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Expression of transient receptor potential melastatin 7 in TCa8113 cells
Free Papers—Poster Presentations is a potent mitogen for epithelial cells that regulates cell proliferation, migration, survival tumour angiogenesis and invasiveness. The aim of presented study was to determine serum concentration of HGF in patients with oral squamous cell carcinoma before and after initial treatment. Methods: The investigations were carried out in a group of 40 men and 16 women. They were hospitalised at the Department of Craniomaxillofacial Surgery because of oral squamous cell carcinoma. All of them had not received any previous treatment. The control group was consisted of 20 healthy volunteers. Blood samples were collected before and four week after initial treatment. In all blood serum samples the concentrations of the HGF were determined. Results: In patients with oral squamous cell carcinoma daily oscillations of the mean values of the HGF were significantly higher than those in healthy volunteers (P < 0.001). Serum concentration of HGF was significantly higher in these patients after tumour and regional lymph nodes resection. Serum HGF level correlated positively with primary tumour stage. In our study significantly association between HGF levels and histological differentiation were not observed, but daily oscillation of HGF was higher in G3 than G2 and G1 statuses. Conclusion: Our data showed that the changes can be important pathogenic elements involved in the progression of oral squamous cell carcinoma and may be a useful marker for clinical monitoring. doi:10.1016/j.ijom.2009.03.492
P20 Serum concentration of fibroblastic growth factor in oral squamous cell carcinoma before and after surgery H. Borgiel-Marek ∗ , B. Marek, I. Niedzielska, D. Kajdaniuk Department of Craniomaxillofacial Surgery, Katowice, Silesian Medical University, Katowice, Francuska, Poland
Background and Objectives: The fibroblastic growth factor (bFGF) has been shown to induce of angiogenesis in several tumour types. The activity of bFGF during development of oral cancer has not been analysed. Numerous studies have reported correlations between prognosis and the expression status of bFGF in variety of cancer tissues. The aim of presented study was to determine serum concentration of bFGF in patients with oral squamous
cell carcinoma before and after initial treatment. Methods: Fifty six patients with oral squamous cell carcinoma who had no history of previous treatment were enrolled for the study. They were hospitalised at the Department of Craniomaxillofacial Surgery in Katowice. All patients were staged according to the TNM classification of tumours. No infection or fever was found of the patients during examinations. Blood samples were collected before and four week after initial treatment. In all blood serum samples the concentrations of the bFGF were determined. Results: In patients with oral squamous cell carcinoma daily oscillations of the mean values of the bFGF were significantly higher than those in healthy volunteers (P < 0.001). Serum concentration of bFGF was significantly higher in these patients after tumour and regional lymph nodes resection. Serum bFGF levels were not correlated with primary tumour stage. In our study significantly association between bFGF levels and histological differentiation were not observed, but daily oscillation of bFGF was higher in G3 than G2 and G1 statuses, especially after surgery. Conclusion: Our data showed that the changes can be important pathogenic elements involved in the progression of oral squamous cell carcinoma. doi:10.1016/j.ijom.2009.03.493
P21 A prospective study of jaw odontogenic keratocyst treated by multidisciplinary sequential treatment J. Sun ∗ , J. Wu, N. Zhou, N. Meng, Y. Lin College of Stomatology, Guangxi Medical University, Nanning City, Guangxi, China
Background and Objectives: To analyse and evaluate the effects of multidisciplinary sequential treatment in the dealing with jaw odontogenic keratocyst (OKC), and establish the rule of treatment with jaw OKC. Methods: The enrolment of the patients in accordance with criteria which the long diameter of OKC is more than 3.5 cm. 56 patients with OKC from June 2001 to June 2008 were included in this study. All patients were randomly assigned to multidisciplinary sequential treatment group (MST, 28 cases) or control group (CON, 28 cases). The MST group was treated with decompression, scaling of the capsule wall
