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P3.36. Overexpression of NRG1 correlating to cisplatin-resistance in oral squamous cell carcinoma
Poster Abstracts Oral AbstractsPoster ListOrals ListPan. Disc. & Symp. Abs.Keynote Abs.Keynote Bios.ProgramIAOOWelcomeCommittee Listings
212
Poster session III et al. / Oral Oncology Supplement 3 (2009) 201–236
doi:10.1016/j.oos.2009.06.560
P3.35. The expression of vascular endothelial growth factor A (VEGFA) and C (VEGFC) and their receptors in head and neck squamous cell carcinoma M. Macluskey a,*, S. Jones a, G. Stenhouse b, P. McLoughlin b, I. Ellis a, H. Alkhadar a a b
University of Dundee, United Kingdom Ninewells Hospital, United Kingdom
Introduction: Angiogenesis is a crucial for tumour growth, progression, and metastasis. Whilst vascular endothelial growth factor A (VEGFA) is an important angiogenic factor, vascular endothelial growth factor C (VEGFC) may stimulate the lymphatic system and facilitate lymphatic metastasis. The aim of this study was to investigate vascular endothelial growth factor A and C (VEGFA and VEGFC) and their receptors VEGFR1 and VEGFR3 in head and neck squamous cell carcinomas (HNSCC) and in nodal metastasis. Methods: Immunohistochemistry was used to identifying blood and lymphatic vessels, VEGF-A, VEGF-C and their receptors. The cohort of patients studied consisted of 22 patients with dysplastic lesions and 64 patients with cancer. Pre-treatment serum levels of VEGFA; VEGFC; VEGFR1 and VEGFR3 were determined in 63 of these HNSCC, 8 dysplasias, and in 29 normal controls using commercially available immunoassays. Results: The blood and lymphatic vessel density, VEGFA, VEGFC, VEGFR1 and VEGFR3 expression was significantly higher in HNSCC than dysplasias and significantly higher in positive nodes compared with negative nodes (p < 0.05). The mean serum levels of VEGFA and VEGFC levels were higher in the cancer group compared with both the normal and dysplastic group (p < 0.05). No association was found between any of the serum markers and nodal metastasis. High expression of VEGFA and VEGFC both within the tissue and the serum was associated with poorer survival (p < 0.05). Discussion: Disease progression from dysplasia to cancer and metastasis is accompanied by increases in both blood and lymphatic vessel density which may be facilitated not only by an up regulation of VEGFA and VEGFC but also their receptors. Systemic expression of VEGFA and VEGFC did not correlate with local expression and did not indicate metastatic disease. High expression of VEGFA and VEGFC may indicate patients with aggressive disease who may benefit from more aggressive management and may provide and chemotherapeutic target. doi:10.1016/j.oos.2009.06.561
P3.36. Overexpression of NRG1 correlating to cisplatin-resistance in oral squamous cell carcinoma S. Keiji a,*, U. Katsuhiro a, F. Kazuaki a, S. Kengo b, O. Kanae c, O. Katsunori a a
Department of Clinical Molecular Biology, Graduate School of Medicine, Chiba University, Chiba, Japan b Department of Molecular Virology, Graduate School of Medicine, Chiba University, Chiba, Japan c Division of Dentistry and Oral and Maxillofacial Surgery, Chiba University Hospital, Chiba, Japan Introduction: Resistance to cisplatin remains a major obstacle to successful treatment of oral squamous cell carcinoma (OSCC). To investigate the molecular mechanism of this resistance, we compared the gene expression profiles between the cisplatin-sensitive
SCC cell lines (Sa-3, H-1, and KB) and the cisplatin-resistant cell lines established from them (Sa-3R, H-1R, and KB-R). Methods: We analyzed the gene expression profiles of cisplatinsensitive and -resistant cell lines using the Affymetrix U133 Plus 2.0 microarray. The identified genes were used for Ingenuity Pathway Analysis Tool to identify important functional networks to cisplatin resistance. A candidate gene was selected from those genes, and then, mRNA expression of the gene was evaluated with real-time quantitative reverse transcriptase-polymerase chain reaction (qRTPCR) in both cisplatin-sensitive and -resistant cell lines. Protein expression was also examined by Western blot analysis and immunohistochemistry. In addition, to confirm the function, the gene expression was down-regulated by the small interfering RNA (siRNA) method in the cisplatin-resistant cell line. Results: Microarray analysis revealed 199 of differentially expressed genes. Network analysis was performed with these genes, and further 51 genes were shown to be crucial for the cisplatin-resistant mechanism. 51 of these genes were mapped to genetic networks and we validated the top-10 up-regulated genes by qRTPCR. NRG1 showed excellent concordance with the microarray data. Thus, we selected NRG1 for investigation. In 48 patients with OSCCs, positive immunohistochemical staining for NRG1 correlated with chemoresistance to cisplatin-based combination chemotherapy (P < 0.01). Furthermore, siRNA-directed suppression of NRG1 expression resulted in enhanced susceptibility to cisplatin. Discussion: These results suggested that overexpression of NRG1 is closely related to cisplatin-resistant mechanism in OSCC cells, and that NRG1 may be an effective chemotherapeutic target for this disease. doi:10.1016/j.oos.2009.06.562
