Tumour microenvironment (TME) can activate protecting signalling pathways in tumour cells, especially when TME cells are damaged by chemotherapy. In this project, we will focus on cisplatin resistance conferred to cancer cells by TME. FaDu cells will be co-cultured with primary cell lines derived from tumour tissue of individual head and neck cancer (HNSCC) patients. Effect of co-culture on resistance, invasiveness, and migratory capacity of cancer cells will be assessed. Based on those, we will propose a protocol for detection of tumour-supporting stroma in individual HNSCC patients, which is an important step in personalized medicine of HNSCC. Group of cultures well-supporting and non-supporting chemoresistance will be then analysed for differences in cell stiffness (assessed by atomic force microscopy) and for changes in gene expression (microarray technology) and amino acid profiles (ion-exchange liquid chromatograph). The main aim is to select reliable markers of tumour-supporting stroma detectable in biological samples of HNSCC patients.

Buňky v mikroprostředí nádorů (TME) mohou aktivovat v nádorových buňkách rezistenci a to zejména pokud jsou poškozeny chemoterapií. V tomto projektu se zaměříme na odolnost proti cisplatině, kterou TME uděluje nádorovým buňkám. Buněčná linie FaDu bude kultivována s primárními buněčnými liniemi pocházejícími z nádorové tkáně jednotlivých pacientů s karcinomem hlavy a krku (HNSCC). Bude posuzován vliv kokultivace na rezistenci, invazivitu a migrační kapacitu nádorových buněk. Na základě těchto experimentů navrhneme protokol pro detekci „podporujícího“ nádorového stromatu u jednotlivých HNSCC pacientů, což je důležitý krok v personalizované medicíně HNSCC. U skupin primárních buněčných linií dobře podporujících/nepodporujících chemorezistenci bude poté provedena analýzy buněčné tuhosti (mikroskopie atomárních sil), analýza genové exprese (technologie mikroarray) a změn v profilech aminokyselin (ionexová chromatografie). Hlavním cílem je vybrat spolehlivé markery přítomnosti stromatu podporujícího nádor, které by mohly být detekovány v biologických vzorcích pacientů s HNSCC.

• Klíčová slova
• biomarkery, biomarkers, Spinocelulární karcinomy v oblasti hlavy a krku, mikroprostředí nádoru, stroma podporující růst nádoru, chemorezistence, personalizovaná medicína, kokultivace, technologie microarray, Head and neck squamous cell carcinoma, tumour microenvironment, tumour-supporting stroma, chemoresistance, personalized medicine, co-culture, microarray technology,

• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Head and neck squamous cell carcinomas (HNSCC) belong among severe and highly complex malignant diseases showing a high level of heterogeneity and consequently also a variance in therapeutic response, regardless of clinical stage. Our study implies that the progression of HNSCC may be supported by cancer-associated fibroblasts (CAFs) in the tumour microenvironment (TME) and the heterogeneity of this disease may lie in the level of cooperation between CAFs and epithelial cancer cells, as communication between CAFs and epithelial cancer cells seems to be a key factor for the sustained growth of the tumour mass. In this study, we investigated how CAFs derived from tumours of different mRNA subtypes influence the proliferation of cancer cells and their metabolic and biomechanical reprogramming. We also investigated the clinicopathological significance of the expression of these metabolism-related genes in tissue samples of HNSCC patients to identify a possible gene signature typical for HNSCC progression. We found that the right kind of cooperation between cancer cells and CAFs is needed for tumour growth and progression, and only specific mRNA subtypes can support the growth of primary cancer cells or metastases. Specifically, during coculture, cancer cell colony supporting effect and effect of CAFs on cell stiffness of cancer cells are driven by the mRNA subtype of the tumour from which the CAFs are derived. The degree of colony-forming support is reflected in cancer cell glycolysis levels and lactate shuttle-related transporters.

• Publikační typ
• časopisecké články MeSH

Cancer-associated fibroblasts (CAFs) are one of the most abundant and critical components of the tumor stroma. CAFs can impact many important steps of cancerogenesis and may also influence treatment resistance. Some of these effects need the direct contact of CAFs and cancer cells, while some involve paracrine signals. In this study, we investigated the ability of head and neck squamous cell carcinomas (HNSCC) patient-derived CAFs to promote or inhibit the colony-forming ability of HNSCC cells. The effect of cisplatin on this promoting or inhibiting influence was also studied. The subsequent analysis focused on changes in the expression of genes associated with cancer progression. We found that cisplatin response in model HNSCC cancer cells was modified by coculture with CAFs, was CAF-specific, and different patient-derived CAFs had a different "sensitizing ratio". Increased expression of VEGFA, PGE2S, COX2, EGFR, and NANOG in cancer cells was characteristic for the increase of resistance. On the other hand, CCL2 expression was associated with sensitizing effect. Significantly higher amounts of cisplatin were found in CAFs derived from patients who subsequently experienced a recurrence. In conclusion, our results showed that CAFs could promote and/or inhibit colony-forming capability and cisplatin resistance in HNSCC cells via paracrine effects and subsequent changes in gene expression of cancer-associated genes in cancer cells.

• MeSH
• antitumorózní látky farmakologie   MeSH
• chemorezistence účinky léků   MeSH
• cisplatina farmakologie   MeSH
• dlaždicobuněčné karcinomy hlavy a krku metabolismus   patologie   MeSH
• fibroblasty asociované s nádorem účinky léků   metabolismus   MeSH
• kokultivační techniky MeSH
• lidé středního věku MeSH
• lidé MeSH
• lokální recidiva nádoru metabolismus   patologie   MeSH
• nádorové buněčné linie MeSH
• nádory hlavy a krku metabolismus   patologie   MeSH
• parakrinní signalizace účinky léků   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• testy nádorových kmenových buněk MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

PURPOSE: The treatment strategy of parotid gland tumours depends mainly on the histopathological type of the lesion. Fine-needle aspiration biopsy (FNAB) is recommended in preoperative diagnostics. The aim of the study was to evaluate the FNAB standing in the diagnostic algorithm of parotid gland lesions and to correlate FNAB results in relation to the definitive histopathological diagnosis. MATERIAL AND METHODS: The retrospective analyses of 651 examined and consequently surgically treated parotid gland lesions at the Department of Otorhinolaryngology and Head and Neck Surgery, 1st Faculty of Medicine, Charles University in Prague between 2006 and 2016 were used. Preoperative cytological results were consequently evaluated in relation to the definitive histopathological diagnosis. RESULTS: The cohort consisted of 367 women and 284 men (average age 58 years). FNAB was diagnostic in 604 (92.8%) patients and non-diagnostic in 47 (7.2%) patients. The result of FNAB was positive (suspicious for malignant tumour) in 89 (14.7%) patients and negative (benign) in 515 (85.3%) patients. Sensitivity of the examination was 80.00%, specificity was 93.82%, PPV 62.92%, NPV 97.28%, and LR + and LR- were 12.95 and 0.21, respectively, with an accuracy of 92.22%. CONCLUSION: Our results confirm the significant role of FNAB in the diagnostic algorithm of parotid gland lesions.

• MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory příušní žlázy * diagnóza   chirurgie   MeSH
• parotis * chirurgie   MeSH
• retrospektivní studie MeSH
• senzitivita a specificita MeSH
• tenkojehlová biopsie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH