【摘要】：研究目的：通過應用Bruker公司提供的Avance14.7T離體型高分辨率核磁共振波譜儀,探索體外腦膠質(zhì)瘤細胞株放射敏感性與細胞內(nèi)不同代謝物之間的關(guān)系。同時觀測不同劑量X-線照射后腫瘤細胞凋亡情況及代謝物的改變。 研究方法：利用高氯酸提取法從四種細胞株C6和9L,U251和T98G中獲得細胞內(nèi)水溶性代謝物樣品,其中包括：肌酸(Cr),乳酸(Lac),N-乙酰天門冬氨酸(NAA),膽堿復合物(Cho),琥珀酸(Suc)等。將四種細胞內(nèi)水溶性代謝物樣品重新融于重水,并獲得不同的核磁共振波譜。利用細胞集落形成實驗測定四種膠質(zhì)瘤細胞株的放射敏感性并計算出相應的放射敏感性參數(shù)。此外,將C6細胞株分別用OGy,4Gy,8Gy,12Gy,16Gy劑量的X-線照射,培養(yǎng)48小時后,從同一培養(yǎng)瓶內(nèi)取出部分細胞檢測其凋亡率,剩余的細胞用高氯酸提取法提取細胞內(nèi)水溶性代謝物并獲取核磁共振波譜。所有采集的核磁共振波譜數(shù)據(jù)最后均使用MestRe-c5.3軟件進一步分析。 研究結(jié)果：利用離體型高分辨率核磁共振波普儀可以測得質(zhì)量優(yōu)良的腦膠質(zhì)瘤細胞水溶性代謝物波譜圖像。體外腦膠質(zhì)瘤細胞株放射敏感性與細胞內(nèi)代謝物比值Cho/Cr, Cho/NAA呈正比關(guān)系,與Suc/Cr比值呈反比關(guān)系。C6細胞株經(jīng)X-線照射后,細胞凋亡率和代謝物比值變化分別有特殊的下降期和平臺期。 結(jié)論：離體型高分辨率核磁共振波普能在細胞水平檢測細胞內(nèi)代謝物的微量改變,同時能夠評價腫瘤細胞的放射敏感性。1H-MRS有望指導臨床腫瘤放療生物靶區(qū)的劃定,進一步提高臨床腫瘤放療效果。
[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.
【學位授予單位】：山東大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R445.2;R739.41

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