Detection of drug-induced apoptosis in human cancer cells using proton magnetic resonance (¹H-NMR) spectroscopy

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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 ELISA<sup>PLUS</sup> 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 <sup>1</sup>H-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 ELISAH<sup>PLUS</sup> assay which had reached an optimum level at the 12<sup>th</sup> hour time interval into the experiment. The maximum release of LDH was also detected at the 18<sup>th</sup> 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 ELISA<sup>PLUS</sup> assay, showing that MOLT-4 cells had likely to have undergone apoptosis from the treatment with 32 μM of cisplatin. <sup>1</sup>H-NMR spectroscopy detected a change in the ratios of methyl and methylene resonances in the MOLT-4 cells. However, the results from <sup>1</sup>H-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.