Chronically inflamed mucosa in inflammatory bowel disease (IBD) is associated with an increased risk of cancer. Besides IBD-associated dysplasia, there are non-conventional mucosal changes that may act as potential precursors. The aim of the study was to retrospectively review samples from IBD patients focusing on detection of such lesions with evaluation of their immunohistochemical and molecular properties. Surgical specimens and/or endoscopical biopsy samples of IBD patients examined during a 10-year period were reviewed. Detected mucosal lesions were divided into three groups-group 1 (non-conventional or putative precursor lesions - PPLs) with serrated or villous hypermucinous morphology, group 2 (true serrated polyps fulfilling valid criteria), and group 3 (IBD-associated neoplasia). Lesions from all groups were analyzed with antibodies against MLH1, p53, and MGMT and by molecular testing of KRAS, NRAS, and BRAF mutation. Samples from 309 IBD patients were reviewed. A total of 88 mucosal lesions were found in 51 patients. Most common were lesions from group 1 with superficial serrated epithelial change seen in 41 samples (46.6%) and villous hypermucinous change in 6 (6.8%). Group 2 consisted of 15 true serrated polyps. Six conventional IBD-dysplasia cases and 11 carcinomas were seen in group 3. Six lesions from group 1 were associated with invasive carcinoma whereas two shared the same mutation in KRAS or BRAF. Lesions from group 1 were characterized by loss of MGMT expression in 44.6%, aberrant p53 expression, and by mutations in KRAS gene in 42.9% of cases. This study proves the existence of mucosal changes other than conventional IBD-dysplasia and extends the knowledge about their immunohistochemical and molecular properties and relation to carcinoma further supporting their potential role in IBD-related carcinogenesis.
- MeSH
- adenokarcinom patologie MeSH
- adenom diagnóza patologie MeSH
- dítě MeSH
- dospělí MeSH
- idiopatické střevní záněty diagnóza patologie MeSH
- kolorektální nádory diagnóza patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mutace genetika MeSH
- nádorové biomarkery analýza MeSH
- polypy tlustého střeva diagnóza patologie MeSH
- protoonkogenní proteiny B-raf genetika MeSH
- retrospektivní studie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Sinonasal carcinomas are head and neck tumours arising from the nasal cavity and paranasal sinuses characterized by unfavourable outcome, difficult treatment, diagnosis and prognosis. MicroRNAs are key molecules in the regulation of development and progression of cancer and their expression profiles could be used as prognostic biomarkers, to predict the patients' survival and response to treatment. In this study, we used quantitative real‑time PCR with TaqMan® Advanced miRNA Assays to investigate the relative expression values of selected micro- RNAs in a unique set of formalin-fixed paraffin-embedded tissue samples obtained from 46 patients with sinonasal squamous cell carcinoma. Our results showed statistically significant up-regulation of three mature microRNAs: miR-9-5p (fold change: 6.80), miR-9-3p (fold change: 3.07) and let-7d (fold change: 3.93) in sinonasal carcinoma patients. Kaplan-Meier survival analysis and logrank test identified association between higher expression of miR-9-5p and longer survival of the patients (P = 0.0264). Lower expression of let-7d was detected in the patients with impaired survival, and higher expression of miR-137 was linked to shorter survival of the patients. We also identified several correlations between expression of the studied microRNAs and recorded clinicopathological data. Higher expression of miR-137 and lower expression of let-7d correlated with local recurrence (P = 0.045 and P = 0.025); lower expression of miR-9-5p and higher expression of miR-155-5p correlated with regional recurrence (P = 0.045 and P = 0.036). Higher expression of miR-9-3p correlated with occupational risk (P = 0.031), presence of vascular invasion (P = 0.013) and perineural invasion (P = 0.031). Higher expression of miR-155-5p was present in the samples originating from maxillary sinus (P = 0.011), cN1-3 classified tumours (P = 0.009) and G2-3 classified tumours (P = 0.017). In conclusion, our study supports the hypothesis of future prospect to use expression of miRNAs as prognostic biomarkers of squamous cell sinonasal carcinoma. In particular, miR-9-5p and miR-9-3p seem to be important members of the sinonasal cancer pathogenesis.
