- Email: [email protected]
Poster 023: The Regulative Role of Human Papillomavirus in Malignant Transformation of Oral Epithelial Cells
Scientific Poster Session
POSTER 023
POSTER 024
The Regulative Role of Human Papillomavirus in Malignant Transformation of Oral Epithelial Cells
Decreased Expression of Annexin A1 Related With Pathologic Differentiation Grade in Oral Squamous Cell Carcinoma
Rong-tao Yuan, PhD, Affiliated Hospital, Medical College, Qingdao University, Department of Oral and Maxillofacial Surgery, 16 Jiangsu Road, Qingdao, Shandong, 266003, China (Zhou XJ; Pan HY; Zhang P; Chen WT; Zhang ZY)
Lai-ping Zhong, DDS, MD, PhD, Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, No.639 Zhizaoju Rd, Shanghai, 200011, China (Zhang L; Yang X; Wei KJ; Zhou XJ; Pan HY; Zhang ZY)
Abstract Objective: To investigate the regulative role of human papillomavirus (HPV) in the process of transformation and malignancy of oral epithelial cells and to search its functionary mechanism in the carcinogenesis of epithelial cells through detecting the relation between HPV16 E6 and cell cycle proteins and telomerase in the different period of oral mucosa epithelial cells transformation. Methods: Oral mucosa epithelial cells were infected with LXSN HPV16 E6 retrovirus in vitro. PCR, Southern blot, and immunocytochemical staining were performed to detect the expression of the HPV16 E6 in the host cells. Detections of Rb protein were carried out with Western blot method in the normal epithelial cells, HPV16 E6 transformed epithelial cells, HPV16 E6/E7 immortalized epithelial cells, and Tca8113 tongue cancer cells. Activation of telomerase was inspected with PCR-TRAP ELISA. Results: It was identified that HPV16 E6 was integrated into the genome of host cells and expressed in the cytoplasm. The expression of Rb protein in normal epithelial cells was twofold to triplication of those in HPV16 E6 transformed cells. Those in HPV16 E6/E7 immortalized cells and tongue cancer cells were sharply decreased at the mean time. Telomerase in HPV16 E6 transformed cells was activated while the activity of telomerase in normal epithelial cells was not detected. The activity of telomerase in HPV16 E6/E7 immortalized cells was significantly higher than those in HPV16 E6 transformed cells (p⬍0.05). Tongue cancer cells had the highest activity of telomerase (p⬍0.05). Conclusion: While HPV16 was transfected into normal oral mucosa epithelial cells, it integrated into the genome of host cells and expressed in protein level. HPV16 decreased the expression of Rb protein and partially inactivated the negative regulation of growth and proliferation of epithelial cells, which extended the life span of the cells. Telomerase was activated in HPV16 transformed cells. While it got high levels, epithelial cells were immortalized or malignant transformation. HPV16 played multiple roles in the process of transformation and malignancy of oral epithelial cells.
Background: Our previous studies have successfully established an in vitro cellular carcinogenesis model of oral squamous cell carcinoma (OSCC), including a human immortalized oral epithelia cell (HIOEC) line and a cancerous cell line (HB96) from HIOEC by induction with benzo(a)pyrene. Further differentially expressed proteins in this cellular carcinogenesis model were identified using comparative proteomic analysis, which included Annexin A1. Objective: The aim of this study was to describe the significant decreased expression of Annexin A1 in the HB96 cells compared with the HIOECs, and to investigate the potential usefulness of Annexin A1 in OSCC. Methods: The HIOECs and HB96 cells in the in vitro cellular carcinogenesis model of OSCC, and other OSCC cell lines of Tca8113, TSCC, CAL27, OSC, and NT were used. Thirty pairs of surgical tissue samples from 30 primary OSCC patients, including cancerous tissue and adjacent non-malignant epithelium, were collected during surgery. The Annexin A1 protein expression and mRNA level in the HIOECs and OSCC cell lines were detected using Western blotting and real-time PCR, respectively; and those in the cancerous and adjacent non-malignant epithelia from OSCC patients were detected using immunohistochemistry and real-time PCR, respectively. All data were analyzed using the statistical software of SPSS10.0 for Windows. Results: Decreased Annexin A1 mRNA level and protein expression were also found in all OSCC cell lines except the Annexin A1 mRNA level in two cell lines. Decreased Annexin A1 mRNA level and protein expression were found in the cancerous tissues compared with the adjacent non-malignant epithelia. Annexin A1 mRNA level and protein expression in cancerous tissues were both found correlating with the pathologic differentiation grade of cancerous tissues, a lower Annexin A1 mRNA level or protein expression in the cancerous tissue indicating a poorer pathologic differentiation grade. Conclusion: From these results, we suggest that decreased expression of Annexin A1 contributes to the cancerous progression of OSCC, and Annexin A1 is a potential biomarker for pathologic differentiation grade of OSCC.
