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. 1991 Feb;63(2):205–212. doi: 10.1038/bjc.1991.50

NMR spectroscopy analysis of phosphorus metabolites and the effect of adriamycin on these metabolite levels in an adriamycin-sensitive and -resistant human small cell lung carcinoma cell line.

S de Jong 1, N H Mulder 1, E G de Vries 1, G T Robillard 1
PMCID: PMC1971796  PMID: 1847646

Abstract

31P nuclear magnetic resonance (NMR) spectra of cells and of cell extract revealed high levels of phosphorylcholine (PC) and phosphocreatine (PCr) in an adriamycin-resistant human small cell lung carcinoma cell line (GLC4/ADR) and the adriamycin-sensitive parental cell line (GLC4). PCr levels in extracts of GLC4/ADR were increased compared to extracts of GLC4. We estimated that 11% of the total intracellular ATP is not bound to Mg2+ in both cell lines. This value corresponded to an intracellular free Mg2+ of 0.30 mM. The effects of different adriamycin concentrations, 0.05, 1 and 30 microM for GLC4 and 1, 30 and 200 microM for GLC4/ADR, on the phosphorus metabolite levels in continuously perfused cells were monitored. Significant differences between GLC4 and GLC4/ADR included: (a) a strong increase in the beta ATP level in the presence of 30 microM adriamycin in GLC4 only, followed by a fast decrease after 5 h of perfusion. (b) a less dramatic increase in the PC level in GLC4/ADR and an unchanged ATP level in the presence of increasing adriamycin concentrations. (c) an increased GPC level in GLC4/ADR in the presence of adriamycin. The changes in PC and GPC levels in the presence of adriamycin suggested that the phospholipid turnover was increased in GLC4/ADR and could be stimulated in the presence of adriamycin. In both cell lines, PCr levels decreased faster than the ATP levels after adriamycin treatment. Thus, biochemical markers for adriamycin resistance can be detected with NMR spectroscopy. However, more studies are necessary to obtain parameters to distinguish drug-sensitive from drug-resistant tumours in patients by NMR spectroscopy.

