- Email: [email protected]
P1.113. Hypermethylation of genes p14ARF, p16INK4a and MGMT on oral scrapings of patients with oral lichenoid disease
and 1mg of total salivary protein were separated using two-dimensional (2-D) gel electrophoresis over a pH range between 3 and 10 L (18 cm). Spot demarcation and matching was performed through ImageMaster 5.0. Results: A mean of 226 spots were identified in the 3 leukoplakia 2-D gels, and a mean of 262.3 spots in control group. A total of 630 pairs were made among gels from diseased and control classes. It is possible to visualize spots up regulated in leukoplakia 2-D gels (Fig. 1). Statistics and spot identification are under investigation. Discussion: Differences in salivary protein profile in 2-D gel electrophoresis from control and oral leukoplakia subjects can be observed from gel images. Protein identification will be performed through (MALDI-TOF)-TOF mass spectrometry (MS) analysis. Identification of biomarkers could lead to better understanding of prognosis and behaviour of oral leukoplakias. Financial support: CNPq, FAPERJ, Finep, INCA/HCI. doi:10.1016/j.oos.2009.06.396 Further reading 1. van der Waal I. Potentially malignant disorders of the oral and oropharyngeal mucosa; terminology, classification and present concepts of management Oral Oncol. 2009;4–5:317–23. 2. Chen J, He QY, Yuen AP, Chiu JF. Proteomics of buccal squamous cell carcinoma: the involvement of multiple pathways in tumorigenesis. Proteomics 2004;4(8):2465–75. 3. Vitorino R, Lobo MJ, Ferrer-Correira AJ, et al.. Identification of human whole saliva protein components using proteomics. Proteomics 2004;4(4):1109–15. 4. Wong DT. Salivary diagnostics for oral cancer. J Calif Dent Assoc 2006;34(4):303–8.
P1.111. Mutant P53 and possible markers of premalignancy in chronic hyperplastic candidiasis C. McCord, L. Jackson-Boeters, T.D. Daley, M.R. Darling * University of Western Ontario, Canada Introduction: Chronic hyperplastic candidiasis (CHC) is commonly diagnosed in the oral cavity and has previously been associated with cancerous change. McCullough et al. (2002) have shown that there is an association between oral yeast carriage and epithelial dysplasia. P53 is a tumor suppressor gene and an apoptosis regulator, which interacts with many other genes, protecting the cell against DNA damage. The objective of this study was to examine markers which have been associated with malignancy, in CHC. Methods: Immunohistochemical methods were used to examine the expression of p53 wild-type (WT) and mutant (M), MDM2, p21, metallothionein (MT) and proliferating cell nuclear antigen (PCNA) in 41 CHC lesions, 20 candida-infected (CIC) and 11 non-infected control (NIC) tissues. Appropriate positive and negative controls were used. A well-documented scoring system, for intensity and proportion of epithelial cells stained, was used. TUNEL was used to examine apoptosis which was correlated with p53 and metallothionein expression. These markers were measured in sites/lesions of CHC which did not show any epithelial dysplasia, or histological signs of malignancy. Results: Mutated p53 scores were higher in CHC than in controls. Levels of MDM2 were not elevated. P21was increased in parabasal epithelial cells. CHC lesions showed a similar basal/parabasal MT staining pattern seen in normal squamous epithelium. PCNA ap-
159
peared increased in lesions of CHC and CIC, indicating proliferating epithelial cells. Apoptosis was not increased in lesions of CHC. Conclusions: The presence of mutated p53 in oral chronic hyperplastic candidiasis suggests that there may be an increased potential for malignant change to occur in the epithelium, above that of normal tissues. Further, these results suggest a non-active wild-type 53 (perhaps due to activity of MDM2 in at least some cells) and therefore increased potential for DNA damage. Further investigation is required as well as clinical follow-up studies. doi:10.1016/j.oos.2009.06.397
P1.112. Evaluation of a fast-track service for the provision of radiologically inserted gastrostomy (RIG) tubes in head and neck cancer patients M.J. Monteiro *, I. Francis, G. Burkill, J. Herold, K. Altman, S. Koshal Brighton and Sussex University Hospitals NHS Trust, United Kingdom Introduction: Gastrostomy feeding is an important method of providing enteral nutrition in head and neck cancer patients. At the Royal Sussex County Hospital, a fast-track service has been established for the provision of RIGs in head and neck cancer patients, in order that cancer treatment is not delayed. The purpose of this study was to evaluate this service and assess the complications of RIGs. Method: We retrospectively analysed the notes of all patients with head and neck cancer who had undergone RIG insertion at the Royal Sussex County Hospital between 2004 and 2008 to record minor and major complications, and time from patient referral to RIG insertion. The RIG procedure was performed by four operators using a standardised technique. Results: 110 patients underwent RIG insertion at the Royal Sussex County Hospital over a 5 year period. All patients had squamous cell carcinoma of the upper aero-digestive tract. RIG insertion was performed following discussion by the MDT (multidisciplinary team) and prior to surgical resection and chemoradiotherapy. This was coordinated by the nutritional support and nursing teams to ensure the patients were knowledgeable in relation to both the procedure and usage of the RIG. One (0.009%) major (requiring laparotomy) and 6 (5.4%) minor (localised peritonism, haematoma formation and localised sepsis) complications were reported. Discussion: Our study demonstrates that a fast-track service for RIG insertion, guided by a multi-disciplinary meeting, can produce safe and expeditious results with improved nutritional status for patients. This model could be adopted by other units. doi:10.1016/j.oos.2009.06.398
P1.113. Hypermethylation of genes p14ARF, p16INK4a and MGMT on oral scrapings of patients with oral lichenoid disease A. Acha-Sagredo *, X. Marichalar, N. Rey, M.A. Martinez de Pancorbo, M.A. Echebarria, J.M. Aguirre University of the Basque Country EHU, Spain Introduction: Aberrant promoter hypermethylation seems to be an essential step for the development of multiple genetic events which lead to tumour progression. In oral carcinogenesis, inactivation due to methylation has been associated with the development
Poster Abstracts Oral AbstractsPoster ListOrals ListPan. Disc. & Symp. Abs.Keynote Abs.Keynote Bios.ProgramIAOOWelcomeCommittee Listings
Poster session I / Oral Oncology Supplement 3 (2009) 123–161
Poster Abstracts Oral AbstractsPoster ListOrals ListPan. Disc. & Symp. Abs.Keynote Abs.Keynote Bios.ProgramIAOOWelcomeCommittee Listings
160
Poster session I / Oral Oncology Supplement 3 (2009) 123–161
and progression of this pathology. However, little is known about this genetic alteration in a common premalignant condition such as oral lichenoid disease (OLD). OLD represents a group of chronic processes with an immunological basis which malignant potential is controversial. Objective: The aim of our study was to assess the promoter methylation status of genes p14ARF, p16INK4a and MGMT in oral scrapings of patients with oral lichenoid disease. Methods: Sixty-eight cytological samples of patients with OLD (57.76 years mean age, 47 women and 21 men) were studied. DNA was extracted from the oral scraping and the methylation status of the promoter regions of p14ARF, p16INK4a and MGMT were analyzed by quantitative methylation specific PCR (QMSP). Results: Tumour suppressor gene p14ARF showed promoter hypermethylation in one case, and DNA repair gene MGMT showed promoter hypermethylation in two cases (one case developed squamous cell carcinoma). No promoter hypermethylation was found in p16INK4a. Discussion: Promoter hypermethylation of genes p14ARF, p16 INK4a and MGMT does not seem to be common in patients with oral lichenoid disease. Grant supports: Carlos III Health Institute – FIS (PI051400) and Department of Education, Universities and Research, Government of the Basque Country (IT-192-07). doi:10.1016/j.oos.2009.06.399
P1.114. The development of in vitro models of squamous cell carcinoma of the oral mucosa – From mild dysplasia to an expanding carcinoma H.E. Colley a,b, S. MacNeil a, A.V. Jones b, M.H. Thornhill b,*, C. Murdoch b a
Department of Engineering Materials, University of Sheffield, United Kingdom b School of Clinical Dentistry, University of Sheffield, United Kingdom Worldwide, there are 450,000 new cases of oral squamous cell carcinoma (OSCC) every year. OSCC evolves from normal epithelium through mild, moderate and severe dysplasia to carcinoma in situ before invading the basement membrane and connective tissue to form a carcinoma. The aim of this study was to establish novel in vitro models of OSCC that closely replicate the in vivo situation at different stages of disease progression. OSCC cell lines (FaDu and Cal27) were seeded into tissue engineered human oral mucosa (TEOM) both as multi-cellular tumour spheroids (MCTS) and as cell suspensions. These models were cultured for up to 21 days and compared histologically with naturally occurring early tumours from patient biopsies. The OSCC FaDu cell line formed MCTS within 18 h. The MCTS had predictable growth in culture with a proliferating outer layer, hypoxic inner rim and necrotic centre that resembled an expanding tumour island. When seeded within the TEOM, FaDu MCTS produced a histological picture mimicking carcinoma in situ with highly disorganised epithelial architecture and severe cellular atypia juxtaposed to normal epithelium. When added as a suspension, Cal 27 tumour cells traversed the basement membrane and invaded the connective tissue underlying a severely dysplastic epithelium to reproduce the features of early invasive OSCC. These results demonstrate that the addition of OSCC to TEOM reproduces the in vivo appearance of early stage disease. Whereas, cells cultured as MCTS replicate the expanding tumour islands seen in more a more advanced disease state. Using different techniques, it