537
of OKC, cutting and grinding of sclerotin of bone cavum, cauterising bone cavum by using carbolic acid. The CON group was treated with scaling of the OKC capsule wall and cauterising bone cavum by using carbolic acid. All date were recorded and studied prospectively, and the patients were followed up for 6 months to 5 years after treatment. Clinical, X-ray and pathological examinations were performed for evaluation of the clinical effect and the recurrence rate. Results: In MST group, the decompression remained 3 to 18 months, and the sizes of the cysts reduced gradually, bone regeneration was noticed on radiographs, bone cavum were 20–30% of original volume. After 2nd stage surgery, 27 cases were followed up and no recurrence was found. In the CON group, there were 3 patients wounds split. During the follow up period, recurrences of 3 cases were found. Conclusion: The MST is effective and feasible treatment method for jaw odontogenic keratocyst with many advantages, such as simple operating, resuming quondam facial shape, protecting the continuity of jaws and reduced the recurrence rate. doi:10.1016/j.ijom.2009.03.494
P22 Expression of transient receptor potential melastatin 7 in TCa8113 cells J.H. Ye 1,∗ , Y.N. Wu 1 , X. Pan 1 , J. Chen 1 , J. Gao 2 , C.P. Zhang 3 1 Department of Oral and Maxillofacial Surgery, School of Stomatology and Research Institute of Stomatology, Nanjing Medical University 2 Model Animal Research Center, Nanjing University, Nanjing, China 3 Department of Oral and Maxillofacial Surgery, School of Stomatology, Ninth People’s Hospital, Shanghai Jiao Tong University and Shanghai Research Institute of Stomatology, Shanghai, China
Background and Objectives: We investigated the electrophysiological properties and the expression of transient receptor potential melastatin 7 (TRPM7) in cultured human Tongue squamous cell carcinoma cell (TCa8113) in vitro. Methods: Cell culture, whole-cell patch clamp, reverse transcriptase-polymerase chain reaction (RT-PCR) and westernblotting. Results: There is increasing evidence that ion channels may be intimately involved in the cellular pathophysiology related to cancer. In this study, we investigated the
538 Free Papers—Poster Presentations presence and potential function of transient receptor potential melastatin 7 (TRPM7) channels in the growth and proliferation of TCa8113 cells, one common human oral squamous carcinoma cell lines, using a combination of patch-clamp recording, Western blotting, molecular biological techniques. In most of recorded TCa8113 cells, lowering extracellular Ca2+ induced a large non-desensitising current reminiscent of Ca2+-sensing cation current or TRPM7 current previously described in other cells. This Ca2+-sensing current can be inhibited by 2-aminoethoxydiphenyl borate (2-APB), or intracellular Mg2+, consistent with the TRPM7 current being activated. Consistent with it, analyses of RT-PCR, and Western blotting demonstrated the presences of TRPM7 mRNA or protein in these cells. Furthermore, the enhanced TRPM7 mRNA or protein was observed in the sample of Tongue squamous cell carcinoma. Conclusion: These results indicate that the functional expression of TRPM7 is characteristic feature of TCa8113 cells and that this pathway may be a potentially usefully target for therapeutic intervention. Acknowledgments. This study was supported by the National Natural Science Foundation of China (No. 30801301) and the Science and Technology Development Foundation of Nanjing Medical University (2006nydzd27). doi:10.1016/j.ijom.2009.03.495
P23 Overexpression of growth differentiation factor 15 is associated with cell proliferation in oral squamous cell carcinoma L. Zhang ∗ , X. Yang, H.Y. Pan, X.J. Zhou, J. Li, W.T. Chen, L.P. Zhong, Z.Y. Zhang Department of Oral and Maxillofacial Surgery, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
Background and Objectives: Our previous studies have successfully established an in vitro cellular carcinogenesis model and some differentially expressed genes in this model were identified by cDNA Microarray technology. Growth differentiation factor 15 (GDF15) was one of them. The expression of GDF15 was significantly elevated in many human tumours. However, whether the up-regulation of GDF15 can contribute to the carcinogenesis in oral squamous cell carcinoma (OSCC) still remains unclear.
Methods: The expression of GDF15 in OSCC cell lines and tissues specimens were investigated using real-time PCR, western blotting and immunohistochemistry staining. The cell growth, cell cycle and colony formation were observed after the siRNA interference against the constitutional GDF15 in Tca8113 cells. Results: Higher mRNA and protein expression of GDF15 were observed in OSCC cell lines compared with human immortalised oral epithelia cell line (HIOEC). Stronger immunoreactions and increased mRNA levels of GDF15 were observed in OSCC tissue specimens compared with adjacent non-cancerous tissues. Moreover, GDF15 expression status was correlated with the tumour differentiation stage. Inhibition of GDF15 expression by siRNA interference can significantly suppressed the cell growth and colony formation of Tca8113 cells, which may associated with cell cycle alteration. Conclusion: GDF15 is highly expressed in OSCC tissues and cell lines and associated with cancer cell differentiation. It plays a role in cell proliferation of OSCC. So GDF15 may be a potential biomarker for diagnosis and a target for anticancer therapy of OSCC. doi:10.1016/j.ijom.2009.03.496
P24 Midkine expression in salivary gland tumours T. Ota ∗ , K. Ota, H. Jono, H. Fujimori, M. Ueda, S. Shinriki, M. Kudo, T. Sueyoshi, Y. Ando, M. Shinohara Department of Oral and Maxillofacial Surgery, Kumamoto University, Kumamoto-shi, Kumamoto, Japan
Background and Objectives: Midkine (MK), a heparin-binding growth factor initially found as the product of a retinoic acid induced gene, has various biological activities. Recently, it has been reported that MK plays important roles in carcinogenesis and tumour progression, and is expressed in higher concentrations in various malignant tumours. The aim of this study was to investigate MK expression patterns in salivary gland tumours (SGTs), and to evaluate the correlation between MK expression and the degree of malignancy. Methods: We performed western blotting and immunohistochemistry to examine MK expression in adenoid cystic carcinoma (ACC), mucoepidormoid carcinoma (MC) and pleomorphic adenoma (PA) tissues. In addition, we performed immunohistochemistry for CD31 and measured
microvessel density (MVD), an indicator of angiogenesis. Results: Western blotting and immunohistochemistry showed that MK protein expression was significantly higher in malignant SGTs (ACC and MC) specimens than those in benign SGT (PA) [P < 0.001]. Furthermore, the MVD tended to be higher in cases which exhibited high expression of MK, indicating a significant correlation between the degree of MK expression and MVD (P < 0.001). Conclusions: MK may play important roles in malignant transformation and tumour angiogenesis in SGTs. doi:10.1016/j.ijom.2009.03.497
P25 Targeting interleukin-6 receptor inhibits human oral squamous cell carcinoma-related angiogenesis S. Shinriki ∗ , H. Jono, K. Ota, M. Ueda, M. Kudo, T. Ota, T. Sueyoshi, M. Ibusuki, Y. Yoshitake, Y. Ando, M. Shinohara Department of Oral and Maxillofacial Surgery, Kumamoto University, Kumamoto-shi, Kumamoto, Japan
Background and Objectives: It is unclear about the biological effect of interleukin-6 (IL-6) signalling in oral squamous cell carcinoma (OSCC) and whether IL-6 receptor (IL-6R)-mediated signalling could be a therapeutic target for OSCC. The aim of this study was to investigate the effects of inhibition of IL-6R-mediated signalling on OSCC progression. Methods: We evaluated expression levels of IL-6 and IL-6R mRNA in 58 OSCC tissues and 4 OSCC cell lines by real time quantitative RT-PCR. We investigated the effects of anti-IL-6R antibody on OSCC growth in vitro and in xenografts. Immunohistochemistry for phosphorylated signal transducer and activator of transcription 3 (pSTAT3), Ki-67, and CD31 and TUNEL assay were performed. Results: Expression levels of IL-6 mRNA in OSCC tissues were significantly higher than those in normal mucosal tissues. In addition, OSCC cell lines expressed higher levels of both IL-6 and IL-6R mRNA than HaCaT keratinocytes. Ablation of IL-6R function with anti-IL-6R antibody significantly reduced in vivo growth of SAS cells with a drastic reduction of STAT3 phosphorylation in tumour cells in mice. Inhibition of IL-6 signalling significantly decreased VEGF mRNA expression, microvessel density and vessel diameter in SAS
