HMGB1 genetic polymorphisms in oral squamous cell carcinoma and oral lichen planus patients.

OBJECTIVES: This study examined the single nucleotide polymorphisms (SNPs) in high-mobility group box 1 (HMGB1) gene in patients with oral squamous cell carcinoma (OSCC) and oral lichen planus (OLP). MATERIALS AND METHODS: The study was conducted on 93 patients with OSCC, 53 patients with OLP, and 100 controls, all Caucasians of the same ethnicity, matched by age. HMGB1 genotypes for 4 SNPs, 2262G/A (rs1045411), 1177G/C (rs3742305), 3814C/G (rs2249825), and rs4540927, were assessed using TaqMan SNP Genotyping Assays, Applied Biosystems. RESULTS: The HMGB1 1177GG genotype was associated with lymph-node metastasis and tumor stage in OSCCs (P = 0.016 and P = 0.030, respectively). Genotype 1177GG resulted in poorer recurrence-free survival (RFS), P = 0.000. The 1177G/C polymorphism was an independent predictor of RFS compared to GG genotype, P = 0.001. The three polymorphisms were in linkage disequilibrium (LD). The AGC and GGC haplotypes were associated with an increased oral cancer risk, determined over the haplotype odds ratios (HOR = 13.316, P = 0.015, and HOR = 5.769, P = 0.029, respectively). The AGC haplotype was related to erosive OLP progression to OSCC (HOR = 12.179, P = 0.001). CONCLUSIONS: HMGB1 polymorphism 1177G/C could be associated with tumor progression and recurrence-free survival in patients with OSCC. The haplotypes of HMGB1 gene might be associated with susceptibility to OSCC and OLP progression to OSCC.

Association of TLR2, TLR3, TLR4 and CD14 genes polymorphisms with oral cancer risk and survival.

OBJECTIVES: The aim of this study is to investigate association between polymorphisms in TLR2, TLR3, TLR4 and CD14 genes or their haplotypes with oral squamous cell carcinomas (OSCC) risk and survival. METHODS: The study was conducted on 93 OSCC patients and 104 cancer-free controls. Polymorphisms were genotyped by real-time PCR or PCR-RFLP method. RESULTS: Significant increase in oral cancer risk was observed in individuals with mutated genotype of TLR3 rs3775291 polymorphism (OR = 1.096, P = 0.036) compared to wild-type. The heterozygous and mutated genotype of TLR3 rs5743312 polymorphism had worse survival in group of patients with stage III tumours (P = 0.043). Multivariate Cox regression analysis revealed that TLR3 rs5743312 polymorphism could be considered as prognostic marker in advanced III stage OSCC (HR = 2.456, P = 0.007), but not independently of nodal status. TLR3 rs3775291 and rs5743312 polymorphisms were in strong linkage disequilibrium. Haplotype TG was associated with worse prognosis in OSCC patients in comparison with common CG haplotype (HR = 1.717, P = 0.042). Interaction among polymorphisms in TLR2, TLR3 and CD14 genes was observed (P = 0.010). CONCLUSIONS: TLR3 rs5743312 polymorphism could be considered as potential predictor of worse overall survival in advanced oral cancer, but not independently of nodal status. Haplotypes in TLR3 gene might be associated with poor prognosis in OSCC patients.

Hypermethylation of RUNX3 but not WIF1 gene and its association with stage and nodal status of tongue cancers.

OBJECTIVES: Recent studies indicate various molecular abnormalities in oral squamous cell carcinomas (OSCC), including DNA methylation of tumor-associated genes. Although promoter hypermethylation of Wnt pathway antagonists RUNX3 (Runt-related transcription factor 3) and Wnt inhibitory factor 1 (WIF1) has been identified as a common event in a number of carcinomas, methylation status and the role of RUNX3 as a possible tumor suppressor in oral and head and neck cancer are yet controversial. The aim of our study is to determine the occurrence of RUNX3 and WIF1 genes hypermethylation and correlation with tumor and host-related factors and prognosis in tongue carcinomas. MATERIAL AND METHODS: In 76 patients with tongue carcinoma, RUNX3 and WIF1 genes promoter hypermethylation analysis was assessed by nested methylation-specific PCR method. RESULTS: Hypermethylation of WIF1 and RUNX3 genes promoters was observed in 35% and 25% of carcinomas, respectively. RUNX3 gene promoter hypermethylation was significantly associated with lymph node involvement (P = 0.013) and tumor stage (P = 0.006), but not with the overall survival. Occurrence of RUNX3 and WIF1 genes comethylation was associated with nodal status (P = 0.058) and tumor stage (P = 0.055). CONCLUSIONS: Our findings indicate that RUNX3 and WIF1 are frequently aberrantly methylated and that RUNX3 promoter methylation could be considered as a potential prognostic marker in tongue carcinoma.

Interaction between the MTHFR C677T polymorphism and alcohol--impact on oral cancer risk and multiple DNA methylation of tumor-related genes.

