Daniela Mielcarek-Kuchta

The negative regulators of Wnt pathway—DACH1, DKK1, and WIF1 are methylated in oral and oropharyngeal cancer and WIF1 methylation predicts shorter survival

The deregulation of Wnt signaling has recently emerged as one of the drivers of head and neck cancers. This is frequently related to the methylation of several antagonists of this pathway. This study aimed at the assessment of the profile of methylation of Wnt pathway antagonists and the determination of the prognostic value of the methylation of selected genes in oral carcinomas. The methylation of DACH1, DKK1, LKB1, PPP2R2B, RUNX3, SFRP2, and WIF-1 was analyzed in 16 oral squamous cell carcinoma cell lines using the methylation-specific polymerase chain reaction. The methylation of selected genes was further analyzed in tumor sections from 43 primary oral carcinoma patients. The analysis of oral carcinoma cell lines showed very frequent methylation of SFRP2 and WIF-1 and also a less frequent methylation of DACH1 and DKK1. On the other hand, RUNX3 was methylated only in one cell line, while LKB1 and PPP2R2B were not methylated in any of the cell lines. The biallelic methylation of DKK1 correlated with the low level of expression of this gene. Further evaluation of the methylation of DACH1, DKK1, and WIF1 in a clinical patient group confirmed the frequent methylation of WIF1 and intermediate or low frequency of methylation of DACH1 or DKK1, respectively. Importantly, the methylation of WIF-1 correlated with shorter survival in oral cancer patients. Overall, the methylation of the antagonists of Wnt pathway is frequently detected in oral squamous cell carcinomas. The methylation of WIF1 may be considered a prognostic marker in oral cancers.

Prognostic significance of the methylation of Wnt pathway antagonists—CXXC4, DACT2, and the inhibitors of sonic hedgehog signaling—ZIC1, ZIC4, and HHIP in head and neck squamous cell carcinomas

ObjectivesAberrations in Wnt and Shh signaling pathways are related to the pathogenesis of head and neck carcinomas, and their activation frequently results from epigenetic alterations. This study aimed to assess the frequency of methylation of negative regulators of Wnt signaling: CXXC4, DACT2, HDPR1, and FBXW11 and Shh signaling: HHIP, PTCH1, SUFU, ZIC1, and ZIC4 and correlate it with clinicopathological features in this group of patients.Materials and methodsMethylation-specific PCR was used to detect gene promoter methylation, and real-time PCR was used to assess gene expression level.ResultsThe analysis of the occurrence of gene promoter methylation in head and neck carcinoma cell lines indicated that CXXC4, DACT2, HHIP, ZIC1, and ZIC4 are methylated in these tumors. These genes were further analyzed in tumor sections from oral and laryngeal cancer patients. Gene methylation rate was higher in laryngeal tumors. The methylation index in tumor samples correlated with the overall survival in a subgroup of oral cancer patients who died of the disease. Moreover, ZIC4 methylation correlated with lymph node involvement in oral cancer patients.ConclusionsOur findings corroborate that the activation of Wnt signaling in head and neck squamous cell carcinoma (HNSCC) is related to epigenetic silencing of its negative regulators. Moreover, the results indicate that the same mechanism of activation may operate in the case of Shh signaling.Clinical relevanceThe methylation of ZIC4 may be considered a new prognostic marker in oral cavity and oropharyngeal tumors. Further investigations should determine the diagnostic significance of methylation of ZIC4, HHIP, and DACT2 in head and neck carcinomas.

Prognostic factors in oral and oropharyngeal cancer based on ultrastructural analysis and DNA methylation of the tumor and surgical margin

Oral and oropharyngeal cancers are characterized by relatively low 5- year survival rates due to many factors, including local recurrence. The identification of new molecular markers may serve for the estimation of prognosis and thus augment treatment decisions and affect therapy outcome. The aim of this study was to describe the morphological characteristics and the DNA methylation status of the CDKN2A,CDH1, ATM, FHIT and RAR- genes in the central and peripheral part of the tumor and the surgical margin and evaluate their prognostic significance. 53 patients with oral and oropharyngeal cancer were enrolled to the prospective study, and had been primarily treated surgically. Correlations between morphological data, hypermethylation status and clinicopathological data, as well as prognosis, were assessed. Nuclei polymorphism highly correlated with T stage (p < 0.0001), N stage (p < 0.046), and metastases to the lymph nodes pN (p < 0.004 ). Also, the number of cells in irregular mitosis correlated with T stage (p < 0.004), and highly with pN (p < 0.009). The significance of CDKN2A hypermethylation as a good prognostic factor was also established in the Kaplan-Meir test. The ultrastructural analysis showed that none of the examined tumors had homogenous texture and that resection margin specimens clean in HE stained tissue samples frequently contained single tumor cells or few cells in groups surrounded by connective tissue. This indicates the superiority of electron microscopy over standard histopathological analysis. Thus, a combination of such morphological examination with epigenetic parameters described herein could result in the discovery of promising new prognostic markers of the disease.