P20 Serum concentration of fibroblastic growth factor in oral squamous cell carcinoma before and after surgery H. Borgiel-Marek ∗ , B. Marek, I. Niedzielska, D. Kajdaniuk Department of Craniomaxillofacial Surgery, Katowice, Silesian Medical University, Katowice, Francuska, Poland
Background and Objectives: The fibroblastic growth factor (bFGF) has been shown to induce of angiogenesis in several tumour types. The activity of bFGF during development of oral cancer has not been analysed. Numerous studies have reported correlations between prognosis and the expression status of bFGF in variety of cancer tissues. The aim of presented study was to determine serum concentration of bFGF in patients with oral squamous
cell carcinoma before and after initial treatment. Methods: Fifty six patients with oral squamous cell carcinoma who had no history of previous treatment were enrolled for the study. They were hospitalised at the Department of Craniomaxillofacial Surgery in Katowice. All patients were staged according to the TNM classification of tumours. No infection or fever was found of the patients during examinations. Blood samples were collected before and four week after initial treatment. In all blood serum samples the concentrations of the bFGF were determined. Results: In patients with oral squamous cell carcinoma daily oscillations of the mean values of the bFGF were significantly higher than those in healthy volunteers (P < 0.001). Serum concentration of bFGF was significantly higher in these patients after tumour and regional lymph nodes resection. Serum bFGF levels were not correlated with primary tumour stage. In our study significantly association between bFGF levels and histological differentiation were not observed, but daily oscillation of bFGF was higher in G3 than G2 and G1 statuses, especially after surgery. Conclusion: Our data showed that the changes can be important pathogenic elements involved in the progression of oral squamous cell carcinoma. doi:10.1016/j.ijom.2009.03.493
P21 A prospective study of jaw odontogenic keratocyst treated by multidisciplinary sequential treatment J. Sun ∗ , J. Wu, N. Zhou, N. Meng, Y. Lin College of Stomatology, Guangxi Medical University, Nanning City, Guangxi, China
Background and Objectives: To analyse and evaluate the effects of multidisciplinary sequential treatment in the dealing with jaw odontogenic keratocyst (OKC), and establish the rule of treatment with jaw OKC. Methods: The enrolment of the patients in accordance with criteria which the long diameter of OKC is more than 3.5 cm. 56 patients with OKC from June 2001 to June 2008 were included in this study. All patients were randomly assigned to multidisciplinary sequential treatment group (MST, 28 cases) or control group (CON, 28 cases). The MST group was treated with decompression, scaling of the capsule wall
537