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Selected References

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  1. Beck W. T., Cirtain M. C., Danks M. K., Felsted R. L., Safa A. R., Wolverton J. S., Suttle D. P., Trent J. M. Pharmacological, molecular, and cytogenetic analysis of "atypical" multidrug-resistant human leukemic cells. Cancer Res. 1987 Oct 15;47(20):5455–5460. [PubMed] [Google Scholar]
  2. Bessman S. P., Carpenter C. L. The creatine-creatine phosphate energy shuttle. Annu Rev Biochem. 1985;54:831–862. doi: 10.1146/annurev.bi.54.070185.004151. [DOI] [PubMed] [Google Scholar]
  3. Cohen J. S., Lyon R. C., Chen C., Faustino P. J., Batist G., Shoemaker M., Rubalcaba E., Cowan K. H. Differences in phosphate metabolite levels in drug-sensitive and -resistant human breast cancer cell lines determined by 31P magnetic resonance spectroscopy. Cancer Res. 1986 Aug;46(8):4087–4090. [PubMed] [Google Scholar]
  4. Daly P. F., Cohen J. S. Magnetic resonance spectroscopy of tumors and potential in vivo clinical applications: a review. Cancer Res. 1989 Feb 15;49(4):770–779. [PubMed] [Google Scholar]
  5. Daly P. F., Lyon R. C., Faustino P. J., Cohen J. S. Phospholipid metabolism in cancer cells monitored by 31P NMR spectroscopy. J Biol Chem. 1987 Nov 5;262(31):14875–14878. [PubMed] [Google Scholar]
  6. Desmoulin F., Galons J. P., Canioni P., Marvaldi J., Cozzone P. J. 31P nuclear magnetic resonance study of a human colon adenocarcinoma cultured cell line. Cancer Res. 1986 Aug;46(8):3768–3774. [PubMed] [Google Scholar]
  7. Estey E. H., Silberman L., Beran M., Andersson B. S., Zwelling L. A. The interaction between nuclear topoisomerase II activity from human leukemia cells, exogenous DNA, and 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA) or 4-(4,6-O-ethylidene-beta-D-glucopyranoside) (VP-16) indicates the sensitivity of the cells to the drugs. Biochem Biophys Res Commun. 1987 Apr 29;144(2):787–793. doi: 10.1016/s0006-291x(87)80033-5. [DOI] [PubMed] [Google Scholar]
  8. Evanochko W. T., Ng T. C., Lilly M. B., Lawson A. J., Corbett T. H., Durant J. R., Glickson J. D. In vivo 31P NMR study of the metabolism of murine mammary 16/C adenocarcinoma and its response to chemotherapy, x-radiation, and hyperthermia. Proc Natl Acad Sci U S A. 1983 Jan;80(2):334–338. doi: 10.1073/pnas.80.2.334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evanochko W. T., Sakai T. T., Ng T. C., Krishna N. R., Kim H. D., Zeidler R. B., Ghanta V. K., Brockman R. W., Schiffer L. M., Braunschweiger P. G. NMR study of in vivo RIF-1 tumors. Analysis of perchloric acid extracts and identification of 1H, 31P and 13C resonances. Biochim Biophys Acta. 1984 Sep 14;805(1):104–116. doi: 10.1016/0167-4889(84)90042-9. [DOI] [PubMed] [Google Scholar]
  10. Evans F. E., Kaplan N. O. 31P nuclear magnetic resonance studies of HeLa cells. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4909–4913. doi: 10.1073/pnas.74.11.4909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Evelhoch J. L., Keller N. A., Corbett T. H. Response-specific adriamycin sensitivity markers provided by in vivo 31P nuclear magnetic resonance spectroscopy in murine mammary adenocarcinomas. Cancer Res. 1987 Jul 1;47(13):3396–3401. [PubMed] [Google Scholar]
  12. Ferguson P. J., Fisher M. H., Stephenson J., Li D. H., Zhou B. S., Cheng Y. C. Combined modalities of resistance in etoposide-resistant human KB cell lines. Cancer Res. 1988 Nov 1;48(21):5956–5964. [PubMed] [Google Scholar]
  13. Glisson B., Gupta R., Smallwood-Kentro S., Ross W. Characterization of acquired epipodophyllotoxin resistance in a Chinese hamster ovary cell line: loss of drug-stimulated DNA cleavage activity. Cancer Res. 1986 Apr;46(4 Pt 2):1934–1938. [PubMed] [Google Scholar]
  14. Gupta R. K., Moore R. D. 31P NMR studies of intracellular free Mg2+ in intact frog skeletal muscle. J Biol Chem. 1980 May 10;255(9):3987–3993. [PubMed] [Google Scholar]
  15. Gupta R. K., Yushok W. D. Noninvasive 31P NMR probes of free Mg2+, MgATP, and MgADP in intact Ehrlich ascites tumor cells. Proc Natl Acad Sci U S A. 1980 May;77(5):2487–2491. doi: 10.1073/pnas.77.5.2487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Guy G. R., Murray A. W. Tumor promoter stimulation of phosphatidylcholine turnover in HeLa cells. Cancer Res. 1982 May;42(5):1980–1985. [PubMed] [Google Scholar]
  17. Horio M., Gottesman M. M., Pastan I. ATP-dependent transport of vinblastine in vesicles from human multidrug-resistant cells. Proc Natl Acad Sci U S A. 1988 May;85(10):3580–3584. doi: 10.1073/pnas.85.10.3580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Knop R. H., Carney D. N., Chen C. W., Cohen J. S., Minna J. D. Levels of high energy phosphates in human lung cancer cell lines by 31P nuclear magnetic resonance spectroscopy. Cancer Res. 1987 Jul 1;47(13):3357–3359. [PubMed] [Google Scholar]
