少突胶质细胞电离辐射后早期基因表达谱的检测

目的 检测电离辐射后早期少突胶质细胞基因表达的变化特征.方法 将离体培养的少突胶质细胞经10 Gy照射后在1和4 h时间点,采用Affymetrix RAT 230 2.0基因表达谱芯片进行检测,并用实时荧光定量RT-PCR验证髓鞘碱性蛋白(MBP)和神经细胞间黏附分子1(NCAM-1)的测量结果.结果 少突胶质细胞经辐照后1和4 h内,其基因表达谱发生了明显的变化,这些基因有1079个已经命名并可以作功能分类.定量PCR表明MBP mRNA表达明显下调,而NCAM-1则上调,照射后4与1 h相比,表达量上调了3倍以上.结论 电离辐射后少突胶质细胞在几小时内就会发生一系列基因表达的变化,其中MBP和NCAM-1的改变可能在放射性脱髓鞘病变的发病机制中起重要作用.

Abstract：

ObjectiveTo characterize the gene expression in acute phase of irradiated oligodendrocytes (OL) in vitro.Methods The total RNA was extracted from irradiated OLs with 10 Gy by 6 MV X-rays at 1 and 4 h.The Affymetrix RAT 230 2.0 microarray were used to evaluate and screen the gene expression profile.The quantitative real-time RT-PCR was performed to validate the microarray results of selected myelin basic protein (MBP) and neural cell adhesion molecule 1 ( NCAM-1 ) genes.Results Compared with un-irradiated OLs,there were 1079 different expressed genes in irradiated cells.Those genes were classified in 79 categories based on the functional classification.Some familiar genes associated with OL cellular physiological process,apoptosis,cell cycle control,metabolism,cell communication and receptor binding were included.Compared with the microarray results,the coincidence rate of real-time RT-PCR was 91.7%.The down-regulation of MBP and up-regulation of NCAM 1 gene expression were confirmed.Conclusions Radiation-induced changes in gene expression in OLs took place in acute phase and influenced by time-course.The changes of MBP and NCAM1 gene expression may play a key role in the pathogenesis of radiation-induced demyelination.

Gene expression profile of oligodendrocytes in vitro in early stage after ionizing irradiation

Objective To characterize the gene expression in acute phase of irradiated oligodendrocytes (OL) in vitro. Methods The total RNA was extracted from irradiated OLs with 10 Gy by 6 MV X-rays at 1 and 4 h. The Affymetrix RAT 230 2.0 microarray were used to evaluate and screen the gene expression profile. The quantitative real-time RT-PCR was performed to validate the microarray results of selected myelin basic protein (MBP) and neural cell adhesion molecule 1 (NCAM-1) genes. Results Compared with un-irradiated OLs, there were 1079 different expressed genes in irradiated cells. Those genes were classified in 79 categories based on the functional classification. Some familiar genes associated with OL cellular physiological process, apoptosis, cell cycle control, metabolism, cell communication and receptor binding were included. Compared with the microarray results, the coincidence rate of real-time RT-PCR was 91.7%. The down-regulation of MBP and up-regulation of NCAM 1 gene expression were confirmed. Conclusions Radiation-induced changes in gene expression in OLs took place in acute phase and influenced by time-course. The changes of MBP and NCAM 1 gene expression may play a key role in the pathogenesis of radiation-induced demyelination.