Ruzica Kozomara

Prognostic significance of tumor-related genes hypermethylation detected in cancer-free surgical margins of oral squamous cell carcinomas

Oral squamous cell carcinoma (OSCC) is characterized by high mortality rates. High incidence of local recurrences in the normal-appearing surgical margins of OSCC patients indicates the existence molecular alterations, including DNA methylation, which could not be detectable by histopathologic analysis. The objective of our study was to determine correlation of tumor-related genes hypermethylation detected in histopathologically negative surgical margins with clinical and prognostic parameters. The genes selected for methylation analysis covered a wide range cellular processes including cell cycle control (p16), apoptosis (DAPK and RASSF1A), Wnt signaling (APC, WIF1 and RUNX3), cell-cell adhesion (E-cad), and DNA repair (MGMT and hMLH1). All of 47 patients had histologically confirmed negative surgical margins. For each of patient, samples from primary malignant tissue and the two consecutive surgical margins were taken at the time of surgery. DNA methylation was determined by multiplex nested methylation-specific PCR. Twenty-seven patients were margin-positive for promoter hypermethylation of at least one gene under study. The presence of DAPK promoter hypermethylation detected in surgical margins was associated with the decreased overall survival (p=0.004, log rank test). Multivariate analysis revealed that DAPK promoter hypermethylation in surgical margins is an independent prognostic factor for overall survival, HR=4.105 (1.458-11.555, 95% CI, p=0.007). Hypermethylation of other tumor-related genes under study did not have prognostic significance. These results demonstrate that DNA hypermethylation in histologically negative surgical margins is a frequent event. Promoter hypermethylation of DAPK gene detected in surgical margins may be a useful molecular marker for the poor survival in OSCC patients. Further investigation into the therapeutic potential of these findings in OSCC is warranted.

Vitamin D receptor, CYP27B1 and CYP24A1 genes polymorphisms association with oral cancer risk and survival.

BACKGROUND Genetic polymorphisms of vitamin D receptor gene (VDR) and genes involved in vitamin D metabolism pathway, CYP27B1 and CYP24B1, may affect individual susceptibility to oral squamous cell carcinoma. The purpose of this study was to evaluate the associations between VDR, CYP27B1 and CYP24A1 gene polymorphisms with oral cancer risk and survival. METHODS Study cohort consisted of 110 patients with oral cancer and 122 healthy controls. The genotypes of the analysed genes were determined by PCR-RFLP or real-time PCR method. RESULTS The significant decrease of oral cancer risk was observed in individuals with heterozygote genotype of CYP24A1 gene (rs2296241) (odds ratio 0.281, P = 0.000) in comparison with wild type. Patients with VDR FokI ff wild type genotype had significantly worse overall survival (P = 0.012, log rank) compared with heterozygous and mutated genotype combined. A stratified analysis by the lymph node involvement and tumour stage showed that ff is associated with poor survival in groups with and without lymph node involvement (P = 0.025, P = 0.040, respectively) and in stage III tumours (P = 0.026). Multivariate Coxs regression analysis revealed that VDR FokI could be considered an independent prognostic factor. CONCLUSION Our findings indicate that CYP24A1 gene polymorphism might have an influence on the susceptibility to oral cancer. VDR FokI polymorphism was associated with worse survival and could be considered as an independent prognostic marker.

Association of VEGF-A genetic polymorphisms with cancer risk and survival in advanced-stage oral squamous cell carcinoma patients

BACKGROUND Vascular endothelial growth factor (VEGF), a key mediator of angiogenesis, is overexpressed in a wide variety of human cancers, including oral squamous cell carcinoma (OSCC). In this study, we examined whether individual polymorphisms within VEGF-A gene, rs699947 (-2578C/A), rs1570360 (-1154G/A), rs2010963 (-634G/C), rs3025039 (+936C/T) or their haplotypes are associated with an oral cancer risk and survival. METHODS A case-control study was conducted on 114 OSCC patients and control group of 126 individuals without a previous cancer history, all the Caucasian race and the same ethnicity, matched by age and gender. VEGF-A genotypes were analyzed using TaqMan SNP Genotyping Assays, Applied Biosystems. RESULTS The -1154 GG genotype was significantly associated with the decreased overall survival in OSCC patients (p = 0.010, log rank test). Stratified analysis revealed that in patients with nodal metastases and stage III, -1154 GG genotype was related to worse survival, p = 0.009, p = 0.013, respectively. The multivariate analysis revealed that -1154 GG genotype is an independent adverse factor for survival in the OSCC (HR = 1.899, [1.138-3.168], p = 0.014). The +936 CC genotype was associated with advanced staged OSCC (p = 0.050). The three polymorphisms, -2578, -1154 and -634 were in linkage disequilibrium (LD). The CAG haplotype could be associated with an increased oral cancer risk, OR = 7.967, [1.730-36.689], p = 0.008, while CGG haplotype could be associated with a decreased oral cancer risk, OR = 0.561, [0.326-0.964], p = 0.036. CONCLUSIONS VEGF-A -1154 GG genotype could be considered as a prognostic marker of poor survival in advanced-stage OSCC patients. Haplotypes of VEGF-A gene may be associated with susceptibility to OSCC.

