Detection of drug-induced apoptosis and necrosis in human cervical carcinoma cells using 1H NMR spectroscopy

Abstract

Apoptosis and necrosis need to be differentiated in order to distinguish drug-induced cell death from spontaneous cell death due to hypoxia. The ability to differentiate between these two modes of cell death, especially at an early stage in the process, could have a significant impact on accessing the outcome of anticancer drug therapy in the clinic. Nuclear magnetic resonance spectroscopy was used to distinguish apoptosis from necrosis in human cervical carcinoma (HeLa) cells. Apoptosis was induced by treatment with the topoisomerase II inhibitor etoposide, whereas necrosis was induced by the use of ethacrynic acid or cytochalasin B. We found that the intensity of the methylene resonance increases significantly as early as 6 h after the onset of apoptosis, but that no such changes occur during necrosis. The spectral intensity ratio of the methylene to methyl resonances also shows a high correlation with the percentage of apoptotic cells in the sample (r 2 =0.965, P50.003). Cell Death and Differentiation (2001) 8, 219 ± 224.

Noninvasive estimation of tumour viability in a xenograft model of human neuroblastoma with proton magnetic resonance spectroscopy (1H MRS)

“…Furthermore, in vitro studies have shown that the ratio of the methylene (CH 2 ) lipid resonance (at 1.3 ppm) to the methyl (CH 3 ) resonance (at 0.9 ppm) is correlated to the percentage of apoptotic lymphoblasts after doxorubicin treatment (Blankenberg et al, 1997). This finding was recently confirmed using a human cervical carcinoma HeLa cell system (Bezabeh et al, 2001). In the present study, we found strong ML signals in large, highly necrotic tumours.…”

EBNA3C-Mediated Regulation of Aurora Kinase B Contributes to Epstein-Barr Virus-Induced B-Cell Proliferation through Modulation of the Activities of the Retinoblastoma Protein and Apoptotic Caspases

“…Similarly, various reports have suggested that deregulation of AK-B by either knockdown or disruption of the functional kinase domain leads to apoptosis (26,36,69,70). This led us to propose that AK-B combined with EBNA3C would promote resistance to apoptosis.…”

N-(4-iodophenyl)-N′-(2-chloroethyl)urea as a microtubule disrupter: in vitro and in vivo profiling of antitumoral activity on CT-26 murine colon carcinoma cell line cultured and grafted to mice

“…The early lipid signal changes probably arose from the interactions described recently and occurring between ICEU and the lipid bilayer membranes (Saint-Laurent et al, 2001). The late lipid signal changes (observed after 18 h) may be considered as markers of the apoptotic cascade, therefore reflecting plasma membrane reorganisation and phosphatidylserine externalisation (Blankenberg et al, 1996(Blankenberg et al, , 1997Bezabeh et al, 2001;Brisdelli et al, 2003). Moreover, after the latter stage, there was a decrease in choline that indicates cell death (Blankenberg et al, 1996(Blankenberg et al, , 1997.…”

“…For data analysis, each spectrum was phased and baseline-corrected using XWIN-NMR version 2.6 (Bruker software). The resonances were assigned based on the known chemical shifts of the major structural groups and metabolites using published data (Bezabeh et al, 2001;Sitter et al, 2002;Brisdelli et al, 2003) and commercially available pure metabolite reference spectra. The relative content of metabolites was estimated by peak area integration using PeakFit software (version 4, SPSS Science, Chicago, IL, USA).…”