O111 Towards a rapid assay for detection of residual cancer

94 Keywords: quality of life, University of Washington quality of life scale, validation, function O108 Predictors of quality of life after treatment for oral cancer T. Gurney *, D. Eisele, L. Orloff, S. Wang. University of California, United States Introduction: The purpose of this study was to analyze the various demographic, treatment, and outcome variables that may affect quality of life following oral cancer treatment. A cross-sectional study of patients previously treated for oral cavity and oropharyngeal cancers was performed. Methods: All patients previously treated for oral cavity and oropharynx carcinoma seen at a tertiary head and neck surgery clinic over a four month period were requested to participate in this study. Participants completed the University of Michigan Head & Neck Specific Quality of Life questionnaire, which is a validated survey instrument that analyzes the effects of cancer treatment on 4 domains, including eating, speech, emotion and pain. Survey results were analyzed with respect to various patient demographic, treatment, and outcome parameters to determine variables that predicted perceived quality of life differences. Results: Eighty-seven patients eligible for the study completed questionnaires. The majority of study subjects were male (62%). Analysis of the various treatment modalities received by patients in this study revealed that a history of surgery did not significantly impact perceived quality of life in any domain. On the other hand, a history of radiation treatment or surgery with radiation treatment predicted significantly poorer quality of life scores in the domain of eating and the domains of eating and speech, respectively (p < 0.05). Other predictors of poorer quality of life scores included advanced stage (eating domain), gastrostomy-tube dependence (eating, speech, and emotion domains), surgical complications (eating and emotion domains), and cancer recurrence (eating, emotion, and speech domains). Discussion: Quality of life among oral cancer patients is negatively affected by advanced stage, radiation treatment, surgery with radiation treatment, complications after surgery, gastrostomy-tube dependence, and cancer recurrence. Knowledge of negative predictors of quality of life can allow physicians to provide better supportive care where appropriate for oral cancer patients. Keywords: oral cancer, quality of life, radiation therapy O109 Defect characteristics and type of reconstruction as predictors of speech and swallowing following surgery for oral and oropharyngeal cancer A. Schache *, N. Kalavrezos, J. Roe, L. Newman, C. Hopper. University College London Hospitals, United Kingdom Speech and swallowing function in postsurgical head and neck cancer patients may be affected by the degree of resection and the mode of reconstruction. Articulation and the oral stage of the swallow are generally more severely impaired, although the pharyngeal stage of the swallow may be affected if resection includes the tongue base. This study involves subjective and video swallow assessment in a sample of 40 patients who received surgery as primary treatment for intraoral cancer at UCLH. Our aim was to correlate the functional outcome to the site and the extent of the resection of the oral cavity, to the type of reconstruction and to postoperative radiotherapy. There were four categories of defect depending on the site of resection: 1. lateral-anterior defects including the lateral floor of mouth, buccal vestibule and/or the mandible

