUND: The management of locally advanced rectal cancer (RC) is an evolving clinical field where the multidisciplinary approach can reach its best, and liquid biopsy for obtaining tumor-derived component such as circulating tumor DNA (ctDNA) might provide complementary informations. METHODS: A systematic review of studies available in literature of liquid biopsy in non-metastatic RC has been performed according to PRISMA criteria to assess the role of ctDNA as a diagnostic, predictive and prognostic biomarker in this setting. RESULTS: Twenty-five publications have been retrieved, of which 8 full-text articles, 7 abstracts and 10 clinical trials. Results have been categorized into three groups: diagnostic, predictive and prognostic. Few but promising data are available about the use of liquid biopsy for early diagnosis of RC, with the main limitation of sensitivity due to low concentrations of ctDNA in this setting. In terms of prediction of response to chemoradiation, still inconclusive data are available about the utility of a pre-treatment liquid biopsy, whereas some studies report a positive correlation with a dynamic (pre/post-treatment) monitoring. The presence of minimal residual disease by ctDNA was consistently associated with worse prognosis across studies. CONCLUSIONS: The use of liquid biopsy for monitoring response to chemoradiation and assess the risk of disease recurrence are the most advanced potential applications for liquid biopsy in RC, with implications also in the context of non-operative management strategies.


Assuntos
Biomarcadores Tumorais , Biópsia Líquida , Neoplasias Retais/diagnóstico , DNA Tumoral Circulante , DNA de Neoplasias , Humanos , Biópsia Líquida/métodos , Metástase Neoplásica , Estadiamento de Neoplasias , Células Neoplásicas Circulantes , Prognóstico , Neoplasias Retais/etiologia , Neoplasias Retais/mortalidade , Neoplasias Retais/terapia , Recidiva , Resultado do Tratamento
14.
Int J Mol Sci ; 20(19)2019 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-31554202

RESUMO

Different collection methods may influence the ability to detect and quantify biomarker levels in saliva, particularly in the expression of DNA/RNA methylation regulators of several inflammations and tissue turnover markers. This pilot study recruited five participants and unstimulated saliva were collected by either spitting or drooling, and the relative preference for each method was evaluated using a visual analogue scale. Subsequently, total RNA, gDNA and proteins were isolated using the Trizol method. Thereafter, a systematic evaluation was carried out on the potential effects of different saliva collection methods on periodontium-associated genes, DNA/RNA epigenetic factors and periodontium-related DNA methylation levels. The quantity and quality of DNA and RNA were comparable from different collection methods. Periodontium-related genes, DNA/RNA methylation epigenetic factors and periodontium-associated DNA methylation could be detected in the saliva sample, with a similar expression for both methods. The methylation of tumour necrosis factor-alpha gene promoter from drooling method showed a significant positive correlation (TNF α, r = 0.9) with clinical parameter (bleeding on probing-BOP). In conclusion, the method of saliva collection has a minimal impact on detecting periodontium-related genetic and epigenetic regulators in saliva. The pilot data shows that TNF α methylation may be correlated with clinical parameters.


Assuntos
Biomarcadores , Epigênese Genética , Epigenômica , Testes Genéticos , Periodonto/metabolismo , Saliva/metabolismo , Adulto , DNA , Metilação de DNA , Epigenômica/métodos , Feminino , Perfilação da Expressão Gênica , Testes Genéticos/métodos , Humanos , Biópsia Líquida/métodos , Masculino , Metilação , Projetos Piloto , Proteínas , RNA , Adulto Jovem
15.
Asian Pac J Cancer Prev ; 20(9): 2611-2617, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31554354

