title: SMARCB1/INI1-deficient sinonasal carcinoma shows methylation of RASSF1 gene: A clinicopathological, immunohistochemical and molecular genetic study of a recently described entity.
abstract: The aim of the study was detailed clinicopathological investigation of SMARCB1/INI1-deficient sinonasal carcinomas, including molecular genetic analysis of mutational status and DNA methylation of selected protooncogenes and tumor suppressor genes by means of next generation sequencing (NGS) and methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA). A total of 4/56 (7%) cases of SMARCB1/INI1-deficient carcinomas were detected among 56 sinonasal carcinomas diagnosed over a 19year period using immunohistochemical screening. The series comprised 3 males and 1 female, aged 27-76 years (median 64 years). All tumors arose in the nasal cavity. Three neoplasms were diagnosed in advanced stage pT4. During the follow-up period (range 14-111 months (median 72 months)), three tumors recurred locally, but none of the patients developed regional or distant metastases. Ultimately, two patients died due to the tumor. Microscopically, all tumors consisted of infiltrating nests of polygonal basaloid cells with a variable component of rhabdoid cells with eosinophilic cytoplasm. Immunohistochemically, there was almost diffuse expression of cytokeratins (CK), p16, p40 and p63 in all cases, while expression of CK5/6, CK7 and vimentin was only focal or absent. The detection of NUT gave negative results. In three cases, the absence of SMARCB1/INI1 expression was due to deletion of SMARCB1/INI1 gene. Methylation of SMARCB1/INI1 gene was not found. One tumor harbored HPV18 E6/E7 mRNA. All 12 genes (BRAF, BRCA1, BRCA2, KIT, EGFR, KRAS, NRAS, PDGFRA, PIK3CA, PTEN, RET, and ROS1) tested for mutations using NGS were wild-type. Regarding DNA methylation, all four SMARCB1/INI1-deficient tumors showed methylation of RASSF1 gene by means of MS-MLPA. There was a statistically significant difference in RASSF1 gene methylation between SMARCB1/INI1-deficient and SMARCB1/INI1-positive tumors (p=0.0095). All other examined genes (ATM, BRCA1, BRCA2, CADM1, CASP8, CD44, CDKN1B, CDKN2A, CDKN2B, CHFR, DAPK1, ESR1, FHIT, GSTP1, HIC1, KLLN, MLH1a, MLH1b, RARB, and VLH) were unmethylated. In summary, we described four cases of SMARCB1/INI1-deficient sinonasal carcinoma with detailed clinicopathological data indicating that these tumors can be regarded as a distinct entity with aggressive behaviour. For the first time, we performed analysis of DNA methylation in SMARCB1/INI1-deficient sinonasal carcinomas, reporting on significantly higher methylation of RASSF1 gene in this neoplasm.
authors.affiliation: The Fingerland Department of Pathology, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia. Electronic address: lacoj@lfhk.cuni.cz.
authors.forename: Jan
authors.initials: J
authors.lastname: Laco
authors.affiliation: Institute for Clinical Biochemistry and Diagnostics, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia.
authors.forename: Marcela
authors.initials: M
authors.lastname: Chmelařová
authors.affiliation: The Fingerland Department of Pathology, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia.
authors.forename: Hana
authors.initials: H
authors.lastname: Vošmiková
authors.affiliation: The Fingerland Department of Pathology, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia.
authors.forename: Kateřina
authors.initials: K
authors.lastname: Sieglová
authors.affiliation: Institute for Clinical Biochemistry and Diagnostics, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia.
authors.forename: Ivana
authors.initials: I
authors.lastname: Bubancová
authors.affiliation: Department of Pathology, Charles University, First Faculty of Medicine and General University Hospital in Prague, Czechia.
authors.forename: Pavel
authors.initials: P
authors.lastname: Dundr
authors.affiliation: Department of Pathology, Charles University, First Faculty of Medicine and General University Hospital in Prague, Czechia.
authors.forename: Kristýna
authors.initials: K
authors.lastname: Němejcová
authors.affiliation: Department of Clinical and Molecular Pathology, Palacky University, Faculty of Medicine and Dentistry and University Hospital Olomouc, Czechia.
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Michálek
authors.affiliation: Department of Otorhinolaryngology and Head and Neck Surgery, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia.
authors.forename: Petr
authors.initials: P
authors.lastname: Čelakovský
authors.affiliation: Department of Dentistry, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia.
authors.forename: Radovan
authors.initials: R
authors.lastname: Mottl
authors.affiliation: Department of Oncology and Radiotherapy, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia.
authors.forename: Igor
authors.initials: I
authors.lastname: Sirák
authors.affiliation: Department of Oncology and Radiotherapy, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia.
authors.forename: Milan
authors.initials: M
authors.lastname: Vošmik
authors.affiliation: The Fingerland Department of Pathology, Charles University, Faculty of Medicine and University Hospital in Hradec Kralove, Czechia.
authors.forename: Aleš
authors.initials: A
authors.lastname: Ryška
mesh_terms: D000328:Adult; D000368:Aged; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D002277:Carcinoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D019175:DNA Methylation; D005260:Female; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008444:Maxillary Sinus Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008875:Middle Aged; D009367:Neoplasm Staging; D000071796:SMARCB1 Protein / Q000235:genetics* / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; C411762:RASSF1 protein, human; D000071796:SMARCB1 Protein; D025521:Tumor Suppressor Proteins
keywords: DNA methylation; RASSF1; Rhabdoid; SMARCB1/INI1; Sinonasal tract; Squamous cell carcinoma