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- Title
<sup>1</sup>H NMR and hyperpolarized <sup>13</sup>C NMR assays of pyruvate-lactate: a comparative study.
- Authors
Hill, Deborah K.; Jamin, Yann; Orton, Matthew R.; Tardif, Nicolas; Parkes, Harold G.; Robinson, Simon P.; Leach, Martin O.; Chung, Yuen ‐ Li; Eykyn, Thomas R.
- Abstract
Pyruvate-lactate exchange is mediated by the enzyme lactate dehydrogenase (LDH) and is central to the altered energy metabolism in cancer cells. The measurement of exchange kinetics using hyperpolarized 13C NMR has provided a biomarker of response to novel therapeutics. However, the observable signal is restricted to the exchanging hyperpolarized 13C pools and the endogenous pools of 12C-labelled metabolites are invisible in these measurements. In this study, we investigated an alternative in vitro 1H NMR assay, using [3-13C]pyruvate, and compared the measured kinetics with a hyperpolarized 13C NMR assay, using [1-13C]pyruvate, under the same conditions in human colorectal carcinoma SW1222 cells. The apparent forward reaction rate constants ( kPL) derived from the two assays showed no significant difference, and both assays had similar reproducibility ( kPL = 0.506 ± 0.054 and kPL = 0.441 ± 0.090 nmol/s/106 cells; mean ± standard deviation; n = 3); 1H, 13C assays, respectively). The apparent backward reaction rate constant ( kLP) could only be measured with good reproducibility using the 1H NMR assay ( kLP = 0.376 ± 0.091 nmol/s/106 cells; mean ± standard deviation; n = 3). The 1H NMR assay has adequate sensitivity to measure real-time pyruvate-lactate exchange kinetics in vitro, offering a complementary and accessible assay of apparent LDH activity. Copyright © 2013 John Wiley & Sons, Ltd.
- Publication
NMR in Biomedicine, 2013, Vol 26, Issue 10, p1321
- ISSN
0952-3480
- Publication type
Academic Journal
- DOI
10.1002/nbm.2957