RESUMO

Objective: Immunocytochemistry (ICC) of serous effusion is an important tool for the diagnosis of benign and malignant cells. Our aim was to develop a modified liquid-based cytological technique for ICC (i.e., a modified LBC). Methods: Serous effusions of 110 cases were collected for cytological examination: 50 were negative for malignancy albeit benign mesothelium was found, and 60 were confirmed metastatic adenocarcinoma according to the modified LBC preparation. The latter were stained for EMA, Ber-EP4, Calretinin, and p63 then interpreted by both a cytotechnologist and a pathologist. A comparative analysis of the diagnostic results was conducted. Results: The results of the metastatic adenocarcinoma were 100% (60/60) positive for EMA and 91.7% (55/60) positive for Ber-Ep4 but negative for calretinin and p63. Cases negative for malignancy were 100% (50/50) positive for calretinin but negative for carcinoma markers. The difference between 'positive for metastatic adenocarcinoma' and 'negative for malignancy' in ICC was statistically significant (p < 0.001). Conclusion: The current study demonstrated that a panel marker, comprising EMA, Ber-EP4, and calretinin can be used for differentiating between cases of metastatic adenocarcinoma and benign mesothelium. The serous effusion specimen collected by the modified LBC technique is an effective preparation method for ICC.


Assuntos
Adenocarcinoma/secundário , Citodiagnóstico/métodos , Imuno-Histoquímica/métodos , Biópsia Líquida/métodos , Mesotelioma/patologia , Derrame Pleural Maligno/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais , Feminino , Seguimentos , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Prognóstico
16.
World J Gastroenterol ; 25(29): 3985-3995, 2019 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-31413532

RESUMO

BACKGROUND: Hepatitis B is a major public health problem in China. Accurate liver injury assessment is essential for clinical evidence-based treatment. Liver biopsy is considered the gold standard method to stage liver disease, but it is not widely used in resource-limited settings. Therefore, non-invasive liquid biopsy tests are needed. AIM: To assess liver injury in hepatitis B patients using quantified cell free DNA combined with other serum biomarker as a liquid biopsy-based method. METHODS: A cohort of 663 subjects including 313 hepatitis B patients and 350 healthy controls were enrolled. Ultrasound-guided liver biopsies followed by histopathological assessments were performed for the 263 chronic hepatitis B patients to determine the degree of liver injury. Cell-free DNA was quantified using a novel duplex real-time polymerase chain reaction assay. RESULTS: Compared with healthy controls, patients with hepatitis B virus (HBV) infection had significantly higher plasma DNA, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, and HBV DNA levels (P < 0.01). Serum ALT, AST, bilirubin, and plasma DNA levels of patients with marked-severe inflammation were significantly higher than those with mild-moderate inflammation (P < 0.01). There was a statistically significant correlation between hepatocyte inflammation severity and serum bilirubin (R 2 = 0.673, P < 0.01) or plasma DNA (R 2 = 0.597, P < 0.01) levels. The areas under the curves of serum ALT, bilirubin, plasma DNA, and their combination to distinguish between patients with mild-moderate and marked-severe inflammation were 0.8059, 0.7910, 0.7921, and 0.9564, respectively. CONCLUSION: The combination of plasma DNA, serum ALT, and bilirubin could be a candidate liquid biopsy for non-invasive assessment of liver injury in hepatitis B patients.


Assuntos
Hepatite B Crônica/diagnóstico , Testes de Função Hepática/métodos , Fígado/patologia , Adolescente , Adulto , Idoso , Alanina Transaminase/sangue , Bilirrubina/sangue , Biomarcadores/sangue , Ácidos Nucleicos Livres/sangue , China , Estudos de Coortes , Estudos de Viabilidade , Feminino , Voluntários Saudáveis , Hepatite B Crônica/sangue , Hepatite B Crônica/patologia , Humanos , Biópsia Líquida/métodos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Índice de Gravidade de Doença , Adulto Jovem
18.
Nat Commun ; 10(1): 3856, 2019 08 26.
Artigo em Inglês | MEDLINE | ID: mdl-31451693

RESUMO

Accurate prediction of chemo- or targeted therapy responses for patients with similar driver oncogenes through a simple and least-invasive assay represents an unmet need in the clinical diagnosis of non-small cell lung cancer. Using a single-cell on-chip metabolic cytometry and fluorescent metabolic probes, we show metabolic phenotyping on the rare disseminated tumor cells in pleural effusions across a panel of 32 lung adenocarcinoma patients. Our results reveal extensive metabolic heterogeneity of tumor cells that differentially engage in glycolysis and mitochondrial oxidation. The cell number ratio of the two metabolic phenotypes is found to be predictive for patient therapy response, physiological performance, and survival. Transcriptome analysis reveals that the glycolytic phenotype is associated with mesenchymal-like cell state with elevated expression of the resistant-leading receptor tyrosine kinase AXL and immune checkpoint ligands. Drug targeting AXL induces a significant cell killing in the glycolytic cells without affecting the cells with active mitochondrial oxidation.


