Pseudomonas aeruginosa로부터 FPLC를 이용한 protease IV의 분리와 그것의 사람구강편평상피암종 세포에 미치는 세포독성효과
Extraction of protease IV from Pseudomonas aeruginosa by FPLC and its Cytotoxic effect on oral squamous carcinoma cells
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Oral squamous carcinoma (OSC) cell is responsib1e for the majority of malignancies in the oral cavity. The majority of oral OSC could be treated by a wide surgic외 resection and/or radiation therapy but showed the lowest 5- year survival rate among the major '- cancers. After reports on regression of cancer in humans and animals infected with microbial pathogens date back more than 100 years. much effort has been spent over the years in developing wild-type or attenuated bacterial and purified bacterial proteins for the treatment of cancer. Pseudomonas aeruginosa has extracell띠ar virulence factors such as exotoxin A. exoenzyme S. elastase. protease IV (PIV) and alkaline protease. Pseudomonas aerugino.sa has 아en repoπ.ed to induce apoptosis in epithelial cells. endothelial ceJ1s and macrophage. My laboratory found that the extraceJ1uar products of Pseudo.monas aerugino.있 (EPPA) have a preferential cytotoxic effect on human or외 squamous carcinoma cells compared with human osteosarcoma cells such as HOS and MG63. In order to find the key agent which has such a effect. 1 and collaborators have extracted many kinds of proteins by various methods and studied its cytotoxic effect on OSC cells, 1 obseπed many peaks of proteins from EPPA through FPLC 때d performed to extract 26 kDa PIV from them Because the pattern of PIV peak was distinct from other peaks. it was convenient to extract PIV through FPLC without using the gene c10lÙng‘ When 1 perform protein e1ectrophoresis with extracted PIV. no other bands found except 26 kDa PIV band. suggesting that extracted PIV was high1y pure‘ AJthough the cytotoxic effect of extracted PIV was lower than exotoxin-A and Las B elastase from Pseudo.mo.nas aerulIino.sa. PIV 1ed OSC cells to apoptosis bv means of Hoechst stain
Abstract
Oral squamous carcinoma (OSC) cell is responsib1e for the majority of malignancies in the oral cavity. The majority of oral OSC could be treated by a wide surgic외 resection and/or radiation therapy but showed the lowest 5- year survival rate among the major '- cancers. After reports on regression of cancer in humans and animals infected with microbial pathogens date back more than 100 years. much effort has been spent over the years in developing wild-type or attenuated bacterial and purified bacterial proteins for the treatment of cancer. Pseudomonas aeruginosa has extracell띠ar virulence factors such as exotoxin A. exoenzyme S. elastase. protease IV (PIV) and alkaline protease. Pseudomonas aerugino.sa has 아en repoπ.ed to induce apoptosis in epithelial cells. endothelial ceJ1s and macrophage. My laboratory found that the extraceJ1uar products of Pseudo.monas aerugino.있 (EPPA) have a preferential cytotoxic effect on human or외 squamous carcinoma cells compared with human osteosarcoma cells such as HOS and MG63. In order to find the key agent which has such a effect. 1 and collaborators have extracted many kinds of proteins by various methods and studied its cytotoxic effect on OSC cells, 1 obseπed many peaks of proteins from EPPA through FPLC 때d performed to extract 26 kDa PIV from them Because the pattern of PIV peak was distinct from other peaks. it was convenient to extract PIV through FPLC without using the gene c10lÙng‘ When 1 perform protein e1ectrophoresis with extracted PIV. no other bands found except 26 kDa PIV band. suggesting that extracted PIV was high1y pure‘ AJthough the cytotoxic effect of extracted PIV was lower than exotoxin-A and Las B elastase from Pseudo.mo.nas aerulIino.sa. PIV 1ed OSC cells to apoptosis bv means of Hoechst stain