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目的构建胶质瘤细胞系U87的多细胞球体(MCS)模型,探讨不同剂量的X线对胶质瘤多细胞球体上清液基质金属蛋白酶2(MMP-2)活性和水溶性代谢物的影响。方法采用琼脂糖培养法建立U87多细胞球体模型;用0、1、5、10 Gy的X线分别照射多细胞球体,0 Gy组作为对照组,利用高氯酸提取细胞中水溶性代谢物[胆碱(Cho)、肌酸(Cr)、N-乙酰天门冬氨酸(NAA)],采用14.7T高场强磁共振仪获取水溶性代谢物波谱;采用明胶酶谱法测量U87多细胞球体上清中MMP-2活性。结果成功建立了U87多细胞球体模型;多细胞球体在1、5、10 Gy X线照射下的胆碱与肌酸比值(Cho/Cr)分别为2.74±0.02、4.32±0.17、4.92±0.07,与对照组(2.10±0.29)比较,细胞内水溶性代谢物比值Cho/Cr升高(P=0.033,P=0.000,P=0.000);多细胞球体在0~10 Gy X线照射范围内,随着照射剂量的增加,细胞M M P-2活性增加。与对照组比较,差异有统计学意义(P=0.000)。结论亚致死剂量的X线可导致U87胶质瘤多细胞球体MMP-2活性增高,Cho/Cr升高。
Objective To construct a multicellular spheroid (MCS) model of glioma cell line U87 and investigate the effects of different doses of X-ray on the activity of matrix metalloproteinase 2 (MMP-2) and water-soluble metabolites in glioma cell supernatant . Methods The U87 multi-spheroid model was established by agarose culture. The spheroids of 0, 1, 5, and 10 Gy were irradiated with multi-spheroids and the 0 Gy group was used as the control group. Perchloric acid was used to extract water-soluble metabolites [ Cho, creatine and NAA were measured. The spectra of water-soluble metabolites were obtained by 14.7T high-field-intensity magnetic resonance spectroscopy. U87 multicellular spheroids Supernatant MMP-2 activity. Results The U87 multi-spheroid model was established successfully. The ratio of choline to creatine (Cho / Cr) of multicellular spheroids at irradiation of 1, 5 and 10 Gy was 2.74 ± 0.02, 4.32 ± 0.17 and 4.92 ± 0.07, Compared with the control group (2.10 ± 0.29), the ratio of intracellular water soluble metabolites Cho / Cr increased (P = 0.033, P = 0.000, P = 0.000) As the radiation dose increased, the activity of MM P-2 increased. Compared with the control group, the difference was statistically significant (P = 0.000). Conclusion Sublethal dose of X-ray can lead to increased MMP-2 activity and increased Cho / Cr in U87 glioma multicellular spheroids.