Variable course of progression of oral cavity and oropharyngeal carcinoma in young adults

Aim of the study The main aim of this article is the epidemiological analysis of patients treated due to oral and oropharyngeal cancer, with a special interest in the group under the age of 40, evaluation of the differences in the clinical course of the disease as well as assessment of the treatment results, regarding the age of the affected individuals. Material and methods 523 individuals affected by oral and oropharyngeal cancer who were treated in the Otolaryngology and Laryngeal Oncology Department between 2000 and 2008. Precise analysis was performed on 360 out of 523 affected individuals, in whom full clinical status was determined; 13 patients were young adults. The retrospective analysis was created based on case histories, surgical protocols and emergency records. Results In young adults there is a markedly increased risk of organ involvement according to the Mann-Whitney U-test analysis (p = 0.044907). The probability of recurrence in the group of young adults is also much higher. Analysis of Kaplan-Meier test results indicated that the chance for the lack of recurrence within a 6-month period was 85.7%; however, the risk of recurrence increased, and after 12 months was equal to the arithmetic data (50%). Conclusions Among young adults there is an increased risk of local recurrence following 12 months after surgical intervention. According to our observations, despite rapid progression and early recurrence in young adults (4/7) the prognosis for both groups is not statistically different.

Retrospektywna analiza przyczyn niepowodzeń w leczeniu nowotworów jamy ustnej i gardła w Klinice Otolaryngologii i Onkologii Laryngologicznej Uniwersytetu Medycznego w Poznaniu w latach 2000–2005

Summary Aim Retrospective analysis of treatment results in the patients group with oral cancer treated in Department of Otolaryngology and Clinical Oncology K. Marcinkowski University of Medical Sciences in Poznan between the years 2000–2005. The critical view of the cause of the failure. Material The clinical data include: sex, age, stage of the tumor (TNM classification), histological grading, nodes status and 5-years follow up. The parameters were analysed regarding to history of the patients, operative books, outpatients documentation. All parameters were underwent statistical analysis using following tests: Kaplan-Meier, Wilcoxon regarding Gehan and chi2. Data for analysis was classified as „end feature” (death or lack of postoperative control) and „cut feature” (patient still under control). Results Between 2000 and 2005 263 patients were operated due to oral cancer. Only 144 undergo analysis. The rest of 108 patients were calculated as a treatment failure. The middle age was 63 years. There was statistical correlation between tumor stages T (p≤0.00768), pathological N stage (p≤0.07225) and 5 year survival. No correlation was found between age, extracapsular spread of the nodes and histopathological grading G and prognosis. In our group 5-years survival present 32% of the patients. Conclusions The results of 5-years survival in our patients group are still unsatisfactory. The key lies probably on both side. The first the patients visited doctors still too late with significant advancement of the tumor and second we need better education, introduction of new technologies and shorter time for histological diagnosis and waiting list for post operative radiochemioterapy.

9-letnie doświadczenia Kliniki Poznańskiej w chirurgii czynnościowej zatok przynosowych – od mikroskopu operacyjnego do techniki czterech rąk

Summary Surgical procedure is used in chronic rhinosinusitis and aims at restoring proper drainage and ventilation. For this reason the surgery of this region is referred to as functional and in English literature is termed as FESS. It supplanted classical method of Caldwell-Luc or Denker. In the study nine years experience in the Department of Otolaryngology, Head and Surgery University of Poznan in the treatment of chronic sinusitis has been presented. 1463 patients treated with FEES were analyzed. Moreover, the method of selecting patients for the surgery, the diagram showing surgeries performed step by step as well as a list of various surgeries and intraoperative complications were presented. This work is the first part of the study. In the other part the results of the treatment based on the questionnaires sent to patients will be presented.