Assuntos
Adenocarcinoma de Pulmão/diagnóstico , Neoplasias Pulmonares/diagnóstico , Metabolômica/métodos , Derrame Pleural Maligno/patologia , Análise de Célula Única/métodos , Adenocarcinoma de Pulmão/mortalidade , Adenocarcinoma de Pulmão/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígeno B7-H1/metabolismo , Biomarcadores Tumorais/metabolismo , Contagem de Células , Feminino , Perfilação da Expressão Gênica/métodos , Humanos , Estimativa de Kaplan-Meier , Biópsia Líquida/métodos , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/patologia , Masculino , Análise em Microsséries/métodos , Pessoa de Meia-Idade , Prognóstico , Proteína 2 Ligante de Morte Celular Programada 1/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Receptores Proteína Tirosina Quinases/metabolismo
19.
Anal Bioanal Chem ; 411(23): 6039-6047, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31304564

RESUMO

Circulating tumor DNA (ctDNA) is a tumor-derived fragmented DNA in the bloodstream that is not associated with cells. It has been greatly focused in the recent decade because of its potential clinical utility for liquid biopsies. Development of ctDNA analytical techniques with high sensitivity and cost-efficiency will undoubtedly promote the clinical spread of ctDNA testing. In this paper, we propose a novel flow cytometry-based ctDNA sensing strategy which combines enzyme-free amplification and magnetic separation. The target DNA is capable of triggering a hybridization chain reaction, producing a fluorescent long linear assembly of DNA, which can be further captured by magnetic beads to present fluorescent signals using flow cytometry. In comparison with some conventional methods, our strategy has the advantages of easy operation and cost-efficiency, and thereby shows a promising application in clinical diagnosis. Graphical abstract.


Assuntos
DNA Tumoral Circulante/sangue , Citometria de Fluxo/métodos , Imãs/química , Técnicas de Amplificação de Ácido Nucleico/métodos , DNA Tumoral Circulante/análise , Humanos , Limite de Detecção , Biópsia Líquida/métodos , Hibridização de Ácido Nucleico/métodos
20.
Vet Clin North Am Small Anim Pract ; 49(5): 781-791, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31280902

RESUMO

Molecular diagnostics have revolutionized human oncology to allow early detection, targeted therapy, monitoring throughout treatment, and evidence of recurrence. By identifying genetic signatures associated with cancers, liquid biopsy techniques have been developed to diagnose and monitor cancer in noninvasive or minimally invasive ways. These techniques offer new opportunities for improving cancer screening, diagnosis, and monitoring the impact of therapy on the patients over time. Liquid biopsy also drives drug development programs. Similar diagnostics hold promise for comparable results in the veterinary field. Several noninvasive/minimally invasive techniques have been described in veterinary medicine that could be referred to as liquid biopsy.


Assuntos
Doenças do Cão/diagnóstico , Biópsia Líquida/veterinária , Neoplasias/veterinária , Animais , Carcinoma de Células de Transição/diagnóstico , Carcinoma de Células de Transição/genética , Carcinoma de Células de Transição/veterinária , Doenças do Cão/tratamento farmacológico , Doenças do Cão/genética , Cães , Detecção Precoce de Câncer/métodos , Detecção Precoce de Câncer/veterinária , Feminino , Humanos , Leucemia/diagnóstico , Leucemia/veterinária , Biópsia Líquida/métodos , Linfoma/diagnóstico , Linfoma/tratamento farmacológico , Linfoma/veterinária , Masculino , Terapia de Alvo Molecular/veterinária , Mutação , Neoplasias/diagnóstico , Neoplasias/tratamento farmacológico , Neoplasias/genética , Neoplasias Uretrais/diagnóstico , Neoplasias Uretrais/genética , Neoplasias Uretrais/veterinária , Neoplasias da Bexiga Urinária/veterinária
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