Korean J Obstet Gynecol.  2008 Jul;51(7):738-743.

The correlation of clusterin binding affinity to chemotherapeutic agents with chemoresistance in ovarian cancer cells

• Choi JH
• Suh MJ
• Park DC

Abstract

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.

Keyword

Clusterin; Drug binding; Chemo-resistance; Ovarian cancer

MeSH Terms

Animals
Carboplatin
Cell Line
Cisplatin
Clusterin
Doxorubicin
Drug Resistance
Enzyme-Linked Immunosorbent Assay
Etoposide
Mice
Ovarian Neoplasms
Paclitaxel
Survival Rate
Topotecan
Carboplatin
Cisplatin
Clusterin
Doxorubicin
Etoposide
Paclitaxel
Topotecan