Detection of polyol accumulation in a new ovarian carcinoma cell line, CABA I: a1H NMR study

Abstract

Ovarian carcinomas represent a major form of gynaecological malignancies, whose treatment consists mainly of surgery and chemotherapy. Besides the difficulty of prognosis, therapy of ovarian carcinomas has reached scarce improvement, as a consequence of lack of efficacy and development of drug-resistance. The need of different biochemical and functional parameters has grown, in order to obtain a larger view on processes of biological and clinical significance. In this paper we report novel metabolic features detected in a series of different human ovary carcinoma lines, by 1H NMR spectroscopy of intact cells and their extracts. Most importantly, a new ovarian adenocarcinoma line CABA I, showed strong signals in the spectral region between 3.5 and 4.0 p.p.m., assigned for the first time to the polyol sorbitol (39±11 nmol/106 cells). 13C NMR analyses of these cells incubated with [1-13C]-D-glucose demonstrated labelled-sorbitol formation. The other ovarian carcinoma cell lines (OVCAR-3, IGROV 1, SK-OV-3 and OVCA432), showed, in the same spectral region, intense resonances from other metabolites: glutathione (up to 30 nmol/106 cells) and myo-inositol (up to 50 nmol/106 cells). Biochemical and biological functions are suggested for these compounds in human ovarian carcinoma cells, especially in relation to their possible role in cell detoxification mechanisms during tumour progression.British Journal of Cancer (2002) 86, 1180–1187. DOI: 10.1038/sj/bjc/6600189 www.bjcancer.com© 2002 Cancer Research UK

Determination of thiols and disulfides via HPLC quantification of 5-thio-2-nitrobenzoic acid

“…Although no reports are available on the quantities of non-protein thiols, protein thiols, non-protein disulfides, and protein disulfides in OVCAR-3 cells, the quantity of GSH determined to be 12.16±0.05 nmol/million cells before reduction in non-protein thiol samples by this method (Table 1) is in agreement with the literature reported values for the cells, which are between 7.9±1.3 nmol/million cells [34] and 29.5 nmol/million cells [35]. …”

Alterations of Choline Phospholipid Metabolism in Ovarian Tumor Progression

“…Cells were washed twice with ice-cold physiologic saline solution and pellets resuspended in 0.5 mL ice-cold twice-distilled water. Aqueous extracts (20 Â 10 6 -30 Â 10 6 cells/sample) were prepared in ethanol/H 2 O (70:30, v/v) according to an established protocol already described (17,24,25). This method was proven to be as efficient as the more commonly used extraction in perchloric acid, with the additional advantages of higher procedural simplicity (26,27).…”

“…High-resolution NMR experiments (25jC) were done at either 400 or 700 MHz (Bruker AVANCE spectrometers, Karlsruhe, Germany). 1 H NMR spectra of cell extracts were obtained using acquisition pulses, water presaturation, data processing, and data analysis as described already (17). Quantification of individual metabolites was obtained from peak areas using correction factors determined by experiments at the equilibrium of magnetization (90j pulses, 30.00-second interpulse delay).…”

“…To date, choline phospholipid metabolism in ovarian cancer has received only limited attention despite preliminary indications of increased total choline in human ovary tumors and biopsies (15,16) and accumulation of phosphocholine in ovarian carcinoma cell lines (17). In the present study, we quantified the intracellular levels of phosphatidylcholine metabolites in the progression from nontumoral ovarian surface epithelial (OSE) cells or immortalized cell variants to ovarian cancer cell lines and related these levels to quantitative differences in basal activity of enzymes involved in phosphatidylcholine biosynthesis and/or catabolism (see Fig.…”

Metabolic Markers of MG-63 Osteosarcoma Cell Line Response to Doxorubicin and Methotrexate Treatment: Comparison to Cisplatin

“…Depletion in minositol has been previously detected in several studies, 27,46,53 and such variation was assigned to several possible causes: disturbance in its osmolytic function through drug-induced partial inhibition of the Na + /m-inositol cotransporter (SMIT); 46 diminished inhibition of the mitogen-activated protein kinase (MAPK) signal transduction pathway, thus leading to stimulation of cells into apoptosis; 46 effect on the biosynthesis and turnover of some phospholipids; 54 or activation of cell detoxification processes. To clarify the role of inositols in treated MG-63 cells (particularly upon cDDP and DOX treatments), STOCSY analysis of the inositol resonances (not shown) revealed that, in cDDP-treated cells, m-inositol decreases concomitantly with Tau, Glu/Gln, and creatine (besides sinositol), whereas in DOX-treated cells, m-inositol decreases together with Glu/Gln and GSH (and s-inositol).…”