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. 1993 Dec;68(6):1061–1070. doi: 10.1038/bjc.1993.483

31P-nuclear magnetic resonance spectroscopy in vivo of six human melanoma xenograft lines: tumour bioenergetic status and blood supply.

H Lyng 1, D R Olsen 1, T E Southon 1, E K Rofstad 1
PMCID: PMC1968640  PMID: 8260356

Abstract

Six human melanoma xenograft lines grown s.c. in BALB/c-nu/nu mice were subjected to 31P-nuclear magnetic resonance (31P-NMR) spectroscopy in vivo. The following resonances were detected: phosphomonoesters (PME), inorganic phosphate (Pi), phosphodiesters (PDE), phosphocreatine (PCr) and nucleoside triphosphate gamma, alpha and beta (NTP gamma, alpha and beta). The main purpose of the work was to search for possible relationships between 31P-NMR resonance ratios and tumour pH on the one hand and blood supply per viable tumour cell on the other. The latter parameter was measured by using the 86Rb uptake method. Tumour bioenergetic status [the (PCr + NTP beta)/Pi resonance ratio], tumour pH and blood supply per viable tumour cell decreased with increasing tumour volume for five of the six xenograft lines. The decrease in tumour bioenergetic status was due to a decrease in the (PCr + NTP beta)/total resonance ratio as well as an increase in the Pi/total resonance ratio. The decrease in the (PCr + NTP beta)/total resonance ratio was mainly a consequence of a decrease in the PCr/total resonance ratio for two lines and mainly a consequence of a decrease in the NTP beta/total resonance ratio for three lines. The magnitude of the decrease in the (PCr + NTP beta)/total resonance ratio and the magnitude of the decrease in tumour pH were correlated to the magnitude of the decrease in blood supply per viable tumour cell. Tumour pH decreased with decreasing tumour bioenergetic status, and the magnitude of this decrease was larger for the tumour lines showing a high than for those showing a low blood supply per viable tumour cell. No correlations across the tumour lines were found between tumour pH and tumour bioenergetic status or any other resonance ratio on the one hand and blood supply per viable tumour cell on the other. The differences in the 31P-NMR spectrum between the tumour lines were probably caused by differences in the intrinsic biochemical properties of the tumour cells rather than by the differences in blood supply per viable tumour cell. Biochemical properties of particular importance included rate of respiration, glycolytic capacity and tolerance to hypoxic stress. On the other hand, tumour bioenergetic status and tumour pH were correlated to blood supply per viable tumour cell within individual tumour lines. These observations suggest that 31P-NMR spectroscopy may be developed to be a clinically useful method for monitoring tumour blood supply and parameters related to tumour blood supply during and after physiological intervention and tumour treatment. However, clinically useful parameters for prediction of tumour treatment resistance caused by insufficient blood supply can probably not be derived from a single 31P-NMR spectrum since correlations across tumour lines were not detected; additional information is needed.