Several studies have reported Oral Squamous Cell Carcinoma (OSCC) association with etiological factors, such as smoking and alcohol. The aim of the present study was to establish whether the methylenetetrahydrofolate reductase (MTHFR) C677T genotype and a high alcohol intake, solely or in interaction, have an impact on the oral cancer risk, DNA methylation, or multiple methylation of tumor-related genes. MTHFR C677T genetic polymorphism was determined by the PCR/RFLP method, and DNA methylation was assessed by nested methylation-specific PCR. The risk for multiple methylation was significantly increased in heavy-drinking patients with the TT genotype, compared with CC and CT patients (OR = 10.873; 95% CI, 1.134-104.24). Multiple methylation was significantly associated with tumor stage (p = 0.018), and showed a trend of association with the presence of nodal metastases (p = 0.058). A significant association was found between TT genotype and methylation status of the RASSF1A gene in OSCC patients (p = 0.012). Heavy-drinking individuals with the TT genotype showed increased oral cancer risk compared with the CC genotype (OR = 3.601; 95% CI, 1.036-12.513), and compared with the CC and CT genotypes (OR = 4.288; 95% CI, 1.325-13.877). Our study suggested gene-environment interactions between high alcohol intake and the MTHFR 677TT genotype for elevated oral cancer risk, with a significant impact on multiple methylation of cancer-related genes.

Proinflammatory cytokines (IL-1beta and TNF-alpha) and chemokines (IL-8 and MIP-1alpha) as markers of peri-implant tissue condition.

Analysis of peri-implant crevicular fluid (PICF) offers a non-invasive means of studying the host response in peri-implant disease and may provide an early indication of patients at risk for active disease. This study examined the PICF levels of interleukin-1beta (IL-1beta), tumour necrosis factor alpha (TNF-alpha), interleukin-8 (IL-8) and macrophage inflammatory protein-1alpha (MIP-1alpha) in patients with non-manifesting inflammation, early and late stages of mucositis. The study group comprised 90 adult healthy volunteers with endosseal titanium implants inserted. Samples were taken from peri-implant sulcus using a filter paper technique. Implant tissues were categorized clinically as healthy, early mucositis or advanced mucositis. Clinical manifestations were determined by: gingival index and bleeding on probing, plaque index and radiographic analyses. Cytokine concentrations were assesed using commercially available enzyme-linked immunosorbent assay kits. Patients from the control group (healthy patients) have significantly lower concentrations of IL-1beta, TNF-alpha, IL-8 and MIP-1alpha in PICF compared with both groups with mucositis. Positive correlation was noted in the control group between IL-1beta and TNF-alpha and between MIP-1alpha and IL-8 in the group with early mucositis. The results suggest that cytokines could be prognostic markers of implant failure.

Prognostic significance of TP53 mutations in oral squamous cell carcinoma with human papilloma virus infection.

PURPOSE: The aim of this study was to analyze the prognostic impact of mutated TP53 in patients with oral squamous cell carcinoma (OSCC) whose tumors were infected with human papillomavirus (HPV). METHODS: Thirty-two HPV-positive OSCC patients were included. Most of them were clinically classified as stage III (n=29). All patients underwent postoperative radiotherapy (follow-up from 12 to 60 months, median 32). There were 21 relapses. DNA was isolated by phenol extraction from tumor tissue. HPV DNA (type 16, 18, 31, 33) was detected in genomic DNA of the tumors by the PCR-PAGE method. TP53 mutations (exons 4-8) were detected by the PCR-SSCP method. RESULTS: A statistically significant difference in the number of relapses in HPV-infected (13/21) versus HPV-infected and TP53-mutated (8/8) patients was observed. Patients with both TP53 mutation and HPV infection had a significantly shorter disease-free interval than patients with HPV infection only (median 6 versus 31 months, respectively). CONCLUSIONS: TP53 mutations are associated with a higher risk of relapse and contribute to an even worse prognosis of patients with OSCC when the tumors are HPV infected. The shorter disease-free interval in patients with TP53 mutations indicates that the response to postoperative radiotherapy may be influenced by TP53 status. The presence of both HPV infection and TP53 mutations may define a particular group of tumors with a more aggressive phenotype in advanced OSCC.

[Vascularised osteocutaneous scapular flap in the reconstruction of mandibular defects after war wounding].

Vascularized osteocutaneous scapular flaps belong into the most convenient ones in reconstruction of mandibular defects followed by massive loss of adjacent skin and mucous membrane due to its vascular supply, bulkiness, suitability and mobility of cutaneous component of the flap. In seven wounded patients such defects of the mandible and adjacent soft tissues after war wounding have been reconstructed at the Clinic for Maxillofacial Surgery of the Military Medical Academy - Belgrade during the six years period with vascularized osteocutaneous scapular grafts. Here we present localization and structure of the defect, features of harvested compound graft, the procedure of reestablishment of mandibular continuity, immediate and late complications during the consolidation period, as well as the analysis of the success rate.