BACKGROUND: This is a multicentre, European, prospective trial evaluating the diagnostic accuracy of One Step Nucleic Acid Amplification (OSNA) compared to sentinel lymph nodes histopathological ultrastaging in endometrial cancer patients. METHODS: Centres with expertise in sentinel lymph node mapping in endometrial cancer patients in Europe will be invited to participate in the study. Participating units will be trained on the correct usage of the OSNA RD-210 analyser and nucleic acid amplification reagent kit LYNOAMP CK19 E for rapid detection of metastatic nodal involvement, based on the cytokeratin 19 (CK19) mRNA detection. Endometrial cancer patients ≥ 18 years listed for surgical treatment with sentinel lymph node mapping, with no history of other types of cancer and who provide a valid written consent will be considered potentially eligible for the study. However, they will only be enrolled if a successful sentinel lymph node mapping is retrieved. Each node will be processed according to the study protocol and assessed by both OSNA and ultrastaging. DISCUSSION: The accuracy of OSNA (index test) will be assessed against sentinel lymph node histopathological ultrastaging (reference test). This European study has the potential to be the largest study on the use of OSNA in endometrial cancer to date. OSNA could represent a modern diagnostic alternative to sentinel lymph node ultrastaging with the added benefits of standardisation and fast results. TRIAL REGISTRATION: The study was registered in the German Clinical Trial Register - Nr. DRKS00021520, registration date 25th of May 2020, URL of the trial registry record: https://drks.de/search/en/trial/DRKS00021520 .
- Klíčová slova
- Endometrial cancer, OSNA, Sentinel lymph nodes, Ultrastaging,
- MeSH
- biopsie sentinelové lymfatické uzliny metody MeSH
- keratin-19 genetika MeSH
- lidé MeSH
- lymfatické metastázy * diagnóza patologie MeSH
- lymfatické uzliny patologie MeSH
- multicentrické studie jako téma MeSH
- nádory endometria * patologie genetika diagnóza MeSH
- prospektivní studie MeSH
- sentinelová uzlina * patologie MeSH
- staging nádorů MeSH
- techniky amplifikace nukleových kyselin * metody MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- protokol klinické studie MeSH
- Geografické názvy
- Evropa MeSH
- Názvy látek
- keratin-19 MeSH
The aim of the study was to compare the prognostic significance of lymph node status of patients with lung cancer analyzed by three different methods: hematoxylin and eosin (H&E), immunohistochemistry of cytokeratin 19 (IHC CK19), and One-Step Nucleic Acid Amplification (OSNA). The clinical relevance of the results was evaluated based on relation to prognosis; the disease-free interval (DFI) and overall survival (OS) were analyzed. During radical surgical treatment, a total of 1426 lymph nodes were obtained from 100 patients, creating 472 groups of nodes (4-5 groups per patient) and examined by H&E, IHC CK19 and OSNA. The median follow-up was 44 months. Concordant results on the lymph node status of the H&E, IHC CK19 and OSNA examinations were reported in 78% of patients. We recorded shorter OS in patients with positive results provided by both OSNA and H&E. The study demonstrated a higher percentage of detected micrometastases in lymph nodes by the OSNA method. However, the higher sensitivity of the OSNA, with the cut-off value 250 copies of mRNA of CK19/µL, resulted in a lower association of OSNA positivity with progress of the disease compared to H&E. Increasing the cut-off to 615 copies resulted in an increase in concordance between the OSNA and H&E, which means that the higher cut-off is more relevant in the case of lung tumors.
- Klíčová slova
- E, H&, IHC CK19, OSNA, lung cancer, lymph nodes,
- MeSH
- imunohistochemie metody MeSH
- keratin-19 genetika MeSH
- lidé MeSH
- lymfatické metastázy * MeSH
- lymfatické uzliny patologie MeSH
- metastázy nádorů MeSH
- nádorové biomarkery genetika MeSH
- nádory plic diagnóza patologie chirurgie MeSH
- následné studie MeSH
- přežití bez známek nemoci MeSH
- prognóza MeSH
- prospektivní studie MeSH
- staging nádorů MeSH
- techniky amplifikace nukleových kyselin metody MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- keratin-19 MeSH
- nádorové biomarkery MeSH
BACKGROUND AND OBJECTIVES: Utilisation of the one-step nucleic acid amplification (OSNA) molecular biology method for the detection of the metastatic involvement of sentinel lymph nodes (SLNs) in endometrial cancer (EC) patients. A comparison with histopathological ultrastaging and a description of the clinical consequences. METHODS: Surgically treated EC patients underwent detection of SLNs. Nodes greater than 5 mm were cut into sections 2-mm thick parallel to the short axis of the node. Odd sections were examined according to the OSNA method, while even ones according to an appropriate ultrastaging protocol. Nodes less than or equal to 5 mm were cut into halves along the longitudinal axis with one half examined according to the OSNA method and the other half by ultrastaging. RESULTS: Fifty-eight patients were included and 135 SLNs were acquired. Both ultrastaging and OSNA agreed on 116 results. According to the OSNA method, 20.69% more patients were classified into International Federation of Gynecology and Obstetrics (FIGO) stage III. When comparing the results of the OSNA method to the conclusions of ultrastaging as a reference method, sensitivity of 90.9%, specificity of 85.5% and concordance of 85.9% were attained. CONCLUSIONS: The results of the OSNA method showed a higher frequency of detection of micrometastases and included 20.69% more patients into FIGO stage III.