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Selected References

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  1. Bremner J. C., Counsell C. J., Adams G. E., Stratford I. J., Wood P. J., Dunn J. F., Radda G. K. In vivo 31P nuclear magnetic resonance spectroscopy of experimental murine tumours and human tumour xenografts: effects of blood flow modification. Br J Cancer. 1991 Nov;64(5):862–866. doi: 10.1038/bjc.1991.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ceckler T. L., Bryant R. G., Penney D. P., Gibson S. L., Hilf R. 31P-NMR spectroscopy demonstrates decreased ATP levels in vivo as an early response to photodynamic therapy. Biochem Biophys Res Commun. 1986 Oct 15;140(1):273–279. doi: 10.1016/0006-291x(86)91086-7. [DOI] [PubMed] [Google Scholar]
  3. Chapman J. D., McPhee M. S., Walz N., Chetner M. P., Stobbe C. C., Soderlind K., Arnfield M., Meeker B. E., Trimble L., Allen P. S. Nuclear magnetic resonance spectroscopy and sensitizer-adduct measurements of photodynamic therapy-induced ischemia in solid tumors. J Natl Cancer Inst. 1991 Nov 20;83(22):1650–1659. doi: 10.1093/jnci/83.22.1650. [DOI] [PubMed] [Google Scholar]
  4. Corbett R. J., Nunnally R. L., Giovanella B. C., Antich P. P. Characterization of the 31P nuclear magnetic resonance spectrum from human melanoma tumors implanted in nude mice. Cancer Res. 1987 Oct 1;47(19):5065–5069. [PubMed] [Google Scholar]
  5. 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]
  6. Evanochko W. T., Ng T. C., Glickson J. D. Application of in vivo NMR spectroscopy to cancer. Magn Reson Med. 1984 Dec;1(4):508–534. doi: 10.1002/mrm.1910010410. [DOI] [PubMed] [Google Scholar]
  7. Evanochko W. T., Ng T. C., Glickson J. D., Durant J. R., Corbett T. H. Human tumors as examined by in vivo 31P NMR in athymic mice. Biochem Biophys Res Commun. 1982 Dec 31;109(4):1346–1352. doi: 10.1016/0006-291x(82)91925-8. [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. Evelhoch J. L., Sapareto S. A., Nussbaum G. H., Ackerman J. J. Correlations between 31P NMR spectroscopy and 15O perfusion measurements in the RIF-1 murine tumor in vivo. Radiat Res. 1986 Apr;106(1):122–131. [PubMed] [Google Scholar]
  11. Freyer J. P., Schor P. L., Jarrett K. A., Neeman M., Sillerud L. O. Cellular energetics measured by phosphorous nuclear magnetic resonance spectroscopy are not correlated with chronic nutrient deficiency in multicellular tumor spheroids. Cancer Res. 1991 Aug 1;51(15):3831–3837. [PubMed] [Google Scholar]
  12. Godfrey K. Statistics in practice. Comparing the means of several groups. N Engl J Med. 1985 Dec 5;313(23):1450–1456. doi: 10.1056/NEJM198512053132305. [DOI] [PubMed] [Google Scholar]
  13. Irving M. G., Simpson S. J., Field J., Doddrell D. M. Use of high-resolution 31P-labeled topical magnetic resonance spectroscopy to monitor in vivo tumor metabolism in rats. Cancer Res. 1985 Feb;45(2):481–486. [PubMed] [Google Scholar]
  14. Kallinowski F., Schlenger K. H., Runkel S., Kloes M., Stohrer M., Okunieff P., Vaupel P. Blood flow, metabolism, cellular microenvironment, and growth rate of human tumor xenografts. Cancer Res. 1989 Jul 15;49(14):3759–3764. [PubMed] [Google Scholar]
  15. Kallinowski F., Vaupel P., Runkel S., Berg G., Fortmeyer H. P., Baessler K. H., Wagner K., Mueller-Klieser W., Walenta S. Glucose uptake, lactate release, ketone body turnover, metabolic micromilieu, and pH distributions in human breast cancer xenografts in nude rats. Cancer Res. 1988 Dec 15;48(24 Pt 1):7264–7272. [PubMed] [Google Scholar]
  16. Kalra R., Wade K. E., Hands L., Styles P., Camplejohn R., Greenall M., Adams G. E., Harris A. L., Radda G. K. Phosphomonoester is associated with proliferation in human breast cancer: a 31P MRS study. Br J Cancer. 1993 May;67(5):1145–1153. doi: 10.1038/bjc.1993.211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kluge M., Elger B., Engel T., Schaefer C., Seega J., Vaupel P. Acute effects of tumor necrosis factor alpha or lymphotoxin on global blood flow, laser Doppler flux, and bioenergetic status of subcutaneous rodent tumors. Cancer Res. 1992 Apr 15;52(8):2167–2173. [PubMed] [Google Scholar]