of OKC, cutting and grinding of sclerotin of bone cavum, cauterising bone cavum by using carbolic acid. The CON group was treated with scaling of the OKC capsule wall and cauterising bone cavum by using carbolic acid. All date were recorded and studied prospectively, and the patients were followed up for 6 months to 5 years after treatment. Clinical, X-ray and pathological examinations were performed for evaluation of the clinical effect and the recurrence rate. Results: In MST group, the decompression remained 3 to 18 months, and the sizes of the cysts reduced gradually, bone regeneration was noticed on radiographs, bone cavum were 20–30% of original volume. After 2nd stage surgery, 27 cases were followed up and no recurrence was found. In the CON group, there were 3 patients wounds split. During the follow up period, recurrences of 3 cases were found. Conclusion: The MST is effective and feasible treatment method for jaw odontogenic keratocyst with many advantages, such as simple operating, resuming quondam facial shape, protecting the continuity of jaws and reduced the recurrence rate. doi:10.1016/j.ijom.2009.03.494
P22 Expression of transient receptor potential melastatin 7 in TCa8113 cells J.H. Ye 1,∗ , Y.N. Wu 1 , X. Pan 1 , J. Chen 1 , J. Gao 2 , C.P. Zhang 3 1 Department of Oral and Maxillofacial Surgery, School of Stomatology and Research Institute of Stomatology, Nanjing Medical University 2 Model Animal Research Center, Nanjing University, Nanjing, China 3 Department of Oral and Maxillofacial Surgery, School of Stomatology, Ninth People’s Hospital, Shanghai Jiao Tong University and Shanghai Research Institute of Stomatology, Shanghai, China
Background and Objectives: We investigated the electrophysiological properties and the expression of transient receptor potential melastatin 7 (TRPM7) in cultured human Tongue squamous cell carcinoma cell (TCa8113) in vitro. Methods: Cell culture, whole-cell patch clamp, reverse transcriptase-polymerase chain reaction (RT-PCR) and westernblotting. Results: There is increasing evidence that ion channels may be intimately involved in the cellular pathophysiology related to cancer. In this study, we investigated the
538 Free Papers—Poster Presentations presence and potential function of transient receptor potential melastatin 7 (TRPM7) channels in the growth and proliferation of TCa8113 cells, one common human oral squamous carcinoma cell lines, using a combination of patch-clamp recording, Western blotting, molecular biological techniques. In most of recorded TCa8113 cells, lowering extracellular Ca2+ induced a large non-desensitising current reminiscent of Ca2+-sensing cation current or TRPM7 current previously described in other cells. This Ca2+-sensing current can be inhibited by 2-aminoethoxydiphenyl borate (2-APB), or intracellular Mg2+, consistent with the TRPM7 current being activated. Consistent with it, analyses of RT-PCR, and Western blotting demonstrated the presences of TRPM7 mRNA or protein in these cells. Furthermore, the enhanced TRPM7 mRNA or protein was observed in the sample of Tongue squamous cell carcinoma. Conclusion: These results indicate that the functional expression of TRPM7 is characteristic feature of TCa8113 cells and that this pathway may be a potentially usefully target for therapeutic intervention. Acknowledgments. This study was supported by the National Natural Science Foundation of China (No. 30801301) and the Science and Technology Development Foundation of Nanjing Medical University (2006nydzd27). doi:10.1016/j.ijom.2009.03.495
P23 Overexpression of growth differentiation factor 15 is associated with cell proliferation in oral squamous cell carcinoma L. Zhang ∗ , X. Yang, H.Y. Pan, X.J. Zhou, J. Li, W.T. Chen, L.P. Zhong, Z.Y. Zhang Department of Oral and Maxillofacial Surgery, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
Background and Objectives: Our previous studies have successfully established an in vitro cellular carcinogenesis model and some differentially expressed genes in this model were identified by cDNA Microarray technology. Growth differentiation factor 15 (GDF15) was one of them. The expression of GDF15 was significantly elevated in many human tumours. However, whether the up-regulation of GDF15 can contribute to the carcinogenesis in oral squamous cell carcinoma (OSCC) still remains unclear.
