The main focus of the study was to detect circulating tumor cells (CTCs) in ovarian cancer (OC) patients using a new methodological approach (MetaCell(TM)) which is based on size-dependent separation of CTCs and subsequent cytomorphological evaluation. Cytomorphological evaluation using vital fluorescence microscopy approach enables to use the captured cells for further RNA/DNA analysis. The cytomorphological analysis is then completed by gene expression analysis (GEA). GEA showed that relative expression of EPCAM is elevated in CTC-enriched fractions in comparison to the whole peripheral blood sample and that the expression grows with in vitro cultivation time. Comparison of the relative gene expression level in the group of peripheral blood samples and CTC-fraction samples confirmed a statistically significant difference for the following genes (p < 0.02): KRT7, WT1, EPCAM, MUC16, MUC1, KRT18 and KRT19. Thus, we suggest that the combination of the above listed genes could confirm CTCs presence in OC patients with higher specificity than when GEA tests are performed for one marker only. The GEA revealed two separate clusters identifying patients with or without CTCs.

• Publikační typ
• časopisecké články MeSH

The major cause of death due to cancer is its metastatic deposit in numerous tissues and organs. The metastatic process requires the migration of malignant cells from primary sites to distant environments. Even for tumors initially spreading through lymphatic vessels, hematogenous transport is the most common metastatic pathway. The detachment of cancer cells from a primary tumor into the blood stream is called epithelial-mesenchymal transition (EMT). As these cells circulate further in the bloodstream they are known as circulating tumor cells (CTCs). The CTC population is highly resilient, enabling the cells to colonize a foreign microenvironment. Alternatively, cancer stem cells (CSCs) may arise from differentiated cancer cells through EMT and an embryonic transdifferentiation process. The presence of CTCs/CSCs in blood seems to be a determining factor of metastasis. This paper reviews various methods of clinical cancer detection as well as the biology and molecular characterization of CTCs/CSCs. Our goal was to summarize clinical studies which used CTC/CSCs for prognosis in patients with breast, colorectal, prostate, lung, ovarian, and bladder cancer.

• MeSH
• epitelo-mezenchymální tranzice MeSH
• lidé MeSH
• nádorové cirkulující buňky patologie   MeSH
• nádorové kmenové buňky metabolismus   MeSH
• nádory diagnóza   etiologie   mortalita   patologie   terapie   MeSH
• prognóza MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

The focus of the study was to implement a new workflow for circulating tumor cells (CTCs) characterization that would allow the analysis of CTCs on a cytomorphological and molecular level in patients with diagnosed gynecological cancer. Our findings may be useful in future cancer patient management. The study introduces a size-based enrichment (MetaCell(®)) method for the separation of viable CTCs, followed by CTCs culturing in vitro and gene expression characterization. It is based on the observation of CTCs and DTCs (Disseminated Tumor Cells) in several case studies of ovarian, endometrial and cervical cancer by means of cytomorphology and gene expression profiling. The viability of the enriched CTCs was estimated using vital and lethal fluorescence nuclear staining. This type of staining may be predictive for the success rate of subsequent CTC growth in vitro. To identify CTCs in the enriched CTC fraction, cytomorphological evaluations based on vital fluorescence staining were followed by gene expression analysis of tumor-associated (TA) genes. Cytokeratin expression (KRT7, KRT19) was analyzed in combination with MUC1, MUC16, CD24, CD44 and ALDH1. Gene expression analysis has shown that short-term in vitro culture enhanced the differentiation process of the captured CTCs growing on a membrane. On the other hand, redundant white blood cells captured on the membrane were eliminated during a short-term culture. The most frequently elevated genes in ovarian cancer (serous type) are EPCAM, KRT19 and MUC1. It has been demonstrated that CTC presence revealed by cytomorphological evaluation may be usefully complemented by TA-gene expression analysis, to increase the sensitivity of the analysis.

• Publikační typ
• časopisecké články MeSH

Characterization of molecular changes in the population of circulating tumor cells (CTCs) in the connection to the applied therapy should be the ultimate goal of the proposed research project. Molecular gene expression profiling of CTCs, in metastatic ovarian cancer patients, could be one of the possible ways, how to identify all of the individual differences in the cancerogenesis and cancer therapy process (so called “real-time” tumor biopsy). Identification of biomarkers predicting resistance on the molecular level should bring us new valid information for oncotherapy individualization.

Charakterizace molekulárních změn v populaci cirkulujících nádorových buněk (CTC) v souvislosti s aplikovanou léčbou by měla být konečným cílem navrhovaného výzkumného projektu. Molekulární profilování genové exprese CTC u metastatických pacientek s karcinomem vaječníků, by mohlo být jedním z možných způsobů, jak popsat změny v kancerogenezi v průběhu terapeutického procesu, (tzv. "real-time" nádor biopsie). Identifikace biomarkerů na molekulární úrovni predikujích rezistenci k léčivům by měla přinést nové informace pro individualizaci onkoterapie.

• MeSH
• adherence k farmakoterapii MeSH
• exprese genu MeSH
• individualizovaná medicína MeSH
• karcinogeneze MeSH
• karcinom diagnóza   terapie   MeSH
• metastázy nádorů MeSH
• nádorové biomarkery MeSH
• nádorové cirkulující buňky MeSH
• nádory vaječníků diagnóza   terapie   MeSH
• ženy MeSH

• Konspekt
• Gynekologie. Porodnictví
• NLK Obory
• gynekologie a porodnictví
• onkologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR