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Objective: The purpose of this study was to determine the mechanism of action of clusterin-known as a chemo-resistance protein-by analyzing its binding with chemotherapeutic agents and elucidating its relation with drug resistance. Methods: Chemotherapeutic agents were diluted with coating buffer an...

Objective: The purpose of this study was to determine the mechanism of action of clusterin-known as a chemo-resistance protein-by analyzing its binding with chemotherapeutic agents and elucidating its relation with drug resistance. Methods: Chemotherapeutic agents were diluted with coating buffer and coated onto 96 well plates. We then had these agents cross-react with purified clusterin and wash the wells to remove residual clusterin. We quantified the amount of clusterin with optical density (OD) measured by binding peroxidase-conjugated secondary antibody associated with mouse monoclonal clusterin antibody. To determine if anticancer drug-clusterin binding is related to chemotherapeutic agent resistance, we compared survival rates in the SKOV-3 cell line, which rarely secretes clusterin. We compared a group of SKOV-3 cells treated with a chemotherapeutic agent and a group treated with both the agent and clusterin, by means of XTT. Results: In binding tests using ELISA OD, ratios of paclitaxel, cisplatin, carboplatin, topotecan, Adriamycin, etoposide, and 5-fluoruracil (5-FU) were 2.34, 2.40, 0.52, 2.44, 1.602, 1.14, and 1.13, respectively. Topotecan, cisplatin, and paclitaxel showed relatively higher binding. In addition, when these drugs were treated with clusterin in SKOV-3 cells, anticancer resistance increased (P<0.05). Conclusions: The anticancer drug resistance endowed by clusterin is considered to be related to its binding with chemotherapeutic agents.

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