P3.37. Loss of 3p26.3 detected by array-based comparative genomic hybridization as a predictive marker of poor prognosis in patients with oral squamous cell carcinomas K. Uchida a,*, K. Harada a, M. Mariko a, T. Mano a, Y. Ueyama a, K. Sasaki b a Division of Oral and Maxillofacial Surgery, Department of Epithelial Intelligent and Analytical Medicine Science, Yamaguchi University Graduate School of Medicine, Japan b Division of Pathology, Department of Molecular Science and Applied Medicine, Yamaguchi University Graduate School of Medicine, Japan
We applied array-based comparative genomic hybridization for 37 primary-different oral squamous cell carcinomas (OSCCs). All materials were obtained from biopsy and excisional biopsy sample. A commercial array, MAC array Karyo 4KÒ (Macrogen, Seoul, Korea) was used in this study. This array consists of 4096 bacterial artificial chromosome (BAC) clones spotted in duplicate at an average spacing of about 1 Mb throughout the genome. In this result, gains were detected frequently at chromosomal regions of 3q29, 5p15.33, 7p22.3, 8q21.1-24.3, 9q34.3, 11q13, 16p13.3, 20p11-p12, and 20q13.3, and losses were detected frequently at chromosomal regions of 3p22, 3p14, 7q35, and 8p23.3. Gains of BAC clones locating 1.9 Mb chromosomal region of 20q13.33 and BAC clones locating 1.2 Mb chromosomal region of 16q13.3 were inversely correlated with nodal metastasis. Gains of 14 BAC clones and losses of 4 BAC clones were significantly correlated with disease-specific survival. Gains of 10 BAC clones locating 8q were inversely correlated with disease-specific survival and losses of 3 BAC clones locating 3p were associated with disease-specific survival. Two BAC clones contiguously locating 3p26.3 were correlated with disease-specific survival. Univariate Kaplan–Meier analysis showed the disease-specific survival of patients with OSCCs was negatively affected by copy number
212
Poster session III et al. / Oral Oncology Supplement 3 (2009) 201–236
doi:10.1016/j.oos.2009.06.560
P3.35. The expression of vascular endothelial growth factor A (VEGFA) and C (VEGFC) and their receptors in head and neck squamous cell carcinoma M. Macluskey a,*, S. Jones a, G. Stenhouse b, P. McLoughlin b, I. Ellis a, H. Alkhadar a a b
University of Dundee, United Kingdom Ninewells Hospital, United Kingdom
Introduction: Angiogenesis is a crucial for tumour growth, progression, and metastasis. Whilst vascular endothelial growth factor A (VEGFA) is an important angiogenic factor, vascular endothelial growth factor C (VEGFC) may stimulate the lymphatic system and facilitate lymphatic metastasis. The aim of this study was to investigate vascular endothelial growth factor A and C (VEGFA and VEGFC) and their receptors VEGFR1 and VEGFR3 in head and neck squamous cell carcinomas (HNSCC) and in nodal metastasis. Methods: Immunohistochemistry was used to identifying blood and lymphatic vessels, VEGF-A, VEGF-C and their receptors. The cohort of patients studied consisted of 22 patients with dysplastic lesions and 64 patients with cancer. Pre-treatment serum levels of VEGFA; VEGFC; VEGFR1 and VEGFR3 were determined in 63 of these HNSCC, 8 dysplasias, and in 29 normal controls using commercially available immunoassays. Results: The blood and lymphatic vessel density, VEGFA, VEGFC, VEGFR1 and VEGFR3 expression was significantly higher in HNSCC than dysplasias and significantly higher in positive nodes compared with negative nodes (p < 0.05). The mean serum levels of VEGFA and VEGFC levels were higher in the cancer group compared with both the normal and dysplastic group (p < 0.05). No association was found between any of the serum markers and nodal metastasis. High expression of VEGFA and VEGFC both within the tissue and the serum was associated with poorer survival (p < 0.05). Discussion: Disease progression from dysplasia to cancer and metastasis is accompanied by increases in both blood and lymphatic vessel density which may be facilitated not only by an up regulation of VEGFA and VEGFC but also their receptors. Systemic expression of VEGFA and VEGFC did not correlate with local expression and did not indicate metastatic disease. High expression of VEGFA and VEGFC may indicate patients with aggressive disease who may benefit from more aggressive management and may provide and chemotherapeutic target. doi:10.1016/j.oos.2009.06.561