Frequent methylation of DAB2, a Wnt pathway antagonist, in oral and oropharyngeal squamous cell carcinomas

BACKGROUND Aberrations in Wnt signaling pathway are related to the pathogenesis of head and neck carcinomas and their activation frequently results from epigenetic alterations. This study aimed to assess the frequency of the methylation of DAB2, which acts as a negative regulator of Wnt signaling, and correlate it with clinicopathological features in a group of oral cancer patients. MATERIAL AND METHODS Forty nine patients with primary oral squamous cell carcinoma were enrolled in the study. DNA samples were isolated from surgical sections using phenol-chloroform extraction. Methylation-specific PCR was used to detect gene promoter methylation. RESULTS The analysis of the occurrence of DAB2 promoter methylation in primary oral carcinomas indicated that the gene is methylated in 70% of cases. However, no correlation was found between its methylation and TNM staging or overall survival. CONCLUSIONS Our findings corroborate that DAB2 is a frequent target of epigenetic silencing in oral carcinomas and may be potentially used for tumor detection.

Functional endoscopic sinus surgery (FESS) in unilateral sinus disease

BACKGROUND Unilateral sinus disease (USD) occurs in 23 % of all cases. It is believed that it is mainly associated with cancer development. Retrospective data from large rhinological centers show that the most common USD is chronic rhinosinusitis (CRS), followed by mycosis, inverted papilloma and finally cancer, but only in a small percent of cases. THE AIM OF THE STUDY The analysis of USD in the group of patients who underwent FESS at the secondary referral center. MATERIAL AND METHOD The retrospective study of patients treated for USD in the Department of Otolaryngology in the Provincial Hospital in Poznan between June 2014 and June 2016. The analysis includes age, sex, the localization of lesions, histopathological and microbiological results, an extension of the surgery and treatment results. RESULTS Over the analyzed period of time, 415 FESS for chronic sinusitis were performed. In this group, 83 patients underwent surgery for USD. There were 35 women and 48 men.CRS was found in 48 cases, mycosis in nine cases, 12 patients were operated for non-malignant tumors, such as inverted papilloma (9), osteoma (2) and fibrosis tumor(1); seven patients had a choanal polyp and two of them had a foreign body in maxillary sinus - a tooth root lying loose. Four patients were diagnosed with a hypoplastic maxillary sinus and one patient suffered from frontal sinus pyocele. One side endoscopic opening of all sinuses was performed in the group with CRS, endoscopic medial maxillectomy was conducted in patients with inverted papilloma, and an isolated opening of the affected sinus was performed in the cases with mycosis. CONCLUSIONS USD must be always suspected of malignant degeneration until proven otherwise. Endoscopic sinus surgery with the use of angled scope allows for the removal of even very extensive lesions. In our opinion, the extent of operation is determined by the nature of pathology. While extensive surgery is recommended in patients with inverted papilloma, a limited procedure should be performed in those with isolated mycosis.

Rola karbocysteiny w leczeniu zapaleń zatok przynosowych

Summary Chronic sinusitis is one of the most common presenting complaints of all doctor visits in the United States and Europe, with more than 13% of people affected in any given year. This disease has a wide range of impact on communities. Patients with recurrent or chronic sinusitis report a deteriorative sense of general health and vitality, when compared to general population. In our Department we perform about 600 functional endoscopic sinus surgeries (FESS) per year. Chronic rhinosinusitis represents a spectrum of inflammatory and infectious processes concurrently affecting the nose and paranasal sinuses. Among chronic paranasal sinusitis one must single out paranasal sinusitis with and without polyps. In the paranasal sinusitis patomechanism the blockage of natural ostium plays one of the most important roles. The closure of sinus proper ventilation passages leads to the triggering of many pathological occurrences within mucous membrane of this region. The treatment of paranasal sinusitis is diversified and involves a surgical procedure as well as anti-inflammatory and antiallergic drugs (medications) and mucolytics. Its purpose is to clear the nose through the elimination of bacterial infection, liquidating and removal of lying discharge and the restoration of the proper muco-ciliary transportation, and through this the improvement of local condition and faster recovery. In this work the usage of carboxycysteine to treat paranasal sinus conditions has been presented.

Znaczenie postępowania pooperacyjnego w operacjach czynnościowych nosa i zatok przynosowych

Summary Introduction In this work we wish to emphasize the importance of postoperative care of patients who underwent FESS in ENT Department Medical University in Poznan. We feel that too little attention is still being paid to this treatment as we mostly focus on surgery. Material and methods Our observation are based on nine year experience in functional endoscopic sinus surgery. The analysis covers patients hospitalized in our Department from January 1998 to August 2007. The schedule of postoperative care has been presented. Results 1463 FESS operation were carried out in our Department from January 1998 to August 2007. Similar postoperative procedures were used with each patient. Full recovery of mucous membrane depended on a few parameters such as: if the patient underwent FESS for the first time, what kind of changes were observed: polips or chronic infection, as well as the method of surgery (“aggressive” versus “delicate”). Healing time ranged from 6 week to 3 month. Conclusions The scheme of procedures employed comes useful in postoperative treatment after FESS.