Oral and Symposium abstracts, Saturday 19 May 2. central-anterior defects including the anterior floor of mouth, labial vestibule and/or the mandible (intercaninal segment) 3. posterior-lateral defects including the retromolar area, lateral pharyngeal wall and/or the posterior mandible 4. central defects including the tongue and the floor of mouth Most of the defects were reconstructed with a free vascularised flap whereas in few cases a pedicled regional flap was used. A combination of free and pedicled flap was used in one patient. All patients included had a subjective speech assessment in the immediate postoperative period whereas a video swallow was performed in most cases as a baseline assessment prior to the commencement of radiotherapy. The rest of the patients had a video swallow in the post-radiotherapy set up. The results of the subjective and video swallow assessment are correlated to the site, the extent of the resection and the type of reconstruction. The impact of postoperative radiotherapy on oral functions is also discussed. Keywords: speech, swallow, oral cancer, reconstruction Basic science III O110 Increased CK19 expression correlated with pathological differentiation grade and prognosis in oral squamous cell carcinoma patients L. Zhong *, Z. Zhang, C. Zhang, W. Chen. Shanghai Jiao Tong University, China Objective: To investigate cytokeratin 19 (CK19) expression in oral squamous cell carcinoma (OSCC) tissue as well as its correlation with pathological differentiation grade and prognosis. Materials and Methods: Cancerous and distant tissues from 33 primary OSCC patients were collected during surgical operation. Each sample was divided into two parts, one for CK19 protein detection using immunohistochemistry, and the other for CK19 mRNA level detection of relative quantification using fluorescent real-time RT-PCR. Results: CK19 positive rate in cancerous tissue was 90.9% (30/33) detected using immunohistochemistry, which was significantly higher than that in distant tissue (15.2%, 5/33) by Chisquare test (P < 0.001). CK19 positive score in cancerous tissue was also significantly higher than that in the corresponding distant tissue by Paired samples t-test (P < 0.001). Patients with positive CK19 expression in distant tissue had higher tumor recurrence rate (P = 0.008) and lower survival rate (P = 0.015) than those with negative CK19 expression in distant tissue. Using fluorescent real-time RT-PCR, CK19 mRNA level in cancerous tissue was 2.21 folds higher than that in distant tissue (P = 0.020). CK19 expression correlated significantly with pathological differentiation grade, which was detected not only using immunohistochemistry (P = 0.002) but also using fluorescent real-time RT-PCR (P = 0.025), poorer differentiated indicating higher CK19 positive score and CK19 mRNA level. Conclusions: Increased CK19 protein expression and gene transcription in OSCC tissue correlate significantly with pathological differentiation grade. Positive CK19 expression in distant tissue suggests a higher tumor recurrence rate and a lower survival rate. Keywords: cytokeratin 19, oral squamous cell carcinoma, pathological differentiation grade, prognosis O111 Towards a rapid assay for detection of residual cancer X. Huang *. King’s College, United Kingdom Introduction: Molecular detection of residual cancer may improve prediction of cases likely to develop a recurrence after surgery. The p53 phage plaque assay provides the current gold standard method for detection of residual cancer. However,

this multi-step approach is cumbersome, time consuming and more rapid approaches are needed if this technology is to be translated from the laboratory to the clinic. The gap ligase chain reaction (gap-LCR, Harden SV, Clinical Cancer Res 2004:10, 2379) may provide a suitable alternative strategy for molecular analysis of ‘molecular’ margins. Methods: Cases (n = 15) were analysed with the p53 phage plaque assay as described by Partridge et al (Clinical Cancer Res 2000:6, 2718) and the gap-LCR. A similar number of cases were analysed with gap-LCR only. Refinements were made to the protocol for the gap-LCR (incorporation of primers with mismatches, additional wild type primers as well as varying the concentration of template and reaction conditions) to increase the ability of the assay to detect low levels of mutant p53. Results: Eleven cases were scored as tumour-positive positive with the plaque assay and 6 with gap-LCR; 4 cases were negative with both tests. One hundred and eleven surgical margins were analysed. The concordance between the results obtained with both assays was poor. The sensitivity of a tumourpositive test, as obtained with the phage plaque assay for predicting local recurrence, was 85.7% and the specificity 37.5%. The gap-LCR had a lower sensitivity (57%) but a higher specificity (75%). The ability of the gap-LCR test to detect low levels of some mutant p53 sequences could be improved with the modifications outlined above, but overall this test did not provide a satisfactory replacement for the phage plaque assay. Conclusions: The gap-LCR assay does not have sufficient sensitivity to replace the phage plaque assay for a range of p53 mutations and cannot be recommended as an alternative at this time. Keywords: p53, residual cancer, molecular diagnostics O112 Behavior of the most invasive oral squamous cell carcinoma (Gr.4D) E. Yamamoto *, S. Kawashiri, K. Kato, T. Ohara, H. Kitahara. Kanazawa University, Japan Introduction: The histologic grading of mode(or pattern) of invasion in squamous cell carcinoma of head and neck was first proposed by Jakobsson in 1973. Grade 4 of mode of invasion, i.e., diffuse invasion was subclassified by us into Grs.4C(Cordlike type) and 4D(Diffuse, widespread type) for oral squamous cell carcinoma (OSCC) in 1983 (Cancer 51:2175 80), with Gr.4D being the poorest prognosis of 29% survial rate (overall:73%). The purpose of this study since 1988 is to clarify the behavior of Gr.4D cells in tumor tissue, in vitro and in vivo. Materials and Methods: (1) Initial biopsy specimens from the patients with OSCC reffered to our clinic were prepared as immunostained sections for microscopic observations. (2) Five cell lines OSC20 (Gr.3), OSC19 (Gr.4C), Hoc313, TSU and OLC1(Grs.4D) were used in vitro invasion assay and (3) in vivo invasion and metastasis model. Results: (1) In immunostained sections from human materials, Gr.4D cases showed almost all loss of basement membrane and massive tumor desmoplasia much more than the other grades. (2) HOC313(Gr.4D) cells showed the greatest motility in in-vitro cell motility assay and the diffusest invasion similar to its origin in cell culture on collagen gel. (3) OSC20 (Gr.3), OSC19 (Gr.4C) and OLC1 (Gr.4D) showed invasion and metastasis similar to their origin in orthotopic implantation model by using nude mice. Discussion and Conclusion: The strongest invasion of Gr.4D cells was shown by comparing with the other grades in human materials, in vitro and in vivo experiments. From these results, Gr.4D type invasion appears to be independent type, while there are intermediate types from Grs. 1 to 2, 2 to 3 and 3 to 4C. The incidence of Gr.4D is less than 10% in all patients. But, it is challenging to overcome the highest grade malignancy. In recent years, excisional biopsy for T1N0 to early T2N0 cases