82
AAOMS • 2008
POSTER 023
POSTER 024
The Regulative Role of Human Papillomavirus in Malignant Transformation of Oral Epithelial Cells
Decreased Expression of Annexin A1 Related With Pathologic Differentiation Grade in Oral Squamous Cell Carcinoma
Rong-tao Yuan, PhD, Affiliated Hospital, Medical College, Qingdao University, Department of Oral and Maxillofacial Surgery, 16 Jiangsu Road, Qingdao, Shandong, 266003, China (Zhou XJ; Pan HY; Zhang P; Chen WT; Zhang ZY)
Lai-ping Zhong, DDS, MD, PhD, Ninth People’s Hospital, School of Medicine, Shanghai Jiao Tong University, No.639 Zhizaoju Rd, Shanghai, 200011, China (Zhang L; Yang X; Wei KJ; Zhou XJ; Pan HY; Zhang ZY)
Abstract Objective: To investigate the regulative role of human papillomavirus (HPV) in the process of transformation and malignancy of oral epithelial cells and to search its functionary mechanism in the carcinogenesis of epithelial cells through detecting the relation between HPV16 E6 and cell cycle proteins and telomerase in the different period of oral mucosa epithelial cells transformation. Methods: Oral mucosa epithelial cells were infected with LXSN HPV16 E6 retrovirus in vitro. PCR, Southern blot, and immunocytochemical staining were performed to detect the expression of the HPV16 E6 in the host cells. Detections of Rb protein were carried out with Western blot method in the normal epithelial cells, HPV16 E6 transformed epithelial cells, HPV16 E6/E7 immortalized epithelial cells, and Tca8113 tongue cancer cells. Activation of telomerase was inspected with PCR-TRAP ELISA. Results: It was identified that HPV16 E6 was integrated into the genome of host cells and expressed in the cytoplasm. The expression of Rb protein in normal epithelial cells was twofold to triplication of those in HPV16 E6 transformed cells. Those in HPV16 E6/E7 immortalized cells and tongue cancer cells were sharply decreased at the mean time. Telomerase in HPV16 E6 transformed cells was activated while the activity of telomerase in normal epithelial cells was not detected. The activity of telomerase in HPV16 E6/E7 immortalized cells was significantly higher than those in HPV16 E6 transformed cells (p⬍0.05). Tongue cancer cells had the highest activity of telomerase (p⬍0.05). Conclusion: While HPV16 was transfected into normal oral mucosa epithelial cells, it integrated into the genome of host cells and expressed in protein level. HPV16 decreased the expression of Rb protein and partially inactivated the negative regulation of growth and proliferation of epithelial cells, which extended the life span of the cells. Telomerase was activated in HPV16 transformed cells. While it got high levels, epithelial cells were immortalized or malignant transformation. HPV16 played multiple roles in the process of transformation and malignancy of oral epithelial cells.
Background: Our previous studies have successfully established an in vitro cellular carcinogenesis model of oral squamous cell carcinoma (OSCC), including a human immortalized oral epithelia cell (HIOEC) line and a cancerous cell line (HB96) from HIOEC by induction with benzo(a)pyrene. Further differentially expressed proteins in this cellular carcinogenesis model were identified using comparative proteomic analysis, which included Annexin A1. Objective: The aim of this study was to describe the significant decreased expression of Annexin A1 in the HB96 cells compared with the HIOECs, and to investigate the potential usefulness of Annexin A1 in OSCC. Methods: The HIOECs and HB96 cells in the in vitro cellular carcinogenesis model of OSCC, and other OSCC cell lines of Tca8113, TSCC, CAL27, OSC, and NT were used. Thirty pairs of surgical tissue samples from 30 primary OSCC patients, including cancerous tissue and adjacent non-malignant epithelium, were collected during surgery. The Annexin A1 protein expression and mRNA level in the HIOECs and OSCC cell lines were detected using Western blotting and real-time PCR, respectively; and those in the cancerous and adjacent non-malignant epithelia from OSCC patients were detected using immunohistochemistry and real-time PCR, respectively. All data were analyzed using the statistical software of SPSS10.0 for Windows. Results: Decreased Annexin A1 mRNA level and protein expression were also found in all OSCC cell lines except the Annexin A1 mRNA level in two cell lines. Decreased Annexin A1 mRNA level and protein expression were found in the cancerous tissues compared with the adjacent non-malignant epithelia. Annexin A1 mRNA level and protein expression in cancerous tissues were both found correlating with the pathologic differentiation grade of cancerous tissues, a lower Annexin A1 mRNA level or protein expression in the cancerous tissue indicating a poorer pathologic differentiation grade. Conclusion: From these results, we suggest that decreased expression of Annexin A1 contributes to the cancerous progression of OSCC, and Annexin A1 is a potential biomarker for pathologic differentiation grade of OSCC.
82
AAOMS • 2008