Genetic Polymorphisms of ADH1C and CYP2E1 and Risk of Oral Squamous Cell Carcinoma

Objective. Several studies have suggested that the metabolism of alcohol is modulated by the polymorphisms in genes encoding ethanol-metabolizing enzymes, including alcohol dehydrogenase 1C, ADH1C, and cytochrome P450-dependent monooxygenase, CYP2E1. Genetic polymorphisms of ethanol-metabolizing enzymes may affect individual susceptibility to oral cancer. The purpose of this study was to investigate the associations between ADH1C and CYP2E1 gene polymorphisms with oral squamous cell carcinoma in an ethnically homogeneous Caucasian population. Design. Case-control study. Setting. Serbian national general hospital. Subjects and Methods. The study was conducted on 123 oral cancer patients and a control group of 177 individuals of the Caucasian race and the same ethnicity, matched in age and gender, without previous cancer history. The control group consisted of 120 population-based and 57 hospital-based controls of heavy-drinking individuals. Genetic polymorphisms of ADH1C SspI, ADH1C HaeIII, CYP2E1 RsaI, and CYP2E1 Ins were determined by the polymerase chain reaction and restriction fragment length polymorphisms. Results. After adjustment by potential confounders, the significant increase of oral cancer risk, independent of alcohol drinking, was observed in individuals with the variant ADH1C SspI*2/*2 genotype (odds ratio, 3.029; P = .014) and in combined ADH1C SspI*1/*2 and ADH1C SspI*2/*2 genotypes (odds ratio, 2.605; P = .002), compared to the ADH1C*1/1* wild type. The association of other polymorphisms under study was not observed. Conclusion. This study suggested that the ADH1C SspI polymorphism could play a significant role in the etiology of oral cancer, whereas ADH1C HaeIII, CYP2E1 RsaI, and CYP2E1 Ins could have minor influence.

Concha bullosa and the nasal middle meatus obstructive syndrome

BACKGROUND Concha bullosa (CB) is pneumatization of the middle turbinate and one of the most common anatomic variation of the sinonasal region. It is found in about 25% of the population. Middle meatus obstructive syndrome (MMOS) is, usually connected with CB. The main symptoms of this syndrome are headaches, impaired nasal breathing and hyposmia. Headache is the most common symptom and it may occur due to contact between a CB and other structures of the nasal cavity. CASE REPORT We presented a case of 32 year-old-woman with headaches, located in the orbital and the left frontal region. The headaches were intermittent and corresponding to the nasal cycle. After neurologic and allergic examination, endoscopic nasal examination demonstrated a septal deviation to the right side and a large middle turbinate in the left side of the nasal cavity. Coronal computerized tomography (CT) of the paranasal sinuses demonstrated the septal deformation and pneumatization of the left middle turbinate. Diagnosis was confirmed by lidocaine test. In the functional endoscopic surgery (FESS), the lateral lamela of the anterior CB was removed. At the same time, the septoplasty was done. At the control examination, the patient was without symptoms. CONCLUSION Although CB is the common anatomic variation of the nasal cavity, MMOS is rare. Headache (rhinogenic origin) is the most important symptom. Surgical treatment is the lateral resection of the CB in the FESS technique and the septoplasty.

