Magnetic resonance detects changes in phosphocholine associated with Ras activation and inhibition in NIH 3T3 cells.
Ras is frequently mutated in cancer, and novel therapies are being developed to target Ras signalling. To identify non-invasive surrogate markers of Ras activation and inhibition, we used(31)P magnetic resonance spectroscopy (MRS) and investigated NIH 3T3 cells compared to a mutant ras transfected counterpart. The MR spectra indicated that phosphocholine (PC) levels increased significantly from 3 +/- 2 fmol cell(-1)in NIH 3T3 cells to 13 +/- 4 fmol cell(-1)in the transfected cells. The PC/NTP ratio increased significantly from 0.3 +/- 0.1 to 0.7 +/- 0.3. This could not be explained by either a faster proliferation rate or by alterations in cell cycle distribution. Both cell lines were treated with simvastatin, 17-AAG and R115777, agents which inhibit Ras signalling. Cell proliferation was inhibited in both cell lines. The spectrum of NIH 3T3 cells was not affected by treatment. In contrast, in the ras transfected cells growth inhibition was associated with an average 35 +/- 5% drop in PC levels and a comparable drop in PC/NTP. Thus the MRS visible increase in phosphocholine is associated with Ras activation, and response to treatment is associated with partial reversal of phosphocholine increase in ras transfected cells. MRS might therefore be a useful tool in detecting Ras activation and its inhibition following targeted therapies.
Cancer Research Campaign (CRC) Clinical Magnetic Resonance Research Group, Institute of Cancer Research, Royal Marsden Hospital, Downs Road,Sutton, Surrey, SM2 5PT, UK.
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Ronen
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Jackson
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Beloueche
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Leach
D016475:3T3 Cells; D000818:Animals; D016227:Benzoquinones; D014408:Biomarkers, Tumor / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D004791:Enzyme Inhibitors / Q000494:pharmacology; D011905:Genes, ras; D047029:Lactams, Macrocyclic; D020935:MAP Kinase Signaling System / Q000187:drug effects; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D009154:Mutation; D009711:Nucleotides / Q000096:biosynthesis; D010767:Phosphorylcholine / Q000378:metabolism*; D016283:Proto-Oncogene Proteins p21(ras) / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015363:Quinolones / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives / Q000494:pharmacology; D019821:Simvastatin / Q000494:pharmacology; D014162:Transfection
D016227:Benzoquinones; D014408:Biomarkers, Tumor; D004791:Enzyme Inhibitors; D047029:Lactams, Macrocyclic; D009711:Nucleotides; D015363:Quinolones; D010767:Phosphorylcholine; D017828:Rifabutin; C112765:tanespimycin; D019821:Simvastatin; D016283:Proto-Oncogene Proteins p21(ras); C402769:tipifarnib