P3.36. Overexpression of NRG1 correlating to cisplatin-resistance in oral squamous cell carcinoma S. Keiji a,*, U. Katsuhiro a, F. Kazuaki a, S. Kengo b, O. Kanae c, O. Katsunori a a
Department of Clinical Molecular Biology, Graduate School of Medicine, Chiba University, Chiba, Japan b Department of Molecular Virology, Graduate School of Medicine, Chiba University, Chiba, Japan c Division of Dentistry and Oral and Maxillofacial Surgery, Chiba University Hospital, Chiba, Japan Introduction: Resistance to cisplatin remains a major obstacle to successful treatment of oral squamous cell carcinoma (OSCC). To investigate the molecular mechanism of this resistance, we compared the gene expression profiles between the cisplatin-sensitive
SCC cell lines (Sa-3, H-1, and KB) and the cisplatin-resistant cell lines established from them (Sa-3R, H-1R, and KB-R). Methods: We analyzed the gene expression profiles of cisplatinsensitive and -resistant cell lines using the Affymetrix U133 Plus 2.0 microarray. The identified genes were used for Ingenuity Pathway Analysis Tool to identify important functional networks to cisplatin resistance. A candidate gene was selected from those genes, and then, mRNA expression of the gene was evaluated with real-time quantitative reverse transcriptase-polymerase chain reaction (qRTPCR) in both cisplatin-sensitive and -resistant cell lines. Protein expression was also examined by Western blot analysis and immunohistochemistry. In addition, to confirm the function, the gene expression was down-regulated by the small interfering RNA (siRNA) method in the cisplatin-resistant cell line. Results: Microarray analysis revealed 199 of differentially expressed genes. Network analysis was performed with these genes, and further 51 genes were shown to be crucial for the cisplatin-resistant mechanism. 51 of these genes were mapped to genetic networks and we validated the top-10 up-regulated genes by qRTPCR. NRG1 showed excellent concordance with the microarray data. Thus, we selected NRG1 for investigation. In 48 patients with OSCCs, positive immunohistochemical staining for NRG1 correlated with chemoresistance to cisplatin-based combination chemotherapy (P < 0.01). Furthermore, siRNA-directed suppression of NRG1 expression resulted in enhanced susceptibility to cisplatin. Discussion: These results suggested that overexpression of NRG1 is closely related to cisplatin-resistant mechanism in OSCC cells, and that NRG1 may be an effective chemotherapeutic target for this disease. doi:10.1016/j.oos.2009.06.562
P3.37. Loss of 3p26.3 detected by array-based comparative genomic hybridization as a predictive marker of poor prognosis in patients with oral squamous cell carcinomas K. Uchida a,*, K. Harada a, M. Mariko a, T. Mano a, Y. Ueyama a, K. Sasaki b a Division of Oral and Maxillofacial Surgery, Department of Epithelial Intelligent and Analytical Medicine Science, Yamaguchi University Graduate School of Medicine, Japan b Division of Pathology, Department of Molecular Science and Applied Medicine, Yamaguchi University Graduate School of Medicine, Japan
We applied array-based comparative genomic hybridization for 37 primary-different oral squamous cell carcinomas (OSCCs). All materials were obtained from biopsy and excisional biopsy sample. A commercial array, MAC array Karyo 4KÒ (Macrogen, Seoul, Korea) was used in this study. This array consists of 4096 bacterial artificial chromosome (BAC) clones spotted in duplicate at an average spacing of about 1 Mb throughout the genome. In this result, gains were detected frequently at chromosomal regions of 3q29, 5p15.33, 7p22.3, 8q21.1-24.3, 9q34.3, 11q13, 16p13.3, 20p11-p12, and 20q13.3, and losses were detected frequently at chromosomal regions of 3p22, 3p14, 7q35, and 8p23.3. Gains of BAC clones locating 1.9 Mb chromosomal region of 20q13.33 and BAC clones locating 1.2 Mb chromosomal region of 16q13.3 were inversely correlated with nodal metastasis. Gains of 14 BAC clones and losses of 4 BAC clones were significantly correlated with disease-specific survival. Gains of 10 BAC clones locating 8q were inversely correlated with disease-specific survival and losses of 3 BAC clones locating 3p were associated with disease-specific survival. Two BAC clones contiguously locating 3p26.3 were correlated with disease-specific survival. Univariate Kaplan–Meier analysis showed the disease-specific survival of patients with OSCCs was negatively affected by copy number