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Expression of the urokinase receptor regulates focal adhesion assembly and cell migration in adenoid cystic carcinoma cells
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Int. J. Oral Maxillofac. Surg. 2005; 34 (Supplement 1): $ 1 - $ 1 8 1
[-P'2-'~ EXPRESSION OF THE UROKINASE RECEPTOR REGULATES FOCAL ADHESION ASSEMBLY AND CELL MIGRATION IN ADENOID CYSTIC CARCINOMA CELLS
[-~-'~
T. Sugiura, S. Abu-Ali, M. Takahashi, K. Shirasuna. Department of Oral
Surgery, School of Dentistry, Mayor University, Santiago de Chile, Chile
and Maxillofacial Surgery, Graduate School of Dental Science, Kyushu University, Fukuoka, Japan Adenoid cystic carcinoma (AdCC) grows slowly, but spreads relentlessly into adjacent tissues; it shows a special proclivity for invading nerves and endothelial sheath. The frequencies of recurrence and distant metastasis of AdCC are very high. To elucidate the invasion mechanisms of AdCC cells, this study focused on collagen-induced cell migration and found an important role of uPAR in focal adhesion assembly and migration. AdCC cell lines (ACCS and ACCT), oral squamous cell carcinoma (SCC) line NA AdCC cell lines (ACCS and ACCT) showed higher migration responses and adhesion to the extracellular matrix (ECM), especially types I and IV collagen, than did SCC cell line NA. The response to collagens was largely and exclusively inhibited by anti-~2 integrin antibody. Moreover, AdCC cell lines expressed higher surface levels of urokinase-type plasminogen activator receptor (uPAR) than did SCC cell lines. When AdCC cells were plated on collagen, the surface level of uPAR was increased, and numerous focal adhesions consisting of uPAR, vinculin, and paxillin were assembled; whereas collagen-stimulated SCC cell counterparts or AdCC cells plated on other types of ECM, such as fibronectin, failed to assemble such definite focal adhesions. In order to elucidate the association of uPAR with collagen-induced events, an ACCS-AS cell line transfected with a vector expressing antisense uPAR RNA was established and shown to have reduced uPAR (about 10% that of parental ACCS at both the protein and mRNA levels). ACCSAS showed a strong reduction of collagen-stimulated migration and focal adhesion assembly of ~2 integrin, vinculin, and paxillin. These findings suggest that AdCC has a proclivity for migrating to type I and IV collagens due to the overexpression of uPAR, which plays a key role in focal adhesion assembly and migration.
[-P'2"-~ MRNA EXPRESSION OF CK 5, CEA & SURVIVIN IN LYMPH NODE AS PROGNOSIS INDICATORS IN ORAL SQUAMOUS CELL CARCINOMA K.-Y. Lim, M.-J. Kim, M. Hoon, J.-Y. Paeng, J.-H. Park. College of
Dentistry, Seoul National University, Seoul, South Korea The purpose of this study was to investigate mRNA expression of cytokeratin (CK) 5, carcinoembryonic antigen (CEA) and survivin in lymph nodes in patients with OSCC and compare their expression with clinico-pathological prognostic factor such as, gender, age, T score, clinical stage, differentiation, proliferation, invasion index, and lymph node metastasis We ecamined 10 normal gingival tissues, 11 OSCC tumor tissue and 20 lymph nodes of OSCC patients to evaluate expression of CK 5, CEA and survivin by RT-PCR and immunohistochemistry. Clinical information including age, T score, gender, and TNM stage from 97 patients with OSCC were reivewed. Tumor differentiation & invasion index (by H&E), proliferating cell nuclear antigen (PCNA) (by immunohistochemistry) were examined. 281 lymph node were examined by RT-PCR analysis. Survival curves were estimated by the Kaplan-Meier method and normal or numeric variable influence on survival was studied by univariate and mutivariate Regression analysis (Cox proportional hazards model). Results: In normal gingival tissue, CK 5(100%), CEA (10%) and survivin (0%) were expressed. In OSCC tissues, CK 5 (100%), CEA (9-18%) and survivin (82-91%) were expressed. In metastatic lymph nodes, CK 5(100%), CEA (16.7%) and survivin (100%) were expressed, but on the other hand, in histologically negative lymph nodes, CK 5, CEA and survivin were partially expressed immunohistochemistry (0£21.4%), by RT-PCR (7.1-42.9%). Therefore RT-PCR showed higher sensitivity than immunohistochemistry. We could get following results by statistical analysis rate fo 97 patients. 1. Lynph node survivin mRNA expressed and clinical stage were independent OSCC prognosis factors. 2. Lymph nod CK 5 and survivin mRNA expression had significant effects on survival rate. 3. Stage, T score and lymph node metastasis significantly influenced survival rate. 4. CEA mRNA expression, gender, age, differentiation, invasion index and PCNA scores had no significant effect on survival rate. we can predict micrometastasis of OSCC in case of mRNA expression of CK 5 and survivin in lymph node. Lymph nod survivin expression might provide predictive information for OSCC patient survival.