is possible to model OSCC at different stages of progression, from early lesions through to a developing carcinoma. Such models could facilitate study of the disease processes involved in malignant transformation, early invasion and tumour growth as well as in vitro testing of new treatments, diagnostic tests and drug delivery systems for OSCC. doi:10.1016/j.oos.2009.06.400
P1.115. Expression of epidermal growth factor receptor (EGFR) in oral lichen planus D.A. Cortés *, M.A. Gainza, M.J. Rodriguez-Tojo, R. Martinez-Conde, J.M. Aguirre University of the Basque Country EHU, Spain Introduction: The epidermal growth factor receptor (EGFR) is an important pathway in cancer development. Protein overexpression might be responsible for activation of the EGFR pathway in head and neck carcinogenesis. Oral lichen planus (OLP) is a chronic inflammatory premalignant disease, the risk for transformation into squamous cell carcinoma of the head and neck (SCCHN) is a matter of controversy, related with different clinicopathological subtypes. There is not reliable biomarker to identify those OLP lesions with increased risk malignant potential. Purpose: To determine the immunoexpression of EGFR in biopsies of OLP. Patients, materials and methods: We studied 44 cases of OLP, 23 cases (52.3%) histopathologically classified as typical (HPT) and 21 (47.7%) as compatible (HPC). Standard immunostaining method was performed with monoclonal antibody against EGFR protein (clone 31G7 ZymedÒ). The main clinicopathological data was collected based on a previous protocol. Statistical software SPSS 15.0. was used for a comparative and descriptive analysis. Results: Expression of EGFR was detected in the epithelium of 100% (44) of the samples, 9 (20.5%) with low expression and 35 (79.5%) high expression. There were significant differences related to cytoplasmatic expression between HPT and HPC groups. The lesions with parakeratosis show higher expression. Conclusion: There are differences in EGFR expression in OLP lesions related to its histopathological aspects and can be used to differentiate the clinicopathological subtypes. Grant supports: Carlos III Health Institute – FIS (PI051400) and Department of Education, Universities and Research, Government of the Basque Country (IT-192-07). doi:10.1016/j.oos.2009.06.401
P1.116. Evaluation of diagnostic aids for the detection of oral potentially malignant disorders K.H. Awan *, S. Warnakulasuriya King’s College London, United Kingdom Objectives: Early detection of oral cancer is crucial in improving survival rate. A variety of new and emerging diagnostic aids and adjunctive techniques are currently available to potentially assist in the detection of oral potentially malignant disorders (OPMDs). The accuracy of three adjunctive tests namely Toluidine blue staining, Autofluorescence, and Chemiluminescence was evaluated in relation to conventional oral examination and surgical biopsy (gold standard).
P1.111. Mutant P53 and possible markers of premalignancy in chronic hyperplastic candidiasis C. McCord, L. Jackson-Boeters, T.D. Daley, M.R. Darling * University of Western Ontario, Canada Introduction: Chronic hyperplastic candidiasis (CHC) is commonly diagnosed in the oral cavity and has previously been associated with cancerous change. McCullough et al. (2002) have shown that there is an association between oral yeast carriage and epithelial dysplasia. P53 is a tumor suppressor gene and an apoptosis regulator, which interacts with many other genes, protecting the cell against DNA damage. The objective of this study was to examine markers which have been associated with malignancy, in CHC. Methods: Immunohistochemical methods were used to examine the expression of p53 wild-type (WT) and mutant (M), MDM2, p21, metallothionein (MT) and proliferating cell nuclear antigen (PCNA) in 41 CHC lesions, 20 candida-infected (CIC) and 11 non-infected control (NIC) tissues. Appropriate positive and negative controls were used. A well-documented scoring system, for intensity and proportion of epithelial cells stained, was used. TUNEL was used to examine apoptosis which was correlated with p53 and metallothionein expression. These markers were measured in sites/lesions of CHC which did not show any epithelial dysplasia, or histological signs of malignancy. Results: Mutated p53 scores were higher in CHC than in controls. Levels of MDM2 were not elevated. P21was increased in parabasal epithelial cells. CHC lesions showed a similar basal/parabasal MT staining pattern seen in normal squamous epithelium. PCNA ap-
