pI Marker Dotaz Zobrazit nápovědu
Při diagnostice nádorů štítné žlázy z folikulámích buněk jsou užívána specifická kritéria (průkaz kapsulámí a/nebo vaskuláraí invaze, morfologie buněk), která mohou být zdrojem diferenciálně diagnostických obtíží. Velká pozornost je proto nyní věnována různým markerům buněčného růstu, které by tato diagnostická úskalí eliminovaly. Cílem prezentované studie bylo stanovit proliferační MIB-1 index v souboru 155 nádorů štítné žlázy a určit, jak koreluje tento ukazatel růstu nádorových buněk s histopatologickou diagnózou, velikostí, pripadne onkocytámím vzhledem tumoru a s věkem pacienta. Onkocytámí tumory byly zastoupeny 59 folikulámími adenomy, 27 minimálně invazivními folikulámími karcinomy a 12 papilokarcinomy. Neonkocytámí nádory zahrnovaly 24 foUkidámích adenomů a 33 konvenčních papilokarcinomů. U všech tumorů byl stanoven proliferační index (PI) průkazem Ki-67 antigénu (formalin resistentního epitopu MIB-1). Ke statistickému zpracování byly použity testy ANOVA a Wilcoxonův, které prokázaly signifikantně vyšší proliferaci v karcinomech než v adenomech. Ve skupině adenomů byl navíc statisticky významný rozdíl v proliferačním indexu mezi nádory onkocytámími a neonkocytámími ve prospěch první podskupiny, což by mohlo vysvětlovat jejich častější malignizaci^ Proliferační aktivita jednotlivých typů karcinomů se naopeďi navzájem významněji nelišila. Vyšší proliferační index byl zjištěn u foliktdámích karcinomů diagnostikovaných u starších pacientů.
Well-differentiated thyroid tumors may sometimes cause diagnostic uncertainty due to difficulties in the evaluation of ceriain morphological criteria (capstdar and/or vascular invasion, cytomorphological feattires). Therefore, various diagnostic/prognostic markers are currently studied, namely the markers of tumor proliferation. The aim of our study was to evaluate the proliferative MIB-1 index in 155 thyroid tumors, and to correlate it with morphological diagnosis, size of the tumors, and the patients' age. Oncocytic tumors were represented by 59 follicular adenomas, 27 follicular carcinomas and 12 papillocarcinomas. Nononcocytic tumors comprised 24 follicular adenomas and 33 conventional papillary carcinomas. The Ki-67 antigen (formalin resistant epitope MIB-1) was detected immunohistochemically and the proliferative index (PI) of tumors was evaluated. The results were statistically analyzed using analysis of variance (ANOVA) and Wilcoxon tests (significance level p<0.05). Carcinomas showed significantly higher PI than adenomas. Moreover, PI in oncocytic adenomas was higher than in nononcocytic ones. However, proliferative activity in all types of the carcinomas was similar. The higher rates of proliferation correlated with the advanced age of the patients with follicular carcinomas (p<0.0016).
- MeSH
- adenom diagnóza ultrastruktura MeSH
- folikulární adenokarcinom diagnóza ultrastruktura MeSH
- folikulární papilární karcinom diagnóza ultrastruktura MeSH
- imunohistochemie metody MeSH
- lidé MeSH
- nádorová transformace buněk klasifikace patologie ultrastruktura MeSH
- nádorové biomarkery MeSH
- nádory štítné žlázy diagnóza patologie ultrastruktura MeSH
- Check Tag
- lidé MeSH
A new proteomic staining-free method for simultaneous identification of proteins and determination of their pI values by using low-molecular-mass pI markers is described. It is based on separation of proteins in gels by IEF in combination with mass spectrometric analysis of both peptides derived by in-gel digestion and low-molecular-mass pI markers extracted form the same piece excised from the gel. In this method, the pI markers are mixed with a protein mixture (a commercial malted barley protein extract) deposited on a gel and separated in a pH gradient. Color pI markers enable supervision of progress of focusing process. Several separated bands of the pI markers (including separated proteins) were excised and the pI markers were eluted from each gel piece by water/ethanol and identified by MALDI-TOF/TOF MS. The remaining carrier ampholytes were then washed out from gel pieces and proteins were in-gel digested with trypsin or chymotrypsin. Obtained peptides were measured by MALDI-TOF/TOF MS and proteins were identified via protein database search. This procedure allows omitting time-consuming protein staining and destaining procedures, which shortens the analysis time. For comparison, other IEF gels were stained with CBB R 250 and proteins in the gel bands were identified. Similarity of the results confirmed that our approach can give information about the correct pI values of particular proteins in complex samples at significantly shorter analysis times. This method can be very useful for identification of proteins and their post-translational modifications in prefractioned samples, where post-translational modifications (e.g., glycation) are frequent.