  18. Koutcher J. A., Alfieri A. A., Barnett D. C., Cowburn D. C., Kornblith A. B., Kim J. H. Changes in 31P nuclear magnetic resonance with tumor growth in radioresistant and radiosensitive tumors. Radiat Res. 1990 Mar;121(3):312–319. [PubMed] [Google Scholar]
  19. Koutcher J. A., Alfieri A. A., Devitt M. L., Rhee J. G., Kornblith A. B., Mahmood U., Merchant T. E., Cowburn D. Quantitative changes in tumor metabolism, partial pressure of oxygen, and radiobiological oxygenation status postradiation. Cancer Res. 1992 Sep 1;52(17):4620–4627. [PubMed] [Google Scholar]
  20. Lilly M. B., Ng T. C., Evanochko W. T., Katholi C. R., Kumar N. G., Elgavish G. A., Durant J. R., Hiramoto R., Ghanta V., Glickson J. D. Loss of high-energy phosphate following hyperthermia demonstrated by in vivo 31P-nuclear magnetic resonance spectroscopy. Cancer Res. 1984 Feb;44(2):633–638. [PubMed] [Google Scholar]
  21. Lowry M., Porter D. A., Twelves C. J., Heasley P. E., Smith M. A., Richards M. A. Visibility of phospholipids in 31P NMR spectra of human breast tumours in vivo. NMR Biomed. 1992 Jan-Feb;5(1):37–42. doi: 10.1002/nbm.1940050107. [DOI] [PubMed] [Google Scholar]
  22. Lyng H., Skretting A., Rofstad E. K. Blood flow in six human melanoma xenograft lines with different growth characteristics. Cancer Res. 1992 Feb 1;52(3):584–592. [PubMed] [Google Scholar]
  23. Miceli M. V., Kan L. S., Newsome D. A. Phosphorus-31 nuclear magnetic resonance spectroscopy of human retinoblastoma cells: correlation with metabolic indices. Biochim Biophys Acta. 1988 Jul 29;970(3):262–269. doi: 10.1016/0167-4889(88)90125-5. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Okunieff P. G., Koutcher J. A., Gerweck L., McFarland E., Hitzig B., Urano M., Brady T., Neuringer L., Suit H. D. Tumor size dependent changes in a murine fibrosarcoma: use of in vivo 31P NMR for non-invasive evaluation of tumor metabolic status. Int J Radiat Oncol Biol Phys. 1986 May;12(5):793–799. doi: 10.1016/0360-3016(86)90038-6. [DOI] [PubMed] [Google Scholar]
  26. Okunieff P., Kallinowski F., Vaupel P., Neuringer L. J. Effects of hydralazine-induced vasodilation on the energy metabolism of murine tumors studied by in vivo 31P-nuclear magnetic resonance spectroscopy. J Natl Cancer Inst. 1988 Jul 20;80(10):745–750. doi: 10.1093/jnci/80.10.745. [DOI] [PubMed] [Google Scholar]
  27. Okunieff P., McFarland E., Rummeny E., Willett C., Hitzig B., Neuringer L., Suit H. Effects of oxygen on the metabolism of murine tumors using in vivo phosphorus-31 NMR. Am J Clin Oncol. 1987 Dec;10(6):475–482. doi: 10.1097/00000421-198712000-00003. [DOI] [PubMed] [Google Scholar]
  28. Okunieff P., Vaupel P., Sedlacek R., Neuringer L. J. Evaluation of tumor energy metabolism and microvascular blood flow after glucose or mannitol administration using 31P nuclear magnetic resonance spectroscopy and laser Doppler flowmetry. Int J Radiat Oncol Biol Phys. 1989 Jun;16(6):1493–1500. doi: 10.1016/0360-3016(89)90954-1. [DOI] [PubMed] [Google Scholar]
  29. Radda G. K., Rajagopalan B., Taylor D. J. Biochemistry in vivo: an appraisal of clinical magnetic resonance spectroscopy. Magn Reson Q. 1989 Apr;5(2):122–151. [PubMed] [Google Scholar]
  30. Rofstad E. K., DeMuth P., Fenton B. M., Sutherland R. M. 31P nuclear magnetic resonance spectroscopy studies of tumor energy metabolism and its relationship to intracapillary oxyhemoglobin saturation status and tumor hypoxia. Cancer Res. 1988 Oct 1;48(19):5440–5446. [PubMed] [Google Scholar]
  31. Rofstad E. K., DeMuth P., Sutherland R. M. 31P NMR spectroscopy measurements of human ovarian carcinoma xenografts: relationship to tumour volume, growth rate, necrotic fraction and differentiation status. Radiother Oncol. 1988 Aug;12(4):315–326. doi: 10.1016/0167-8140(88)90021-7. [DOI] [PubMed] [Google Scholar]
  32. Rofstad E. K., Fenton B. M., Sutherland R. M. Intracapillary HbO2 saturations in murine tumours and human tumour xenografts measured by cryospectrophotometry: relationship to tumour volume, tumour pH and fraction of radiobiologically hypoxic cells. Br J Cancer. 1988 May;57(5):494–502. doi: 10.1038/bjc.1988.113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rofstad E. K. NMR spectroscopy in prediction and monitoring of radiation response of tumours in vivo. Int J Radiat Biol. 1990 Jan;57(1):1–5. doi: 10.1080/09553009014550281. [DOI] [PubMed] [Google Scholar]