159
peared increased in lesions of CHC and CIC, indicating proliferating epithelial cells. Apoptosis was not increased in lesions of CHC. Conclusions: The presence of mutated p53 in oral chronic hyperplastic candidiasis suggests that there may be an increased potential for malignant change to occur in the epithelium, above that of normal tissues. Further, these results suggest a non-active wild-type 53 (perhaps due to activity of MDM2 in at least some cells) and therefore increased potential for DNA damage. Further investigation is required as well as clinical follow-up studies. doi:10.1016/j.oos.2009.06.397
P1.112. Evaluation of a fast-track service for the provision of radiologically inserted gastrostomy (RIG) tubes in head and neck cancer patients M.J. Monteiro *, I. Francis, G. Burkill, J. Herold, K. Altman, S. Koshal Brighton and Sussex University Hospitals NHS Trust, United Kingdom Introduction: Gastrostomy feeding is an important method of providing enteral nutrition in head and neck cancer patients. At the Royal Sussex County Hospital, a fast-track service has been established for the provision of RIGs in head and neck cancer patients, in order that cancer treatment is not delayed. The purpose of this study was to evaluate this service and assess the complications of RIGs. Method: We retrospectively analysed the notes of all patients with head and neck cancer who had undergone RIG insertion at the Royal Sussex County Hospital between 2004 and 2008 to record minor and major complications, and time from patient referral to RIG insertion. The RIG procedure was performed by four operators using a standardised technique. Results: 110 patients underwent RIG insertion at the Royal Sussex County Hospital over a 5 year period. All patients had squamous cell carcinoma of the upper aero-digestive tract. RIG insertion was performed following discussion by the MDT (multidisciplinary team) and prior to surgical resection and chemoradiotherapy. This was coordinated by the nutritional support and nursing teams to ensure the patients were knowledgeable in relation to both the procedure and usage of the RIG. One (0.009%) major (requiring laparotomy) and 6 (5.4%) minor (localised peritonism, haematoma formation and localised sepsis) complications were reported. Discussion: Our study demonstrates that a fast-track service for RIG insertion, guided by a multi-disciplinary meeting, can produce safe and expeditious results with improved nutritional status for patients. This model could be adopted by other units. doi:10.1016/j.oos.2009.06.398
P1.113. Hypermethylation of genes p14ARF, p16INK4a and MGMT on oral scrapings of patients with oral lichenoid disease A. Acha-Sagredo *, X. Marichalar, N. Rey, M.A. Martinez de Pancorbo, M.A. Echebarria, J.M. Aguirre University of the Basque Country EHU, Spain Introduction: Aberrant promoter hypermethylation seems to be an essential step for the development of multiple genetic events which lead to tumour progression. In oral carcinogenesis, inactivation due to methylation has been associated with the development
Poster Abstracts Oral AbstractsPoster ListOrals ListPan. Disc. & Symp. Abs.Keynote Abs.Keynote Bios.ProgramIAOOWelcomeCommittee Listings
Poster session I / Oral Oncology Supplement 3 (2009) 123–161
Poster Abstracts Oral AbstractsPoster ListOrals ListPan. Disc. & Symp. Abs.Keynote Abs.Keynote Bios.ProgramIAOOWelcomeCommittee Listings
160
Poster session I / Oral Oncology Supplement 3 (2009) 123–161
and progression of this pathology. However, little is known about this genetic alteration in a common premalignant condition such as oral lichenoid disease (OLD). OLD represents a group of chronic processes with an immunological basis which malignant potential is controversial. Objective: The aim of our study was to assess the promoter methylation status of genes p14ARF, p16INK4a and MGMT in oral scrapings of patients with oral lichenoid disease. Methods: Sixty-eight cytological samples of patients with OLD (57.76 years mean age, 47 women and 21 men) were studied. DNA was extracted from the oral scraping and the methylation status of the promoter regions of p14ARF, p16INK4a and MGMT were analyzed by quantitative methylation specific PCR (QMSP). Results: Tumour suppressor gene p14ARF showed promoter hypermethylation in one case, and DNA repair gene MGMT showed promoter hypermethylation in two cases (one case developed squamous cell carcinoma). No promoter hypermethylation was found in p16INK4a. Discussion: Promoter hypermethylation of genes p14ARF, p16 INK4a and MGMT does not seem to be common in patients with oral lichenoid disease. Grant supports: Carlos III Health Institute – FIS (PI051400) and Department of Education, Universities and Research, Government of the Basque Country (IT-192-07). doi:10.1016/j.oos.2009.06.399