95 is tried to prevent cancer cells dissemination and good results are obtained. Keywords: oral squamous cell carcinoma, mode of invasion, pathology O113 Head and neck squamous cell carcinoma in non-smoking and non-drinking patients with multiple tumors: etiologic significance of p53 overexpression and ki-67 suprabasal expression in non tumorous epithelium F. Farshadpour *, G.J. Hordijk, R. Koole, P. Slootweg. University Medical Center Utrecht, Netherlands Objectives: Non-smoking and non-drinking (NSND) patients with head and neck squamous cell carcinoma (HNSCC) have different clinical characteristics than their smoking and drinking (SD) counterparts. They are predominantly older female patients with oral cavity tumors, however both groups show the same high percentage of second primary tumors. We studied the mucosal expression of tumor suppressor gene p53 and proliferation marker Ki-67 in NSND patients with HNSCC to study possible clonal origin of multiple tumors and gain an understanding of their carcinogenesis. Method: Six groups were selected: NSND patients with multiple HNSCCs (A) and single HNSCC (B), SD patients with multiple HNSCCs (C) and single HNSCC (D), individuals without tumor who smoked tobacco and drank alcohol (E) and those who did not (F). For groups A, B, C and D, we analyzed the overexpression of p53 and suprabasal expression of Ki-67 in non-malignant tumor-adjacent mucosa. The same analysis was performed for groups E and F. The occurrence of these markers in all groups were compared to each other. Results: The occurrence of overexpression of p53 and suprabasal expression of Ki-67 was significantly higher in users of tobacco and alcohol than in non-users. These biomarker expressions were not significantly affected by the number of tumors present. Conclusion: This seems to confirm the relationship between tobacco and alcohol use and the expression of p53 and Ki67. However the significance of these biomarkers in terms of multiple HNSCC development is questionable. Further study of other biomarkers or ploidy analysis should be performed to understand the carcinogenesis of single and multiple HNSCC in NSND patients. Keywords: head and neck squamous cell carcinoma, tobacco, alcohol, carcinogenesis O114 Promoter hypermethylation quantification in oral dysplasia and cancer B. Schmidt *, C. Viet. University of California, United States Background: Promoter hypermethylation, a major mechanism in silencing cancer-associated genes, is potentially an early diagnostic marker for oral cancer. Saliva is an ideal diagnostic fluid because of ease of collection. However, to date there have been no studies quantifying promoter hypermethylation in the saliva of oral dysplasia and cancer patients. Objective: To quantitatively analyze promoter hypermethylation for five genes ( p15, p16, MGMT, E-cadherin, and APC) using tissue and saliva from 3 groups of patients: normal, dysplasia, and cancer. To determine the correlation between saliva and tissue promoter hypermethylation. Methods: Saliva was collected from 11 oral cancer, 3 oral dysplasia and 5 normal subjects. Methylight, a fluorescence based real-time PCR technique, was used to quantify DNA methylation at the locus of interest in the samples. The percentage of fully methylated molecules at the locus of interest, indicated as PMR (percentage of methylated reference), was calculated for each sample and compared to

Oral abstracts

Oral and Symposium abstracts, Saturday 19 May