- MeSH
- Kaplanův-Meierův odhad MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikro RNA genetika metabolismus MeSH
- nádorové biomarkery genetika metabolismus MeSH
- nádory sinu maxillaris genetika patologie MeSH
- prognóza MeSH
- regresní analýza MeSH
- regulace genové exprese u nádorů * MeSH
- spinocelulární karcinom genetika patologie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Cíl studie: V rámci této studie jsme využili metody kvantitativní real-time PCR pro sledování úrovně relativní exprese miR-145-5p, miR-484 a miR-99a-5p v souboru HPV pozitivních a HPV negativních vzorků sinonasálního dlaždicobuněčného karcinomu. Typ studie: původní práce.Název a sídlo pracoviště: Ústav klinické biochemie a diagnostiky, Lékařská fakulta v Hradci Králové, Univerzita Karlova a Fakultní nemocnice Hradec Králové, Sokolská 581, 500 05 Hradec Králové.Materiál a metody: Metoda kvantitativní real-time PCR s TaqManTM Advanced miRNA Assays byla použita pro sledování relativní exprese vybraných mikroRNA (miR-145-5p, miR-484 a miR-99a-5p) v unikátním souboru vzorků fixovaných ve formalínu a archivovaných v parafinu získaných od 46 pacientů se sinonasálním dlaždicobuněčných karcinomem. Statistická analýza (Studentův t-test, jednofaktorová analýza rozptylu a regresní analýza) byla provedena s cílem porovnat úroveň relativní exprese mikroRNA se zaznamenanými klinickopatologickými daty jako je HPV status.Výsledky: Naše výsledky ukazují statisticky významnou downregulaci miR-145-5p (P < 0,001 a Fold change = -2,78) ve vzorcích sinonasálního skvamózního karcinomu. Přítomnost HPV infekce korelovala s mírou exprese všech tří studovaných mikroRNA. Exprese miR-145-5p byla nižší u HPV negativních vzorků (P = 0,019), miR-99a-5p byla upregulována u HPV pozitivních vzorků (P = 0,058) a miR-484 byla downregulována u HPV pozitivních vzorků (P = 0,016). Pacienti, kteří byli zařazeni do kategorie kuřáci nebo bývalí kuřáci, vykazovali downregulaci miR-99a-5p (P = 0,060). Perineurální šíření bylo spojeno s významnou downregulací miR-99a-5p (P = 0,0055). Významná downregulace miR-145-5p byla spojena s angioinvazí (P = 0,037) a regionálním šířením (P = 0,076).Závěr: V rámci naší studie jsme nalezli spojitost mezi expresí studovaných mikroRNA a přítomností HPV infekce v nádorových vzorcích. Naše výsledky naznačují, že miR-145-5p, miR-484 a miR-99a jsou součástí patogeneze HPV. Avšak přesná role miRNA spojených s HPV infekcí ve vývoji sinonasálních karcinomů není dosud známá.
Objective: In the present study, we used quantitative real-time PCR to investigate relative expression values of miR-145-5p, miR-484 and miR-99a-5p in HPV positive and HPV negative samples of sinonasal squamous cell carcinoma. Design: Original Article. Settings: Institute of Clinical Biochemistry and Diagnostics, Charles University, Faculty of Medicine in Hradec Králové and University Hospital Hradec Králové, Sokolská 581, 500 05 Hradec Králové Materials and Methods: Quantitative real-time PCR with TaqManTM Advanced miRNA Assays was used to investigate relative expression values of selected microRNAs (miR-145-5p, miR-484 and miR-99a-5p) in a unique set of formalin-fixed paraffin-embedded tissue samples obtained from 46 patients with sinonasal squamous cell carcinoma. Statistical analysis (Student‘s t-test, one-way analysis of variance and regression analysis) was performed to compare relative expression miRNA values and recorded clinicopathological data such as HPV status. Results: Our results show statistically significant downregulation of miR-145-5p (P < 0.001 and Fold change = -2.78) in sinonasal squamous cell carcinoma samples. The presence of HPV infection correlated with the expression levels of all three studied miRNAs. The expression of miR-145-5p was lower in HPV negative samples (P = 0.019), miR-99a-5p was upregulated in HPV positive samples (P = 0.058) and miR-484 was downregulated in HPV positive samples (P = 0.016). Patients with recorded history of smoking or currents smokers showed downregulation of miR-99a-5p (P = 0.060). Perineural invasion was linked to significant downregulation of miR-99a-5p (P = 0.0055). Distinctive downregulation of miR-145-5p was linked to vascular invasion (P = 0.037) and regional recurrence (P = 0.076). Conclusion: We have found correlation between studied miRNA expression and presence of HPV infection in the patient samples. Our results suggest that miR-145, miR-484 and miR-99a are involved in HPV pathogenesis. However, the actual pathway of HPV related miRNA involvement in sinonasal tumor development is not yet known.