P1.114. The development of in vitro models of squamous cell carcinoma of the oral mucosa – From mild dysplasia to an expanding carcinoma H.E. Colley a,b, S. MacNeil a, A.V. Jones b, M.H. Thornhill b,*, C. Murdoch b a
Department of Engineering Materials, University of Sheffield, United Kingdom b School of Clinical Dentistry, University of Sheffield, United Kingdom Worldwide, there are 450,000 new cases of oral squamous cell carcinoma (OSCC) every year. OSCC evolves from normal epithelium through mild, moderate and severe dysplasia to carcinoma in situ before invading the basement membrane and connective tissue to form a carcinoma. The aim of this study was to establish novel in vitro models of OSCC that closely replicate the in vivo situation at different stages of disease progression. OSCC cell lines (FaDu and Cal27) were seeded into tissue engineered human oral mucosa (TEOM) both as multi-cellular tumour spheroids (MCTS) and as cell suspensions. These models were cultured for up to 21 days and compared histologically with naturally occurring early tumours from patient biopsies. The OSCC FaDu cell line formed MCTS within 18 h. The MCTS had predictable growth in culture with a proliferating outer layer, hypoxic inner rim and necrotic centre that resembled an expanding tumour island. When seeded within the TEOM, FaDu MCTS produced a histological picture mimicking carcinoma in situ with highly disorganised epithelial architecture and severe cellular atypia juxtaposed to normal epithelium. When added as a suspension, Cal 27 tumour cells traversed the basement membrane and invaded the connective tissue underlying a severely dysplastic epithelium to reproduce the features of early invasive OSCC. These results demonstrate that the addition of OSCC to TEOM reproduces the in vivo appearance of early stage disease. Whereas, cells cultured as MCTS replicate the expanding tumour islands seen in more a more advanced disease state. Using different techniques, it
is possible to model OSCC at different stages of progression, from early lesions through to a developing carcinoma. Such models could facilitate study of the disease processes involved in malignant transformation, early invasion and tumour growth as well as in vitro testing of new treatments, diagnostic tests and drug delivery systems for OSCC. doi:10.1016/j.oos.2009.06.400
P1.115. Expression of epidermal growth factor receptor (EGFR) in oral lichen planus D.A. Cortés *, M.A. Gainza, M.J. Rodriguez-Tojo, R. Martinez-Conde, J.M. Aguirre University of the Basque Country EHU, Spain Introduction: The epidermal growth factor receptor (EGFR) is an important pathway in cancer development. Protein overexpression might be responsible for activation of the EGFR pathway in head and neck carcinogenesis. Oral lichen planus (OLP) is a chronic inflammatory premalignant disease, the risk for transformation into squamous cell carcinoma of the head and neck (SCCHN) is a matter of controversy, related with different clinicopathological subtypes. There is not reliable biomarker to identify those OLP lesions with increased risk malignant potential. Purpose: To determine the immunoexpression of EGFR in biopsies of OLP. Patients, materials and methods: We studied 44 cases of OLP, 23 cases (52.3%) histopathologically classified as typical (HPT) and 21 (47.7%) as compatible (HPC). Standard immunostaining method was performed with monoclonal antibody against EGFR protein (clone 31G7 ZymedÒ). The main clinicopathological data was collected based on a previous protocol. Statistical software SPSS 15.0. was used for a comparative and descriptive analysis. Results: Expression of EGFR was detected in the epithelium of 100% (44) of the samples, 9 (20.5%) with low expression and 35 (79.5%) high expression. There were significant differences related to cytoplasmatic expression between HPT and HPC groups. The lesions with parakeratosis show higher expression. Conclusion: There are differences in EGFR expression in OLP lesions related to its histopathological aspects and can be used to differentiate the clinicopathological subtypes. Grant supports: Carlos III Health Institute – FIS (PI051400) and Department of Education, Universities and Research, Government of the Basque Country (IT-192-07). doi:10.1016/j.oos.2009.06.401
P1.116. Evaluation of diagnostic aids for the detection of oral potentially malignant disorders K.H. Awan *, S. Warnakulasuriya King’s College London, United Kingdom Objectives: Early detection of oral cancer is crucial in improving survival rate. A variety of new and emerging diagnostic aids and adjunctive techniques are currently available to potentially assist in the detection of oral potentially malignant disorders (OPMDs). The accuracy of three adjunctive tests namely Toluidine blue staining, Autofluorescence, and Chemiluminescence was evaluated in relation to conventional oral examination and surgical biopsy (gold standard).