Effects of Doxorubicin, Cisplatin, and Tamoxifen on the Metabolic Profile of Human Breast Cancer MCF-7 Cells As Determined by ¹H High-Resolution Magic Angle Spinning Nuclear Magnetic Resonance

Abstract

Doxorubicin (Doxo), cisplatin (Cis), and tamoxifen (Tamo) are part of many chemotherapeutic regimens. However, there have been limited studies of the way metabolism in breast cancer is affected by chemotherapy. We studied, through H high-resolution magic angle spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy, the metabolic profile of human breast cancer MCF-7 control (Con) cells as well as MCF-7 cells treated with Tamo, Cis, and Doxo.H HR-MAS NMR single-pulse spectra evidenced signals from the cell compounds, including fatty acids (membranes), water-soluble proteins, and metabolites. The spectra showed that phosphocholine (i.e., biomarker of breast cancer malignant transformation) signals were stronger in Con than in treated cells. Betaine (i.e., the major osmolyte in cells) was observed at similar concentrations in MCF-7 control and treated cells but was absent in nontumor MCF-10A cells. The NMR spectra acquired with the Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence were used only in qualitative analyses because the signal areas were attenuated according to their transverse relaxation time (T). The CPMG method was used to identify soluble metabolites such as organic acids, amino acids, choline and its derivatives, taurine, and guanidino acetate. H HR-MAS NMR spectroscopy efficiently demonstrated the effects of Tamo, Cis, and Doxo on the metabolic profile of MCF-7 cells. The fatty acid, phosphocholine, and choline variations observed by single-pulse HR-MAS NMR have the potential to characterize both responder and nonresponder tumors at a molecular level.

Impact of chemotherapy on metabolic reprogramming: Characterization of the metabolic profile of breast cancer MDA-MB-231 cells using 1 H HR-MAS NMR spectroscopy

“…All spectra displayed in Fig. 1 showed strong, broad peaks overlapped with sharp, weak ones, the latter being assigned to water-soluble metabolites similar to those observed in MCF-7 cells [11]. The broad, strong peaks at 0.9 [1] and 1.3 ppm [2] have been assigned to methyl and methylene groups of fatty acids and proteins, respectively.…”

“…We have previously studied the influence of chemotherapy on metabolic profile of the estrogen receptor-positive human breast cancer cell line MCF-7 using 1 H high-resolution magic angle spinning (HR-MAS) NMR spectroscopy [11]. This technique confirmed that doxorubicin, cisplatin, and tamoxifen had strong effects on the metabolic profile of MCF-7 cells [11].…”

“…The vertical axes of spectra A' to D' were four-fold magnified. Numbers indicate the assignments to methyl [1] and methylene groups [2] of fatty acids and proteins, respectively; to alanine [3], acetate [4], phosphocholine [5], betaine [6], and guanidine acetate [7]; and to olefinic hydrogens of unsaturated fatty acids [8], tyrosine [9], phenylalanine [10], and UTP/UDP/UMP [11]. A to D indicate spectra from 0.5 to 4.5 ppm, whereas A' to D' indicate spectra from 5.0 to 10.0 ppm.…”

“…2 shows CPMG spectra in which sixteen metabolites have been assigned: lactate/threonine (1/2), alanine [3], acetate [4], glutamate/glutamine (5/6), acetone [7], creatine [8], choline [9], phosphocholine [10], taurine [11], glycine [12], guanidine acetate [13], tyrosine [14], phenylalanine [15], and uridinetriphosphate/uridine-diphosphate/uridine-monophosphate (UTP/UDP/UMP) [16]. These assignments have been published elsewhere [14,11]. [12], cisplatin (C and C') [13], and doxorubicin (D and D') [12].…”

“…The vertical axes of spectra A' to D' were four-fold magnified. Numbers indicate peaks attributed to lactate/threonine (1/2), alanine [3], acetate [4], glutamate/glutamine (5/6), acetone [7], creatine [8], choline [9], phosphocholine [10], taurine [11], glycine [12], guanidine acetate [13], tyrosine [14], phenylalanine [15], and UTP/UDP/UMP [16]. A to D indicate spectra from 0.5 to 4.5 ppm, whereas A' to D' indicate spectra from 5.0 to 10.0 ppm.…”