High-throughput mass spectrometry-based proteomic analysis requires peptide fractionation to simplify complex biological samples and increase proteome coverage. OFFGEL fractionation technology became a common method to separate peptides or proteins using isoelectric focusing in an immobilized pH gradient. However, the OFFGEL focusing process may be further optimized and controlled in terms of separation time and pI resolution. Here we evaluated OFFGEL technology to separate peptides from different samples in the presence of low-molecular-weight (LMW) color pI markers to visualize the focusing process. LMW color pI markers covering a large pH range were added to the peptide mixture before OFFGEL fractionation using a 24-wells device encompassing the pH range 3-10. We also explored the impact of LMW color pI markers on peptide fractionation labeled previously for iTRAQ. Then, fractionated peptides were separated by RP_HPLC prior to MS analysis using MALDI-TOF/TOF mass spectrometry in MS and MS/MS modes. Here we report the performance of the peptide focusing process in the presence of LMW color pI markers as on-line trackers during the OFFGEL process and the possibility to use them as pI controls for peptide focusing. This method improves the workflow for peptide fractionation in a bottom-up proteomic approach with or without iTRAQ labeling.
- MeSH
- barva MeSH
- barvicí látky chemie MeSH
- chemická frakcionace MeSH
- isoelektrická fokusace metody MeSH
- koncentrace vodíkových iontů MeSH
- lidé MeSH
- molekulová hmotnost MeSH
- peptidy analýza MeSH
- proteom analýza MeSH
- sérový albumin analýza MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- akutní poškození ledvin enzymologie moč MeSH
- biologické markery MeSH
- glutathiontransferasa moč MeSH
- ledvinové kanálky patologie MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- kongresy MeSH
BACKGROUND & AIMS: α1-Antitrypsin (AAT) is a major protease inhibitor produced by hepatocytes. The most relevant AAT mutation giving rise to AAT deficiency (AATD), the 'Pi∗Z' variant, causes harmful AAT protein accumulation in the liver, shortage of AAT in the systemic circulation, and thereby predisposes to liver and lung injury. Although intravenous AAT augmentation constitutes an established treatment of AATD-associated lung disease, its impact on the liver is unknown. METHODS: Liver-related parameters were assessed in a multinational cohort of 760 adults with severe AATD (Pi∗ZZ genotype) and available liver phenotyping, of whom 344 received augmentation therapy and 416 did not. Liver fibrosis was evaluated noninvasively via the serum test AST-to-platelet ratio index and via transient elastography-based liver stiffness measurement. Histologic parameters were compared in 15 Pi∗ZZ adults with and 35 without augmentation. RESULTS: Compared with nonaugmented subjects, augmented Pi∗ZZ individuals displayed lower serum liver enzyme levels (AST 71% vs 75% upper limit of normal, P < .001; bilirubin 49% vs 58% upper limit of normal, P = .019) and lower surrogate markers of fibrosis (AST-to-platelet ratio index 0.34 vs 0.38, P < .001; liver stiffness measurement 6.5 vs 7.2 kPa, P = .005). Among biopsied participants, augmented individuals had less pronounced liver fibrosis and less inflammatory foci but no differences in AAT accumulation were noted. CONCLUSIONS: The first evaluation of AAT augmentation on the Pi∗ZZ-related liver disease indicates liver safety of a widely used treatment for AATD-associated lung disease. Prospective studies are needed to confirm the beneficial effects and to demonstrate the potential efficacy of exogenous AAT in patients with Pi∗ZZ-associated liver disease.
- MeSH
- deficit alfa1-antitrypsinu * komplikace farmakoterapie MeSH
- dospělí MeSH
- fenotyp MeSH
- genotyp MeSH
- jaterní cirhóza etiologie MeSH
- lidé MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Poorly differentiated thyroid carcinoma (PDTC) and anaplastic thyroid carcinoma (ATC) are very rare tumors with extremely aggressive behavior. Their comprehensive genetic background is still unclear. Some of the main genetic changes of differentiated thyroid carcinomas, such as mutations in BRAF and RAS genes, as well as changes in CTNNB1, PIK3CA, TP53, AXIN1, PTEN or APC genes leading to the dedifferentiation of the tumors, are described. MATERIALS AND METHODS: DNAs from fresh frozen thyroid tissues of 3 PDTCs and 5 ATCs were extracted. The next-generation sequencing (NGS) approach was used to target 94 genes involved in cancer. The samples were prepared using a TruSight Cancer panel and sequenced with a MiSeq sequencer. Analysis of variants was performed by the MiSeq Reporter and NextGENe software and stringent criteria for prioritization of the variants were used in the Illumina VariantStudio software. RESULTS: Using NGS, we identified 26 genetic changes in 18 genes, novel variants included. CONCLUSION: NGS is a useful tool for searching for new variants and genes involved in PDTC and ATC. It seems that each of these rare tumor types has its own specific genetic background. These data could be helpful for recognizing new genetic markers and targets for future personalized therapy.
