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Detection of drug-induced apoptosis in human cancer cells using proton magnetic resonance (¹H-NMR) spectroscopy
Author
Choo, Kelvin Kuen Yew
Supervisor
Lee, Peter Peng Foo
Yan, Yaw Kai
Abstract
Cisplatin is a well-known and established anticancer drug [1]. In this study, MOLT-4, a human leukemia cell line, was treated with 32 μM cisplatin after which various cell death assays were carried out to determine the presence of nucleosomes using the ELISAPLUS assay and the level of Lactate dehydrogenase (LDH) activity. The objectives in this study were to examine the cell death processes that were occurring in this treatment and to determine if 1H-NMR spectroscopy could be used as a viable alternative technique to measure cell death, e.g. apoptosis and/or necrosis.
Results obtained from the cell death assays showed that cisplatin induced apoptosis on MOLT-4 cells. This was shown by the release of nucleosomes using the ELISAHPLUS assay which had reached an optimum level at the 12th hour time interval into the experiment. The maximum release of LDH was also detected at the 18th hour into the experiment. The release of LDH was due to the loss of the structural integrity of the MOLT-4 cells’ plasma membrane. This phenomenon had supported the results from the release of nucleosomes using the ELISAPLUS assay, showing that MOLT-4 cells had likely to have undergone apoptosis from the treatment with 32 μM of cisplatin. 1H-NMR spectroscopy detected a change in the ratios of methyl and methylene resonances in the MOLT-4 cells. However, the results from 1H-NMR spectroscopy had also indicated the presence of late apoptosis and/or necrosis. Hence, this technique may not be useful to differentiate between apoptosis and necrosis.
Results obtained from the cell death assays showed that cisplatin induced apoptosis on MOLT-4 cells. This was shown by the release of nucleosomes using the ELISAHPLUS assay which had reached an optimum level at the 12th hour time interval into the experiment. The maximum release of LDH was also detected at the 18th hour into the experiment. The release of LDH was due to the loss of the structural integrity of the MOLT-4 cells’ plasma membrane. This phenomenon had supported the results from the release of nucleosomes using the ELISAPLUS assay, showing that MOLT-4 cells had likely to have undergone apoptosis from the treatment with 32 μM of cisplatin. 1H-NMR spectroscopy detected a change in the ratios of methyl and methylene resonances in the MOLT-4 cells. However, the results from 1H-NMR spectroscopy had also indicated the presence of late apoptosis and/or necrosis. Hence, this technique may not be useful to differentiate between apoptosis and necrosis.
Date Issued
2009
Call Number
QD96.P7 Cho
Date Submitted
2009