Abstract
IntroductionGlioma is the most common type of the primary CNS tumor. Radiotherapy is an important treatment measure after surgery. However, its highly invasive character is the main reason of postoperative recurrence. The aim of the study was to probe the correlation between the invasion ability and the metabolite characteristics of glioma cells at the cellular level after irradiation by using 14.7T high-resolution nuclear proton magnetic resonance spectroscopy (1H-MRS). MethodsTo determine the matrix metalloproteinase-2 (MMP-2) activity and metabolite ratios of glioma cells after irradiation with different doses of X-rays, U87 and C6 glioma cells were exposed to X-ray irradiation of 0, 1, 5, 10, and 15Gy. After 20h, the perchloric acid (PCA) extraction method was used to evaluate water-soluble metabolites [choline (Cho), creatine (Cr), and N-acetylaspartate (NAA)], and 1H-MRS patterns and changes in metabolite ratios were observed in vitro by 14.7T high resolution 1H-MRS. Matrigel invasion assays and gelatin zymography were performed to test the invasion ability of U87 and C6 glioma cells. ResultsGood MR spectra were obtained from PCA method extracts of U87 and C6 glioma cells. Both radiation-induced MMP-2 activity and the Cho/Cr and Cho/NAA ratios increased after irradiation, and their increase occurred in a dose-dependent manner. The MMP-2 activity and the Cho/Cr and Cho/NAA ratios of glioma cells increased after irradiation up to 10Gy and decreased thereafter. In particular, the Cho/NAA ratio of U87 cells increased from 3.55±0.06 (0Gy) to 9.13±0.30 (10Gy) and then declined to 5.94±0.15 (15Gy). Furthermore, the invasion ability of glioma cells had a strong positive correlation with the Cho/Cr and Cho/NAA ratios. Both the Cho/Cr ratio and the Cho/NAA ratio of U87 glioma cells were highly positively correlated with the number of invading cells in the Matrigel invasion assay. The Pearson's correlation coefficient (r) values of U87 cells were 0.89 (Cho/Cr ratio versus invasion ability) and 0.91 (Cho/NAA ratio versus invasion ability) (P<0.01). C6 cells exhibited similar changes to those of U87 cells. ConclusionsIn vitro high-resolution 1H-MRS is useful for detecting glioma invasiveness at the cellular level.
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