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KMID : 0358420100530010043
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Korean Journal of Obstetrics and Gynecology
2010 Volume.53 No. 1 p.43 ~ p.52
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Detection of chemosensitivity using K18-Asp396 (M30) antibody in HeLa and OVCAR-3 cell lines treated with anticancer agents
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Song Min-Kyung
Kwack Hyun-Sung
Ryu Ki-Sung
Han Ku-Taek
Park Sang-Ho
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Abstract
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Objective: The aim of this study was to detect the levels of M30-antigens as a biomarker of apoptosis in cells and their culture media after treatments with anticancer drugs as a preclinical study.
Methods: After HeLa and OVCAR-3 cells were treated respectively with paclitaxel, cisplatin, and camptothecin, the harvested cells were stained sequentially with M30 monoclonal antibodies and propidium iodide (PI). Afterwards, they were analyzed using a FACScan flow cytometer and observed under an immunofluorescence microscope for M30-FITC immunofluorescences. Levels of M30 antigens were also detected in their culture media using M30-Apoptosense ELISA kit.
Results: The levels of M30-FITC immunofluorescences were elevated in both cell lines after each drug treatments compared with those of control cells. The levels of M30 antigens detected by ELISA in media culturing each cell line treated with each of drugs were elevated compared with those of control cells.
Conclusion: This study suggests that M30-antigens representing chemotherapy induced apoptosis may be a useful biomarker for predicting and monitoring the response of neoadjuvant chemotherapy in patients with gynecologic cancers.
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KEYWORD
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Apoptosis, Cytokeratin 18-Asp396 (M30), Flow cytometry, ELISA, HeLa, OVCAR-3
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