Methods: The expression of GDF15 in OSCC cell lines and tissues specimens were investigated using real-time PCR, western blotting and immunohistochemistry staining. The cell growth, cell cycle and colony formation were observed after the siRNA interference against the constitutional GDF15 in Tca8113 cells. Results: Higher mRNA and protein expression of GDF15 were observed in OSCC cell lines compared with human immortalised oral epithelia cell line (HIOEC). Stronger immunoreactions and increased mRNA levels of GDF15 were observed in OSCC tissue specimens compared with adjacent non-cancerous tissues. Moreover, GDF15 expression status was correlated with the tumour differentiation stage. Inhibition of GDF15 expression by siRNA interference can significantly suppressed the cell growth and colony formation of Tca8113 cells, which may associated with cell cycle alteration. Conclusion: GDF15 is highly expressed in OSCC tissues and cell lines and associated with cancer cell differentiation. It plays a role in cell proliferation of OSCC. So GDF15 may be a potential biomarker for diagnosis and a target for anticancer therapy of OSCC. doi:10.1016/j.ijom.2009.03.496
P24 Midkine expression in salivary gland tumours T. Ota ∗ , K. Ota, H. Jono, H. Fujimori, M. Ueda, S. Shinriki, M. Kudo, T. Sueyoshi, Y. Ando, M. Shinohara Department of Oral and Maxillofacial Surgery, Kumamoto University, Kumamoto-shi, Kumamoto, Japan
Background and Objectives: Midkine (MK), a heparin-binding growth factor initially found as the product of a retinoic acid induced gene, has various biological activities. Recently, it has been reported that MK plays important roles in carcinogenesis and tumour progression, and is expressed in higher concentrations in various malignant tumours. The aim of this study was to investigate MK expression patterns in salivary gland tumours (SGTs), and to evaluate the correlation between MK expression and the degree of malignancy. Methods: We performed western blotting and immunohistochemistry to examine MK expression in adenoid cystic carcinoma (ACC), mucoepidormoid carcinoma (MC) and pleomorphic adenoma (PA) tissues. In addition, we performed immunohistochemistry for CD31 and measured
microvessel density (MVD), an indicator of angiogenesis. Results: Western blotting and immunohistochemistry showed that MK protein expression was significantly higher in malignant SGTs (ACC and MC) specimens than those in benign SGT (PA) [P < 0.001]. Furthermore, the MVD tended to be higher in cases which exhibited high expression of MK, indicating a significant correlation between the degree of MK expression and MVD (P < 0.001). Conclusions: MK may play important roles in malignant transformation and tumour angiogenesis in SGTs. doi:10.1016/j.ijom.2009.03.497
P25 Targeting interleukin-6 receptor inhibits human oral squamous cell carcinoma-related angiogenesis S. Shinriki ∗ , H. Jono, K. Ota, M. Ueda, M. Kudo, T. Ota, T. Sueyoshi, M. Ibusuki, Y. Yoshitake, Y. Ando, M. Shinohara Department of Oral and Maxillofacial Surgery, Kumamoto University, Kumamoto-shi, Kumamoto, Japan
Background and Objectives: It is unclear about the biological effect of interleukin-6 (IL-6) signalling in oral squamous cell carcinoma (OSCC) and whether IL-6 receptor (IL-6R)-mediated signalling could be a therapeutic target for OSCC. The aim of this study was to investigate the effects of inhibition of IL-6R-mediated signalling on OSCC progression. Methods: We evaluated expression levels of IL-6 and IL-6R mRNA in 58 OSCC tissues and 4 OSCC cell lines by real time quantitative RT-PCR. We investigated the effects of anti-IL-6R antibody on OSCC growth in vitro and in xenografts. Immunohistochemistry for phosphorylated signal transducer and activator of transcription 3 (pSTAT3), Ki-67, and CD31 and TUNEL assay were performed. Results: Expression levels of IL-6 mRNA in OSCC tissues were significantly higher than those in normal mucosal tissues. In addition, OSCC cell lines expressed higher levels of both IL-6 and IL-6R mRNA than HaCaT keratinocytes. Ablation of IL-6R function with anti-IL-6R antibody significantly reduced in vivo growth of SAS cells with a drastic reduction of STAT3 phosphorylation in tumour cells in mice. Inhibition of IL-6 signalling significantly decreased VEGF mRNA expression, microvessel density and vessel diameter in SAS