  34. Rofstad E. K., Wahl A., Stokke T., Nesland J. M. Establishment and characterization of six human melanoma xenograft lines. APMIS. 1990 Oct;98(10):945–953. doi: 10.1111/j.1699-0463.1990.tb05019.x. [DOI] [PubMed] [Google Scholar]
  35. Sijens P. E., Bovee W. M., Koole P., Schipper J. Phosphorus NMR study of the response of a murine tumour to hyperthermia as a function of treatment time and temperature. Int J Hyperthermia. 1989 May-Jun;5(3):351–357. doi: 10.3109/02656738909140461. [DOI] [PubMed] [Google Scholar]
  36. Sijens P. E., Bovée W. M., Seijkens D., Los G., Rutgers D. H. In vivo 31P-nuclear magnetic resonance study of the response of a murine mammary tumor to different doses of gamma-radiation. Cancer Res. 1986 Mar;46(3):1427–1432. [PubMed] [Google Scholar]
  37. Smith T. A., Eccles S., Ormerod M. G., Tombs A. J., Titley J. C., Leach M. O. The phosphocholine and glycerophosphocholine content of an oestrogen-sensitive rat mammary tumour correlates strongly with growth rate. Br J Cancer. 1991 Nov;64(5):821–826. doi: 10.1038/bjc.1991.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sostman H. D., Rockwell S., Sylvia A. L., Madwed D., Cofer G., Charles H. C., Negro-Vilar R., Moore D. Evaluation of BA1112 rhabdomyosarcoma oxygenation with microelectrodes, optical spectrophotometry, radiosensitivity, and magnetic resonance spectroscopy. Magn Reson Med. 1991 Aug;20(2):253–267. doi: 10.1002/mrm.1910200208. [DOI] [PubMed] [Google Scholar]
  39. Steen R. G. Characterization of tumor hypoxia by 31P MR spectroscopy. AJR Am J Roentgenol. 1991 Aug;157(2):243–248. doi: 10.2214/ajr.157.2.1853799. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Sutherland R. M., Rasey J. S., Hill R. P. Tumor biology. Am J Clin Oncol. 1988 Jun;11(3):253–274. doi: 10.1097/00000421-198806000-00004. [DOI] [PubMed] [Google Scholar]
  42. Tannock I. F., Rotin D. Acid pH in tumors and its potential for therapeutic exploitation. Cancer Res. 1989 Aug 15;49(16):4373–4384. [PubMed] [Google Scholar]
  43. Tozer G. M., Bhujwalla Z. M., Griffiths J. R., Maxwell R. J. Phosphorus-31 magnetic resonance spectroscopy and blood perfusion of the RIF-1 tumor following X-irradiation. Int J Radiat Oncol Biol Phys. 1989 Jan;16(1):155–164. doi: 10.1016/0360-3016(89)90023-0. [DOI] [PubMed] [Google Scholar]
  44. Tozer G. M., Griffiths J. R. The contribution made by cell death and oxygenation to 31P MRS observations of tumour energy metabolism. NMR Biomed. 1992 Sep-Oct;5(5):279–289. doi: 10.1002/nbm.1940050515. [DOI] [PubMed] [Google Scholar]
  45. Tozer G. M., Maxwell R. J., Griffiths J. R., Pham P. Modification of the 31P magnetic resonance spectra of a rat tumour using vasodilators and its relationship to hypotension. Br J Cancer. 1990 Oct;62(4):553–560. doi: 10.1038/bjc.1990.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Van der Grond J., Dijkstra G., Roelofsen B., Mali W. P. 31P-NMR determination of phosphomonoesters in relation to phospholipid biosynthesis in testis of the rat at different ages. Biochim Biophys Acta. 1991 May 24;1074(1):189–194. doi: 10.1016/0304-4165(91)90060-t. [DOI] [PubMed] [Google Scholar]
  47. Vaupel P., Kallinowski F., Okunieff P. Blood flow, oxygen and nutrient supply, and metabolic microenvironment of human tumors: a review. Cancer Res. 1989 Dec 1;49(23):6449–6465. [PubMed] [Google Scholar]
  48. Vaupel P., Okunieff P., Kallinowski F., Neuringer L. J. Correlations between 31P-NMR spectroscopy and tissue O2 tension measurements in a murine fibrosarcoma. Radiat Res. 1989 Dec;120(3):477–493. [PubMed] [Google Scholar]
  49. Vaupel P., Okunieff P., Neuringer L. J. In vivo 31P-NMR spectroscopy of murine tumours before and after localized hyperthermia. Int J Hyperthermia. 1990 Jan-Feb;6(1):15–31. doi: 10.3109/02656739009140801. [DOI] [PubMed] [Google Scholar]
  50. Wendland M. F., Iyer S. B., Fu K. K., Lam K. N., James T. L. Correlations between in vivo 31P MRS measurements, tumor size, cell survival, and hypoxic fraction in the murine EMT6 tumor. Magn Reson Med. 1992 Jun;25(2):217–232. doi: 10.1002/mrm.1910250202. [DOI] [PubMed] [Google Scholar]

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