PERIPHERAL ODONTOGENIC FIBROMA. CASE REPORT AND REVIEW OF THE LITERATURE
C. Montini, L. Castellon, A. Basili, R. Farina, G. Laissle. Dept. of Oral The peripheral odontogenic fibroma (WHO type) is a relatively rare, benign, unencapsulated, gingival mass of fibrous connective tissue. It is considered to be the extraosseous counterpart of the central odontogenic fibroma. Histologically it is characterized by a fibrous or fibromyxomatous proliferation that contains varying amounts of odontogenic or presumed odontogenic epithelium. Usually it haas not radiographic features. The purpose of this article is to reposrt a case of a peripheral odontogenic fibroma in the mandible, in a 24-year-old male, an uncommon location. We also make a review of the literature to compare the clinical and histological aspects of this type of lesion. [-~-'~
A CASE OF OSTEOSARCOMA OF MANDIBULAR CONDYLAR PROCESS
K. Yago 1 , S. Asanami 1 , R. Ryu 1 , K. Nishiyama 1 , T. Nakagawa1, Y. Tanaka2 , K. Yoshida3. 1Department of Dentistry and Oral Surgery,
School of Medicine, Keio University, Japan; 2Clinical Laboratory, Surgical Pathology, Ichikawa General Hospital Tokyo Dental College, Japan; 3Department of Brain Surgery, School of Medicine, Keio University, Japan Osteosarcoma of the jaw is uncommon. We report a rare case of osteosarcoma of mandibular condylar process. We treated a patient with osteosarcoma in mandibular condylar process by combination therapy with pre-and post surgical chemotherapy using high-dose methotrexate citrovorum factor (HD-MTX CF) resume. A 27-year-old female visited our hospital with a complaint of trismus and paresthesia of the lower lip. Panoramaic radiograph revealed irregular bone formation and a sunray appearance in the right mandibular condylar process. CT scan demonstrated measuring 4 x 3 . 5 x 3 cm mass that might be invading into the brain. Bone scintigraphic scans showed an area of increased intake of radiolabeled technetium. Histological diagnosis of a biopsy specimen indicated the lesion to be an osteosarcoma. It showed osteoblastic differentiation. The distribution of grade according to the histologic evaluation of malignancy by Broders was grade 1. We perfomed chemotherapy with a protocol NECO-95, HD-MTX CF therapy in combination with adriamycine (ADM) and cisplatin (CDDP). The patient received HD-MTX CF therapy three times in accordance NECO-95 protocol before radical operation. MTX is administered at a dose of 12g/m2. The peak MTX levels of the serum stayed greater than 1000pmol/L that was regarded as the effective range. The side effect of MTX were limited to a nausea, a vomiting, and slight leucopenia, and critical adverse effects by MTX were not observed. The patient underwent wide excision of the mandible with resection of the middle part of the skull base. The patient received HD-MTX CF therapy eight times as an ajuvant chemotherapy after the operation. There was no evidence of tumor recurrence and metastasis while three years after the radical operation. This case indicated that the chemotherapy with systemic HD-MTX is effective for osteosarcoma of the jaw and we could completely resect the lesion in cooperarion with brain surgeons. [-~-'~
THE EFFECT OF BETA EMITTING RADIONUCLIDE (HOLMIUM)-CHITOSAN COMPLEX ON ORAL CANCER: IN VIVO AND IN VITRO STUDY
J.-'~ Paeng, M.-J. Kim, B.-C. Shin, H. Myoung. Dept. of Oral and
Maxillofacial Surgery, College of Dentistry, Seoul National University, Seoul, and Korea Atomic Energy Research Institute, Deajeon, South Korea The purpose of this study was to evaluate the effect of beta emitting radionuclide Ho-166 and its complex with chitosan on oral cancer Radiosensitivities of oral cancer cell lines(KB, HSC-3) were evaluated with MTT assay. Apoptosis of cells after irradiation from Ho-166 was measured with TUNEL assay and flow cytometry. We induced oral cancer mass in the flank of 20 Balb/c/nu mice with inoculating oral cancer cell line (KB cell line) subcutaneously. We prepared the chitosan solutions with high molecular weight chitosan (Sigma Aldrich, USA). After mixing with 166Ho(NO3)35H20, the Ho-chitosan complex was injected intratumorally at the dose of 185 Mbq in 0.2ml. To the control group, saline
Int. J. Oral Maxillofac. Surg. 2005; 34 (Supplement 1): $ 1 - $ 1 8 1
[-P'2-'~ EXPRESSION OF THE UROKINASE RECEPTOR REGULATES FOCAL ADHESION ASSEMBLY AND CELL MIGRATION IN ADENOID CYSTIC CARCINOMA CELLS
[-~-'~
T. Sugiura, S. Abu-Ali, M. Takahashi, K. Shirasuna. Department of Oral