Genetic polymorphisms of catechol-o-methyltransferase: Association with temporomandibular disorders and postoperative pain

AIMS To evaluate the association between catechol-O-methyltransferase (COMT) gene polymorphisms and temporomandibular disorders (TMD), TMD pain, psychosocial impairment related to TMD, and postoperative pain. METHODS A total of 90 patients with a diagnosis of painful TMD and 92 matched controls were investigated for the presence of TMD, TMD pain, and psychosocial variables by the Research Diagnostic Criteria for TMD. In a prospective cohort study of 40 subjects who underwent extraction of at least one fully impacted mandibular third molar, subjects had 6 months post-surgery follow-up of postoperative pain. DNA extracted from peripheral blood was genotyped for three COMT polymorphisms (rs4680, rs6269, and rs165774) by real-time TaqMan method. The association between COMT polymorphisms and clinical variables was determined by calculating odds ratios (OR) and their 95% confidence intervals (CI). RESULTS Homozygous AA genotype and heterozygous variant A allele carriers (genotype AG/AA) for rs165774 polymorphism were associated with increased risk of TMD compared to wild type (wt) GG genotype (OR = 9.448, P = .006; OR = 2.088, P = .017, respectively). In addition, AA genotype was associated with increased risk of arthralgia (OR = 4.448, P = .011), myofascial pain (OR = 3.543, P = .035), and chronic TMD pain (OR = 6.173, P = .006), compared to wt genotype. AA genotype for rs6269 polymorphism was related to less postoperative chronic TMD pain (P = .025) and lower postoperative acute pain at the extraction site (P = .030). No associations with depression and somatization were observed. CONCLUSION AA genotype of rs165774 could be a significant risk factor for the development of TMD and TMD pain, while AA genotype of rs6269 presents less postoperative chronic TMD pain and acute pain at a dental extraction site.

Mucocoele of the maxillary sinus

BACKGROUND Mucocoele is histopathologically benign, cystic change of paranasal sinuses filled with mucoid contents, which with its growth is pressuring and destroying local bone walls. In only 3% of the cases it can be localized in maxillary sinuses. Etiology is unknown. Pyocoele develops by secondary infection. CASE REPORT The male patient was 21 years old. His symptoms were runny nose with thick contents and heavy breathing on the right side of the nose, headaches, as well as the swelling of the right cheek. During clinical examination, the expansive change was found. It was completely closing the right side of the nose cavity. Computerized tomography (CT) of paranasal cavities showed excessive expansion of the right maxillary sinus, with very thinned walls, while the cavity was filled with liquid. After antibiotics therapy, the radical operation of the right maxillary sinus was performed, based on Caldwell Luc method. The frontal wall was found to be extremely convex and thinned, while the medial wall was with dehiscention. The cystic change was extirpated. Mucocoele was proved by pathohistologic findings. Its wall was about 2 mm thick and it showed squamous metaplasia in the large part of the mucocoela epithel. In the submucosa fibrosis and inflammatory infiltrate was present. Postoperative follow-up was under control. Clinicaly and radiographycaly, six months after therapy, the patient does not have troubles. CONCLUSION The rare localization of the mucocoeles in maxillary sinus can be explained with the width of the maxillary ostia. Infected mucocoeles, expanded to the local anatomical structures, should be operated on with classic radical surgical operation.

Pulp Sensitivity: Influence of Sex, Psychosocial Variables, COMT Gene, and Chronic Facial Pain

Introduction: The purpose of this study was to evaluate the associations of variability in pulp sensitivity with sex, psychosocial variables, the gene that encodes for the enzyme catechol‐O‐methyltransferase (COMT), and chronic painful conditions (temporomandibular disorders [TMDs]). Methods: The study was composed of 97 subjects (68 women and 29 men aged 20–44 years). The electric (electric pulp tester) and cold (refrigerant spray) stimuli were performed on mandibular lateral incisors. The results were expressed as pain threshold values for electric pulp stimulation (0–80 units) and as pain intensity scores (visual numeric scale from 0–10) for cold stimulation. The Research Diagnostic Criteria for TMD were used to assess TMD, depression, and somatization. DNA extracted from peripheral blood was genotyped for 3 COMT polymorphisms (rs4680, rs6269, and rs165774) using the real‐time TaqMan method. Multivariate linear regression was used to investigate the joint effect of the predictor variables (clinical and genetic) on pulp sensitivity (dependent variables). Results: Threshold responses to electric stimuli were related to female sex (P < .01) and the homozygous GG genotype for the rs165774 polymorphism (P < .05). Pain intensity to cold stimuli was higher in TMD patients (P < .01) and tended to be higher in women. Multivariate linear regression identified sex and the rs165774 COMT polymorphism as the determinants of electric pain sensitivity, whereas TMD accounts for the variability in the cold response. Conclusions: Our findings indicate that sex/a COMT gene variant and TMD as a chronic painful condition may contribute to individual variation in electric and cold pulp sensitivity, respectively.