Molecular analyses of BP180 in regulation of prostate cancer cell spreading and migration activities

• Main Authors: Chia-Cheng Chang, 張家誠
• Other Authors: 宋賢穎
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/51242154452352026130

碩士 === 中國醫藥大學 === 癌症生物學研究所碩士班 === 101 === Backgrounds: 　　Collagen XVII (BP180), a major component of the hemidesmosome is critical in the maintenance of cell adhesion. Clinical evidences indicated mutation or loss of BP180 leaded to subepidermal tissue separation. In addition, BP180 has been shown to play an important role of cancer migration and metastasis. However, the detail mechanism is still not clear. Hence, we aimed to investigate the mechanisms of BP1 80 in regulation of cancer migration and invasion. Materials and Methods: 　　We first knockdown the expression levels of BP180 in PC3 cell lines. Lentiviral knockdown of BP180 was constructed. Real-time PCR was used to confirm knockdown efficiency. Cell adhesion, spreading and migration assay was performed. ADAM9 followed by analysis of BP180, expression was performed. Protein Integrin β4 degradation of hemidesmosome components after knockdown of ADAM9 was determined by cycloheximide treated prostate cancer cells. Results: 　　We noticed the differential expression pattern of BP180 across different cell lines, with strong expression in prostate cancer cells and lowest in lung cancer carcinoma, especially in PC3 cell. Inhibition of cell migration activities can be detected in prostate cancer cell line, PC3 after knockdown of BP180 expression. In addition, we also observed decreased cell spreading, as well as haptotactic migration activities on matrices. Furthermore, we confirmed the interaction of ADAM9 and Integrin β4 and observed the endocytosis and degradation. Furthermore, shedding of BP180 by ADAM9 reversed had been observed. But MMP inhibitor treated resulted in BP180 expression decreased. Conclusion: 　　Our results indicated ADAM9 regulated hemidesmosome endocytosis and degradation by direct interaction with BP180. And knockdown of BP180 inhibited cell migration and spreading activities by upholding constitutive expression levels of hemidesmosome complex in prostate cancer cells. Therefore, it is plausible to hypothesize the therapeutic strategy by blocking ADAM9-BP180 interaction could inhibit cancer cell metastasis activities.