Surgery, School of Dentistry, Mayor University, Santiago de Chile, Chile
and Maxillofacial Surgery, Graduate School of Dental Science, Kyushu University, Fukuoka, Japan Adenoid cystic carcinoma (AdCC) grows slowly, but spreads relentlessly into adjacent tissues; it shows a special proclivity for invading nerves and endothelial sheath. The frequencies of recurrence and distant metastasis of AdCC are very high. To elucidate the invasion mechanisms of AdCC cells, this study focused on collagen-induced cell migration and found an important role of uPAR in focal adhesion assembly and migration. AdCC cell lines (ACCS and ACCT), oral squamous cell carcinoma (SCC) line NA AdCC cell lines (ACCS and ACCT) showed higher migration responses and adhesion to the extracellular matrix (ECM), especially types I and IV collagen, than did SCC cell line NA. The response to collagens was largely and exclusively inhibited by anti-~2 integrin antibody. Moreover, AdCC cell lines expressed higher surface levels of urokinase-type plasminogen activator receptor (uPAR) than did SCC cell lines. When AdCC cells were plated on collagen, the surface level of uPAR was increased, and numerous focal adhesions consisting of uPAR, vinculin, and paxillin were assembled; whereas collagen-stimulated SCC cell counterparts or AdCC cells plated on other types of ECM, such as fibronectin, failed to assemble such definite focal adhesions. In order to elucidate the association of uPAR with collagen-induced events, an ACCS-AS cell line transfected with a vector expressing antisense uPAR RNA was established and shown to have reduced uPAR (about 10% that of parental ACCS at both the protein and mRNA levels). ACCSAS showed a strong reduction of collagen-stimulated migration and focal adhesion assembly of ~2 integrin, vinculin, and paxillin. These findings suggest that AdCC has a proclivity for migrating to type I and IV collagens due to the overexpression of uPAR, which plays a key role in focal adhesion assembly and migration.
[-P'2"-~ MRNA EXPRESSION OF CK 5, CEA & SURVIVIN IN LYMPH NODE AS PROGNOSIS INDICATORS IN ORAL SQUAMOUS CELL CARCINOMA K.-Y. Lim, M.-J. Kim, M. Hoon, J.-Y. Paeng, J.-H. Park. College of
Dentistry, Seoul National University, Seoul, South Korea The purpose of this study was to investigate mRNA expression of cytokeratin (CK) 5, carcinoembryonic antigen (CEA) and survivin in lymph nodes in patients with OSCC and compare their expression with clinico-pathological prognostic factor such as, gender, age, T score, clinical stage, differentiation, proliferation, invasion index, and lymph node metastasis We ecamined 10 normal gingival tissues, 11 OSCC tumor tissue and 20 lymph nodes of OSCC patients to evaluate expression of CK 5, CEA and survivin by RT-PCR and immunohistochemistry. Clinical information including age, T score, gender, and TNM stage from 97 patients with OSCC were reivewed. Tumor differentiation & invasion index (by H&E), proliferating cell nuclear antigen (PCNA) (by immunohistochemistry) were examined. 281 lymph node were examined by RT-PCR analysis. Survival curves were estimated by the Kaplan-Meier method and normal or numeric variable influence on survival was studied by univariate and mutivariate Regression analysis (Cox proportional hazards model). Results: In normal gingival tissue, CK 5(100%), CEA (10%) and survivin (0%) were expressed. In OSCC tissues, CK 5 (100%), CEA (9-18%) and survivin (82-91%) were expressed. In metastatic lymph nodes, CK 5(100%), CEA (16.7%) and survivin (100%) were expressed, but on the other hand, in histologically negative lymph nodes, CK 5, CEA and survivin were partially expressed immunohistochemistry (0£21.4%), by RT-PCR (7.1-42.9%). Therefore RT-PCR showed higher sensitivity than immunohistochemistry. We could get following results by statistical analysis rate fo 97 patients. 1. Lynph node survivin mRNA expressed and clinical stage were independent OSCC prognosis factors. 2. Lymph nod CK 5 and survivin mRNA expression had significant effects on survival rate. 3. Stage, T score and lymph node metastasis significantly influenced survival rate. 4. CEA mRNA expression, gender, age, differentiation, invasion index and PCNA scores had no significant effect on survival rate. we can predict micrometastasis of OSCC in case of mRNA expression of CK 5 and survivin in lymph node. Lymph nod survivin expression might provide predictive information for OSCC patient survival.