  19. Knop R. H., Chen C. W., Mitchell J. B., Russo A., McPherson S., Cohen J. S. Metabolic studies of mammalian cells by 31P-NMR using a continuous perfusion technique. Biochim Biophys Acta. 1984 Jul 20;804(3):275–284. doi: 10.1016/0167-4889(84)90130-7. [DOI] [PubMed] [Google Scholar]
  20. Lawson J. W., Veech R. L. Effects of pH and free Mg2+ on the Keq of the creatine kinase reaction and other phosphate hydrolyses and phosphate transfer reactions. J Biol Chem. 1979 Jul 25;254(14):6528–6537. [PubMed] [Google Scholar]
  21. Li S. J., Wehrle J. P., Rajan S. S., Steen R. G., Glickson J. D., Hilton J. Response of radiation-induced fibrosarcoma-1 in mice to cyclophosphamide monitored by in vivo 31P nuclear magnetic resonance spectroscopy. Cancer Res. 1988 Sep 1;48(17):4736–4742. [PubMed] [Google Scholar]
  22. Lyon R. C., Cohen J. S., Faustino P. J., Megnin F., Myers C. E. Glucose metabolism in drug-sensitive and drug-resistant human breast cancer cells monitored by magnetic resonance spectroscopy. Cancer Res. 1988 Feb 15;48(4):870–877. [PubMed] [Google Scholar]
  23. Meijer C., Mulder N. H., Timmer-Bosscha H., Zijlstra J. G., de Vries E. G. Role of free radicals in an adriamycin-resistant human small cell lung cancer cell line. Cancer Res. 1987 Sep 1;47(17):4613–4617. [PubMed] [Google Scholar]
  24. Morash S. C., Cook H. W., Spence M. W. Phosphatidylcholine metabolism in cultured cells: catabolism via glycerophosphocholine. Biochim Biophys Acta. 1988 Jul 22;961(2):194–202. doi: 10.1016/0005-2760(88)90114-2. [DOI] [PubMed] [Google Scholar]
  25. Naruse S., Hirakawa K., Horikawa Y., Tanaka C., Higuchi T., Ueda S., Nishikawa H., Watari H. Measurements of in vivo 31P nuclear magnetic resonance spectra in neuroectodermal tumors for the evaluation of the effects of chemotherapy. Cancer Res. 1985 Jun;45(6):2429–2433. [PubMed] [Google Scholar]
  26. Pastan I., Gottesman M. Multiple-drug resistance in human cancer. N Engl J Med. 1987 May 28;316(22):1388–1393. doi: 10.1056/NEJM198705283162207. [DOI] [PubMed] [Google Scholar]
  27. Pommier Y., Kerrigan D., Schwartz R. E., Swack J. A., McCurdy A. Altered DNA topoisomerase II activity in Chinese hamster cells resistant to topoisomerase II inhibitors. Cancer Res. 1986 Jun;46(6):3075–3081. [PubMed] [Google Scholar]
  28. Riordan J. R., Ling V. Genetic and biochemical characterization of multidrug resistance. Pharmacol Ther. 1985;28(1):51–75. doi: 10.1016/0163-7258(85)90082-8. [DOI] [PubMed] [Google Scholar]
  29. Sinha B. K., Katki A. G., Batist G., Cowan K. H., Myers C. E. Differential formation of hydroxyl radicals by adriamycin in sensitive and resistant MCF-7 human breast tumor cells: implications for the mechanism of action. Biochemistry. 1987 Jun 30;26(13):3776–3781. doi: 10.1021/bi00387a006. [DOI] [PubMed] [Google Scholar]
  30. Steen R. G. Response of solid tumors to chemotherapy monitored by in vivo 31P nuclear magnetic resonance spectroscopy: a review. Cancer Res. 1989 Aug 1;49(15):4075–4085. [PubMed] [Google Scholar]
  31. Steen R. G., Tamargo R. J., McGovern K. A., Rajan S. S., Brem H., Wehrle J. P., Glickson J. D. In vivo 31P nuclear magnetic resonance spectroscopy of subcutaneous 9L gliosarcoma: effects of tumor growth and treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea on tumor bioenergetics and histology. Cancer Res. 1988 Feb 1;48(3):676–681. [PubMed] [Google Scholar]
  32. Yeh G. C., Occhipinti S. J., Cowan K. H., Chabner B. A., Myers C. E. Adriamycin resistance in human tumor cells associated with marked alteration in the regulation of the hexose monophosphate shunt and its response to oxidant stress. Cancer Res. 1987 Nov 15;47(22):5994–5999. [PubMed] [Google Scholar]
  33. Zijlstra J. G., de Vries E. G., Mulder N. H. Multifactorial drug resistance in an adriamycin-resistant human small cell lung carcinoma cell line. Cancer Res. 1987 Apr 1;47(7):1780–1784. [PubMed] [Google Scholar]
  34. Zijlstra J. G., de Vries E. G., Muskiet F. A., Martini I. A., Timmer-Bosscha H., Mulder N. H. Influence of docosahexaenoic acid in vitro on intracellular adriamycin concentration in lymphocytes and human adriamycin-sensitive and -resistant small-cell lung cancer cell lines, and on cytotoxicity in the tumor cell lines. Int J Cancer. 1987 Dec 15;40(6):850–856. doi: 10.1002/ijc.2910400625. [DOI] [PubMed] [Google Scholar]
  35. de Jong S., Zijlstra J. G., de Vries E. G., Mulder N. H. Reduced DNA topoisomerase II activity and drug-induced DNA cleavage activity in an adriamycin-resistant human small cell lung carcinoma cell line. Cancer Res. 1990 Jan 15;50(2):304–309. [PubMed] [Google Scholar]

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