- Klíčová slova
- cytokeratin 19, micrometastasis, one-step nucleic acid amplification assay,
- MeSH
- adenokarcinom z jasných buněk genetika sekundární chirurgie MeSH
- biopsie sentinelové lymfatické uzliny MeSH
- dospělí MeSH
- keratin-19 genetika MeSH
- lidé středního věku MeSH
- lidé MeSH
- lymfatické metastázy MeSH
- mikrometastázy MeSH
- míra přežití MeSH
- nádorové biomarkery genetika MeSH
- nádory endometria genetika patologie chirurgie MeSH
- následné studie MeSH
- nukleové kyseliny analýza genetika MeSH
- peroperační doba MeSH
- prognóza MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- sentinelová uzlina patologie chirurgie MeSH
- serózní cystadenokarcinom genetika sekundární chirurgie MeSH
- techniky amplifikace nukleových kyselin metody MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- keratin-19 MeSH
- nádorové biomarkery MeSH
- nukleové kyseliny MeSH
BACKGROUND: Even after successful radical treatment of lung cancer, patients in stages I and II of the TNM system very frequently suffer recurrence, which end lethally. Detection of subclinical residual disease after surgery is thus one of the most important emerging diagnostic methods. Minimal residual disease (MRD) is defined as the presence of isolated tumor cells or circulating cells in a patient after curative primary tumor removal and at the same time, no clinical signs of cancer. Conventional methods cannot detect minimal residual disease and hence there is a need for detection using new molecular biological methods. METHODS: We searched the PubMed database for original and review articles on minimal residual disease in lung cancer. Search words were "lung cancer", "minimal residual disease" and "detection of minimal residual disease". The publications we found were compared with the results of our own studies on the detection of minimal residual disease in lung cancer and the personal experiences are described. Examination of blood samples from 98 healthy volunteers and bone marrow from 12 patients with non inflammatory and non tumour illness, were used to determine cut-off values for specific markers in the compartments. Subsequently, expression of selected markers in tumor tissue was analysed in a pilot sample of 50 patients with lung cancer and the presence of MRD was measured as expression of values of the tested markers correlated with clinico-pathological characteristics. CONCLUSIONS: Recent studies on other malignancies apart from lung cancer have shown the importance of MRD detection in the determination of disease progression and prognosis. The methods of MRD diagnostics are based on detection of specific tumor markers. Of these, the most specific for lung cancer, appears to be the LunX protein. The best method for determining MRD is probably RT-PCR. Further studies should expand knowledge in this area: to refine understanding of the importance of tumor markers for prognosis, as well as to confirm the significance of these findings in clinical practice.
- MeSH
- erbB receptory genetika MeSH
- fosfoproteiny genetika MeSH
- glykoproteiny genetika MeSH
- keratin-19 genetika MeSH
- lidé MeSH
- messenger RNA krev MeSH
- nádorové biomarkery krev MeSH
- nádorové cirkulující buňky * MeSH
- nádory plic krev diagnóza chirurgie MeSH
- nemalobuněčný karcinom plic krev diagnóza chirurgie MeSH
- protoonkogenní proteiny c-met genetika MeSH
- reziduální nádor MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- BPIFA1 protein, human MeSH Prohlížeč
- erbB receptory MeSH
- fosfoproteiny MeSH
- glykoproteiny MeSH
- keratin-19 MeSH
- messenger RNA MeSH
- nádorové biomarkery MeSH
- protoonkogenní proteiny c-met MeSH
The neural crest (NC) is a transient dynamic structure of ectodermal origin, found in early vertebrate embryos. The multipotential NC cells migrate along well defined routes, differentiate to various cell types including melanocytes and participate in the formation of various permanent tissues. As there is only limited information about the molecular mechanisms controlling early events in melanocyte specification and development, we exploited the AMV v-Myb transcriptional regulator, which directs differentiation of in vitro chicken NC cells to the melanocyte lineage. This activity is strictly dependent on v-Myb specifically binding to the Myb recognition DNA element (MRE). The two tamoxifen-inducible v-Myb alleles were constructed one which recognizes the MRE and one which does not. These were activated in ex ovo NC cells, and the expression profiles of resulting cells were analyzed using Affymetrix microarrays and RT-PCR. These approaches revealed up-regulation of the BMP antagonist Gremlin 2 mRNA, and down-regulation of mRNAs encoding several epithelial genes including KRT19 as very early events following the activation of melanocyte differentiation by v-Myb. The enforced v-Myb expression in neural tubes of chicken embryos resulted in detectable presence of Gremlin 2 mRNA. However, expression of Gremlin 2 in NC cells did not promote formation of melanocytes suggesting that Gremlin 2 is not the master regulator of melanocytic differentiation.
- Klíčová slova
- KRT19, Melanocyte development, PRDC, Tamoxifen-inducible v-Myb, v-Myb-dependent genes,
- MeSH
- aktivace transkripce * MeSH
- alely MeSH
- buněčná diferenciace * MeSH
- crista neuralis cytologie MeSH
- keratin-19 genetika metabolismus MeSH
- kostní morfogenetický protein 5 genetika metabolismus MeSH
- kultivované buňky MeSH
- kuřecí embryo MeSH
- melanocyty fyziologie MeSH
- mezibuněčné signální peptidy a proteiny genetika metabolismus MeSH
- onkogenní proteiny v-myb fyziologie MeSH
- ptačí proteiny genetika metabolismus MeSH
- regulace genové exprese MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- transkriptom MeSH
- zvířata MeSH
- Check Tag
- kuřecí embryo MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- keratin-19 MeSH
- kostní morfogenetický protein 5 MeSH
- mezibuněčné signální peptidy a proteiny MeSH
- onkogenní proteiny v-myb MeSH
- ptačí proteiny MeSH