- Klíčová slova
- sinonasální karcinom,
- MeSH
- dlaždicobuněčné karcinomy hlavy a krku * patologie virologie MeSH
- infekce papilomavirem komplikace MeSH
- klinická studie jako téma MeSH
- lidé MeSH
- mikro RNA MeSH
- nádory vedlejších dutin nosních patologie virologie MeSH
- Papillomaviridae * izolace a purifikace MeSH
- Check Tag
- lidé MeSH
BACKGROUND: Patients with inflammatory bowel disease (IBD) - ulcerative colitis (UC) and Crohn's disease (CD) have an elevated risk of developing colorectal carcinoma (CRC). Major risk factor in IBD patients is the continuous chronic inflammation leading to development of dysplasia and carcinoma. Nevertheless, other types of non-conventional but suspicious mucosal changes serrated change/dysplasia, NOS and villous hypermucinous change, have also been reported in IBD patients. Preneoplastic potential of these lesions is still not well elucidated. AIMS: The aim of this study was identification of IBD-associated CRCs focusing on finding related precursor lesions in the surgical specimen or in archival biopsy samples followed by a detailed morphological, immunohistochemical and molecular evaluation. For the purpose of the study the mucosal lesions were divided into conventional IBD-associated dysplasia and non-conventional lesions that were merged under a provisory term of putative preneoplastic lesions (PPL). METHODS: A total of 309 consecutive IBD colectomy specimens diagnosed during a 10-year period were reviewed. Detailed morphological evaluation, immunohistochemical analysis of mismatch repair (MMR) proteins, p53 and O6-methylguanine DNA methyltransferase (MGMT) expression and molecular analysis for KRAS, NRAS and BRAF gene mutation were performed in the retrieved CRC cases as well as in the detected dysplasia and PPLs of these patients. RESULTS: We identified 11 cases of morphologically heterogenous IBD-associated CRCs, occurring in 5 males and 6 females, aged 26-79 years (mean 44 years). A total of 22 mucosal lesions were revealed in 8 CRC patients comprising conventional IBD-associated dysplasia (4 lesions), PPLs as serrated change/dysplasia NOS (11 lesions), villous hypermucinous change (5 lesions), and two true serrated lesions (one sessile serrated adenoma and one traditional serrated adenoma). More than one type of lesion was found in 6 patients. Seven CRC cases harbored mutation of KRAS/NRAS and one case of BRAF. Two patients with KRAS-mutated CRC showed the same mutation in PPL in the same specimen (one serrated change NOS and one TSA with high-grade dysplasia). Similarly, one BRAF-mutated carcinoma case presented the same mutation in serrated change/dysplasia, NOS in the same specimen. Of the CRCs, two showed deficient MMR system profile, six presented with loss of MGMT expression, and six showed aberrant p53 expression. PPLs showed deficient MGMT expression (14 cases) and aberrant p53 (10 cases) as well. CONCLUSION: IBD-associated CRCs are very heterogeneous entities. Besides conventional IBD-related dysplasia, other types of mucosal lesions may be associated with long lasting IBD and CRC e.g. villous hypermucinous change and serrated change/dysplasia, NOS. Since these lesions share certain genetic or immunohistochemical changes with the related CRC, a suspicion is raised that these lesions may also have preneoplastic potential. Awareness of these changes is necessary to prevent their missing and under-reporting, and further studies of these lesions should be carried out.
- MeSH
- adenokarcinom etiologie metabolismus patologie MeSH
- dospělí MeSH
- idiopatické střevní záněty komplikace metabolismus patologie MeSH
- imunohistochemie MeSH
- kolorektální nádory etiologie metabolismus patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- střevní sliznice metabolismus patologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
