NMR spectroscopy-based metabolomics of human hepatocarcinoma cell line:
an integrated approach
to tumor metabolome.
F. Conti
In analogy to the expressions “tumor genome” and “tumor proteome”, Mazurek S. and Eigenbrodt E. referred “tumor metabolome” as the peculiar metabolic characteristics of tumor cells (Mazurek and Eigenbrodt Anticancer Res 2003). The tumor metabolome is characterized by high glycolytic and glutaminolytic capacities and high channeling of glucose carbons to synthetic processes. This metabolic phenotype is, however, expressed in the context of the microenvironment, like substrate or favouring growth factor availability, which profoundly determines the enzyme activities and substrate utilization, as well as the adaptative rearrangement within and among metabolic pathways. NMR-based metabolomics shows to be a particularly sensitive method to monitor changes in a biological system, through the integrated analysis of variations in the metabolic network.
In the present communication we report a NMR-based metabolomic approach to study the adaptative changes in the metabolome of human hepatocarcinoma cells (HepG2) to the density variations of cell cultures. It is well established that under in vitro conditions the cell density influences both cell physiology and cellular response to antitumor drugs, even if cell growth is not completely inhibited in HepG2 cell cultures. On these bases, we studied the HepG2 metabolome by using 1H NMR spectroscopy and multivariate data analysis on cell culture medium before and after 24 hours of culture.
A. Miccheli*, A. Tomassini*, R. Di Clemente*, M. Valerio*, G. Capuani*, M. Bizzarri**, L.Conti Devirgiliis***, F. Conti*. * Department of Chemistry, “La Sapienza” University of Rome, Italy
** Department of Experimental Medicine, “La Sapienza” University of Rome,
*** University of L’Aquila, Italy
13C nmr
spectroscopy-based
metabolic profiling
of cancer cell growth.
C. Puccetti
Metabolic profiling is defined as the simultaneous assessment of substrate flux within and among major pathways of macromolecule and molecules synthesis and energy production under various physiological conditions, growth phases and substrate availability.
In the last years, the metabolic profiling has been performed by using mass spectroscopy (MS) and [13C] labeled substrates. The distribution of labeled carbons in various intermediates during the de novo macromolecule synthesis in cancer cells has been revealed allowing to define the role of several metabolic processes in cancer cell growth and death (Boros L.G., Cascante M. and Lee W-N.L DDT 2002; Lee W-N.L. et al. Am J Physiol 1998). In this comunication we report the metabolic profiling and isotopomer analysis of Jurkat cells as obtained by 13C NMR spectroscopy using [1,2-13C2]glucose. The obtained results reveal an heterogeneity of metabolism in an asyncronous cell population that is interpreted on the basis of different metabolic phenotypes of subpopulations due to the presence of cells in different cycle phases.
Metabolic profiling by using stable-isotope tracer technology is showing to be a powerfull integrated investigative tool to reveal the tumor-specific metabolic shifts and to identify the metabolic steps controlling cell proliferation, to be used also for the identification of new anti-ca ncer targets.
A. Miccheli*, C. Puccetti*,
A. Tomassini*, M. Valerio*, G. Peluso**, F. Tuccillo***,
M. Calvani****, F. Conti*. * Department of Chemistry, “La Sapienza” University of Rome, Italy
** Institute of Biochemistry of Proteins, CNR, Naples, Italy,
*** National Cancer Institute-INT “Fondazione G. Pascale,” Naples, Italy,
**** Scientific Department, Sigma Tau, Rome, Italy.
In analogy to the expressions “tumor genome” and “tumor proteome”, Mazurek S. and Eigenbrodt E. referred “tumor metabolome” as the peculiar metabolic characteristics of tumor cells (Mazurek and Eigenbrodt Anticancer Res 2003). The tumor metabolome is characterized by high glycolytic and glutaminolytic capacities and high channeling of glucose carbons to synthetic processes. This metabolic phenotype is, however, expressed in the context of the microenvironment, like substrate or favouring growth factor availability, which profoundly determines the enzyme activities and substrate utilization, as well as the adaptative rearrangement within and among metabolic pathways.
NMR-based metabolomics shows to be a particularly sensitive method to monitor changes in a biological system, through the integrated analysis of variations in the metabolic network.
Metabolic profiling is defined as the simultaneous assessment of substrate flux within and among major pathways of macromolecule and molecules synthesis and energy production under various physiological conditions, growth phases and substrate availability. 
In this comunication we report the metabolic profiling and isotopomer analysis of Jurkat cells as obtained by 13C NMR spectroscopy using [1,2-13C2]glucose. The obtained results reveal an heterogeneity of metabolism in an asyncronous cell population that is interpreted on the basis of different metabolic phenotypes of subpopulations due to the presence of cells in different cycle phases.