- MeSH
- anaplastický karcinom štítné žlázy genetika patologie MeSH
- buněčná diferenciace genetika MeSH
- fosfatidylinositol-3-kinasy genetika MeSH
- fosfohydroláza PTEN genetika MeSH
- lidé MeSH
- mutace MeSH
- nádorové biomarkery genetika izolace a purifikace MeSH
- nádorové proteiny genetika MeSH
- nádory štítné žlázy genetika patologie MeSH
- protoonkogenní proteiny B-raf genetika MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Somatic mutations of genes involved in NF-κB, PI3K/AKT, NOTCH, and JAK/STAT signaling pathways play an important role in the pathogenesis of Hodgkin lymphoma (HL). HL tumor cells form only about 5% of the tumor mass; however, it was shown that HL tumor-derived DNA could be detected in the bloodstream. This circulating tumor DNA (ctDNA) reflects the genetic profile of HL tumor cells and can be used for qualitative and quantitative analysis of tumor-specific somatic DNA mutations within the concept of liquid biopsy. Overall, the most frequently mutated gene in HL is STAT6; however, the exact spectrum of mutations differs between individual HL histological subtypes. Importantly, reduction of ctDNA plasma levels after initial treatment is highly correlated with prognosis. Therefore, ctDNA shows great promise as a novel tool for non-invasive tumor genome analysis for biomarker driven therapy as well as for superior minimal residual disease monitoring and treatment resistance detection. Here, we summarize the recent advancements of ctDNA analysis in HL with focus on ctDNA detection methodologies, genetic profiling of HL and its clonal evolution, and the emerging prognostic value of ctDNA.
- MeSH
- cirkulující nádorová DNA * genetika MeSH
- DNA nádorová genetika MeSH
- fosfatidylinositol-3-kinasy MeSH
- Hodgkinova nemoc * diagnóza genetika MeSH
- lidé MeSH
- mutace MeSH
- nádorové biomarkery genetika MeSH
- NF-kappa B MeSH
- protoonkogenní proteiny c-akt MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Východiska: Kombinace moderní chemoterapie s cílenou anti-EGFR léčbou vede u senzitivních pacientů s metastatickým kolorektálním karcinomem k prodloužení života a zlepšení jeho kvality. U rezistentních pacientů může však přidání monoklonálních protilátek proti EGFR vést naopak ke zhoršení parametrů přežití. Z tohoto důvodu se identifikace senzitivních a rezistentních pacientů stala klíčovou záležitostí při iniciální rozvaze před zahájením cílené léčby metastatického kolorektálního karcinomu. Četné klinické studie vedly ke zjištění, že rezistence k anti-EGFR terapii je v naprosté většině případů spojena s trvalou aktivací signálních drah distálně od EGFR. Z mnoha studovaných faktorů (mutace onkogenů KRAS, NRAS, BRAF a PIK3CA, inaktivace nádorového supresoru PTEN a TP53, amplifikace EGFR a HER2, zvýšená hladina ligandů epiregulinu a amphiregulinu, mikroRNA miR-31-3p a miR-31-5p a další) se do rutinní klinické praxe dostaly pouze KRAS a NRAS. U ostatních faktorů je třeba dalších studií k verifikaci zjištěných závěrů. Na pokračující efektivitu anti-EGFR terapie mohou ukazovat i některé klinické parametry zjištěné až po zahájení cílené léčby, jako např. časná regrese nádoru, hloubka nádorové odpovědi či míra poklesu plazmatické hladiny hořčíku. Přesnost prediktivní diagnostiky lze zvýšit rovněž kombinací vyšetřovaných biomarkerů např. pomocí metod založených na sekvenování nové generace. Je však třeba varovat před nekritickým vyšetřováním řady molekulárních markerů, které může vést k problémům s interpretací získaných výsledků, především jejich klinické relevance. Cíl: Cílem tohoto přehledu je popsat současné možnosti predikce odpovědi na anti-EGFR terapii v kontextu EGFR signální dráhy a návaznosti na běžnou klinickou praxi.