PERIPHERAL ODONTOGENIC FIBROMA. CASE REPORT AND REVIEW OF THE LITERATURE
C. Montini, L. Castellon, A. Basili, R. Farina, G. Laissle. Dept. of Oral The peripheral odontogenic fibroma (WHO type) is a relatively rare, benign, unencapsulated, gingival mass of fibrous connective tissue. It is considered to be the extraosseous counterpart of the central odontogenic fibroma. Histologically it is characterized by a fibrous or fibromyxomatous proliferation that contains varying amounts of odontogenic or presumed odontogenic epithelium. Usually it haas not radiographic features. The purpose of this article is to reposrt a case of a peripheral odontogenic fibroma in the mandible, in a 24-year-old male, an uncommon location. We also make a review of the literature to compare the clinical and histological aspects of this type of lesion. [-~-'~
A CASE OF OSTEOSARCOMA OF MANDIBULAR CONDYLAR PROCESS
K. Yago 1 , S. Asanami 1 , R. Ryu 1 , K. Nishiyama 1 , T. Nakagawa1, Y. Tanaka2 , K. Yoshida3. 1Department of Dentistry and Oral Surgery,
School of Medicine, Keio University, Japan; 2Clinical Laboratory, Surgical Pathology, Ichikawa General Hospital Tokyo Dental College, Japan; 3Department of Brain Surgery, School of Medicine, Keio University, Japan Osteosarcoma of the jaw is uncommon. We report a rare case of osteosarcoma of mandibular condylar process. We treated a patient with osteosarcoma in mandibular condylar process by combination therapy with pre-and post surgical chemotherapy using high-dose methotrexate citrovorum factor (HD-MTX CF) resume. A 27-year-old female visited our hospital with a complaint of trismus and paresthesia of the lower lip. Panoramaic radiograph revealed irregular bone formation and a sunray appearance in the right mandibular condylar process. CT scan demonstrated measuring 4 x 3 . 5 x 3 cm mass that might be invading into the brain. Bone scintigraphic scans showed an area of increased intake of radiolabeled technetium. Histological diagnosis of a biopsy specimen indicated the lesion to be an osteosarcoma. It showed osteoblastic differentiation. The distribution of grade according to the histologic evaluation of malignancy by Broders was grade 1. We perfomed chemotherapy with a protocol NECO-95, HD-MTX CF therapy in combination with adriamycine (ADM) and cisplatin (CDDP). The patient received HD-MTX CF therapy three times in accordance NECO-95 protocol before radical operation. MTX is administered at a dose of 12g/m2. The peak MTX levels of the serum stayed greater than 1000pmol/L that was regarded as the effective range. The side effect of MTX were limited to a nausea, a vomiting, and slight leucopenia, and critical adverse effects by MTX were not observed. The patient underwent wide excision of the mandible with resection of the middle part of the skull base. The patient received HD-MTX CF therapy eight times as an ajuvant chemotherapy after the operation. There was no evidence of tumor recurrence and metastasis while three years after the radical operation. This case indicated that the chemotherapy with systemic HD-MTX is effective for osteosarcoma of the jaw and we could completely resect the lesion in cooperarion with brain surgeons. [-~-'~
THE EFFECT OF BETA EMITTING RADIONUCLIDE (HOLMIUM)-CHITOSAN COMPLEX ON ORAL CANCER: IN VIVO AND IN VITRO STUDY
J.-'~ Paeng, M.-J. Kim, B.-C. Shin, H. Myoung. Dept. of Oral and
Maxillofacial Surgery, College of Dentistry, Seoul National University, Seoul, and Korea Atomic Energy Research Institute, Deajeon, South Korea The purpose of this study was to evaluate the effect of beta emitting radionuclide Ho-166 and its complex with chitosan on oral cancer Radiosensitivities of oral cancer cell lines(KB, HSC-3) were evaluated with MTT assay. Apoptosis of cells after irradiation from Ho-166 was measured with TUNEL assay and flow cytometry. We induced oral cancer mass in the flank of 20 Balb/c/nu mice with inoculating oral cancer cell line (KB cell line) subcutaneously. We prepared the chitosan solutions with high molecular weight chitosan (Sigma Aldrich, USA). After mixing with 166Ho(NO3)35H20, the Ho-chitosan complex was injected intratumorally at the dose of 185 Mbq in 0.2ml. To the control group, saline