14.7T核磁共振波谱评价脑胶质瘤细胞放射敏感性及放疗前后代谢物改变

发布时间：2018-11-26 07:45

【摘要】：研究目的：通过应用Bruker公司提供的Avance14.7T离体型高分辨率核磁共振波谱仪,探索体外脑胶质瘤细胞株放射敏感性与细胞内不同代谢物之间的关系。同时观测不同剂量X-线照射后肿瘤细胞凋亡情况及代谢物的改变。研究方法：利用高氯酸提取法从四种细胞株C6和9L,U251和T98G中获得细胞内水溶性代谢物样品,其中包括：肌酸(Cr),乳酸(Lac),N-乙酰天门冬氨酸(NAA),胆碱复合物(Cho),琥珀酸(Suc)等。将四种细胞内水溶性代谢物样品重新融于重水,并获得不同的核磁共振波谱。利用细胞集落形成实验测定四种胶质瘤细胞株的放射敏感性并计算出相应的放射敏感性参数。此外,将C6细胞株分别用OGy,4Gy,8Gy,12Gy,16Gy剂量的X-线照射,培养48小时后,从同一培养瓶内取出部分细胞检测其凋亡率,剩余的细胞用高氯酸提取法提取细胞内水溶性代谢物并获取核磁共振波谱。所有采集的核磁共振波谱数据最后均使用MestRe-c5.3软件进一步分析。研究结果：利用离体型高分辨率核磁共振波普仪可以测得质量优良的脑胶质瘤细胞水溶性代谢物波谱图像。体外脑胶质瘤细胞株放射敏感性与细胞内代谢物比值Cho/Cr, Cho/NAA呈正比关系,与Suc/Cr比值呈反比关系。C6细胞株经X-线照射后,细胞凋亡率和代谢物比值变化分别有特殊的下降期和平台期。结论：离体型高分辨率核磁共振波普能在细胞水平检测细胞内代谢物的微量改变,同时能够评价肿瘤细胞的放射敏感性。1H-MRS有望指导临床肿瘤放疗生物靶区的划定,进一步提高临床肿瘤放疗效果。
[Abstract]:Objective: to explore the relationship between radiosensitivity and intracellular metabolites of glioma cell lines in vitro by using Avance14.7T high resolution nuclear magnetic resonance spectrometer (NMR) provided by Bruker Company. At the same time, the apoptosis of tumor cells and the changes of metabolites were observed after different doses of X-ray irradiation. Methods: samples of intracellular water-soluble metabolites were obtained from four cell lines C6 and 9LU251 and T98G by perchloric acid extraction method, including: creatine (Cr), lactate (Lac), N-acetyl aspartate (NAA), Choline complex (Cho), succinate (Suc) et al. Four intracellular water-soluble metabolites were remelted into heavy water and different NMR spectra were obtained. The radiosensitivity of four glioma cell lines was measured by colony forming assay and the corresponding radiosensitivity parameters were calculated. In addition, C6 cells were irradiated with OGy,4Gy,8Gy,12Gy,16Gy dose of X- line for 48 hours, and some of the cells were extracted from the same culture flask to detect the apoptosis rate. The remaining cells were extracted with perchloric acid to extract the intracellular water-soluble metabolites and nuclear magnetic resonance (NMR) spectra were obtained. All the NMR data collected were further analyzed by MestRe-c5.3 software. Results: the high resolution nuclear magnetic resonance (NMR) spectrometer with high resolution in vitro was used to measure the high quality water-soluble metabolites of glioma cells. The radiosensitivity of glioma cell line in vitro was directly proportional to the ratio of intracellular metabolites (Cho/Cr, Cho/NAA) and inversely correlated with the ratio of Suc/Cr. The changes of apoptosis rate and metabolite ratio had special decline period and plateau stage, respectively. Conclusion: in vitro high resolution nuclear magnetic resonance imaging (MRI) can detect the microchanges of intracellular metabolites and evaluate the radiosensitivity of tumor cells at the cellular level. 1H-MRS is expected to guide the delineation of biological targets for clinical tumor radiotherapy. Further improve the clinical tumor radiotherapy effect.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R445.2;R739.41

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