Background: The combination of modern systemic chemotherapy and anti-EGFR monoclonal antibodies improves overall survival and quality of life for patients with metastatic colorecal cancer. By contrast, the addition of anti-EGFR therapy to the treatment regime of resistant patients may lead to worse progression-free survival and overall survival. Therefore, identifying sensitive and resistant patients prior to targeted therapy of metastatic colorecal cancer is a key point during the initial decision making process. Previous research shows that primary resistance to EGFR blockade is in most cases caused by constitutive activation of signaling pathways downstream of EGFR. Of all relevant factors (mutation of KRAS, NRAS, BRAF, and PIK3CA oncogenes, inactivation of tumor suppressors PTEN and TP53, amplification of EGFR and HER2, and expression of epiregulin and amphiregulin, mikroRNA miR-31-3p, and miR-31-5p), only evaluation of KRAS and NRAS mutations has entered routine clinical practice. The role of the other markers still needs to be validated. The ongoing benefit of anti-EGFR therapy could be indicated by specific clinical parameters measured after the initiation of targeted therapy, including early tumor shrinkage, the deepness of the response, or hypomagnesemia. The accuracy of predictive diagnostic tools could be also increased by examining a combination of predictive markers using next generation sequencing methods. However, unjustified investigation of many molecular markers should be resisted as this may complicate interpretation of the results, particularly in terms of their specific clinical relevance. Aim: The aim of this review is to describe current possibilities with respect to predicting responses to EGFR blockade in the context of the EGFR pathway, and the utilization of such results in routine clinical practice. Key words: colorectal cancer – cetuximab – panitumumab – EGFR – KRAS – BRAF The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 20. 3. 2016 Accepted: 19. 4. 2016
- MeSH
- analýza přežití MeSH
- erbB receptory * antagonisté a inhibitory MeSH
- exantém MeSH
- exprese genu MeSH
- fosfatidylinositol-3-kinasy genetika MeSH
- fosfohydroláza PTEN genetika MeSH
- genetické testování * MeSH
- kolorektální nádory * genetika terapie MeSH
- kombinovaná terapie MeSH
- lidé MeSH
- metastázy nádorů genetika MeSH
- nádorové biomarkery MeSH
- nedostatek hořčíku MeSH
- prediktivní hodnota testů MeSH
- prognóza MeSH
- protokoly protinádorové kombinované chemoterapie MeSH
- protoonkogenní proteiny B-raf genetika MeSH
- ras proteiny * genetika MeSH
- retrospektivní studie MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
Purpose: Uterine sarcomas are rare and heterogeneous tumors characterized by an aggressive clinical behavior. Their high rates of recurrence and mortality point to the urgent need for novel targeted therapies and alternative treatment strategies. However, no molecular prognostic or predictive biomarkers are available so far to guide choice and modality of treatment.Experimental Design:We investigated the expression of several druggable targets (phospho-S6S240ribosomal protein, PTEN, PDGFR-α, ERBB2, and EGFR) in a large cohort of human uterine sarcoma samples (288), including leiomyosarcomas, low-grade and high-grade endometrial stromal sarcomas, undifferentiated uterine sarcomas, and adenosarcomas, together with 15 smooth muscle tumors of uncertain malignant potential (STUMP), 52 benign uterine stromal tumors, and 41 normal uterine tissues. The potential therapeutic value of the most promising target, p-S6S240, was tested in patient-derived xenograft (PDX) leiomyosarcoma models.Results:In uterine sarcomas and STUMPs, S6S240phosphorylation (reflecting mTOR pathway activation) was associated with higher grade (P= 0.001) and recurrence (P= 0.019), as shown by logistic regression. In addition, p-S6S240correlated with shorter progression-free survival (P= 0.034). Treatment with a dual PI3K/mTOR inhibitor significantly reduced tumor growth in 4 of 5 leiomyosarcoma PDX models (with tumor shrinkage in 2 models). Remarkably, the 4 responding models showed basal p-S6S240expression, whereas the nonresponding model was scored as negative, suggesting a role for p-S6S240in response prediction to PI3K/mTOR inhibition.Conclusions:Dual PI3K/mTOR inhibition represents an effective therapeutic strategy in uterine leiomyosarcoma, and p-S6S240expression is a potential predictive biomarker for response to treatment.Clin Cancer Res; 23(5); 1274-85. ©2017 AACR.
- MeSH
- cílená molekulární terapie MeSH
- fosfatidylinositol-3-kinasy genetika MeSH
- fosforylace MeSH
- inhibitory fosfoinositid-3-kinasy MeSH
- leiomyosarkom farmakoterapie genetika patologie MeSH
- lidé MeSH
- myši MeSH
- nádorové biomarkery genetika MeSH
- nádory dělohy farmakoterapie genetika patologie MeSH
- přežití bez známek nemoci MeSH
- prognóza MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- ribozomální protein S6 genetika MeSH
- signální transdukce účinky léků MeSH
- TOR serin-threoninkinasy antagonisté a inhibitory genetika MeSH
- xenogenní modely - testy protinádorové aktivity MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
