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目的 克隆胶质瘤细胞诱导分化相关新基因。方法 应用RNA随机引物差异显示、SSCP纯化、PCR产物快速克隆、反Northern杂交及生物信息学分析等方法 ,观察人脑胶质瘤细胞株SHG 44 9分化过程中的基因表达变化。结果 克隆了诱导分化前后表达差异显著的 1 5个基因 ,其中诱导后下调基因 4个 ,上调基因 1 1个。同源性分析表明 ,5个是已知基因 ,1 0个是未知基因。诱导后上调的DIG 1基因与c myc内含子结合蛋白 1 (MIBP1 )基因高度同源 ,此基因具有转录因子活性 ,表达的蛋白可抑制c myc基因的表达。结论 克隆了 1 5个人脑胶质瘤细胞诱导分化相关基因 ,其中控制c myc基因表达的MIBP1基因诱导分化后上调提示 ,它可能是调控胶质瘤细胞分化基因 ,值得进一步研究。
Objective To clone new genes related to differentiation of glioma cells. Methods The gene expression changes in SHG 44 9 human glioma cell line SHG 44 9 were observed by differential display of random primers, SSCP purification, rapid cloning of PCR products, reverse Northern hybridization and bioinformatics analysis. Results Fifteen differentially expressed genes were differentially expressed before and after differentiation, including 4 down-regulated genes and 11 up-regulated genes. Homology analysis showed that 5 were known genes and 10 were unknown genes. The up-regulated DIG1 gene is highly homologous to the c-myc intron-binding protein 1 (MIBP1) gene after induction. This gene has the transcription factor activity, and the expressed protein can inhibit the expression of c-myc gene. Conclusion Cloning of 15 human glioma cell differentiation-related genes, which control the c-myc gene expression of MIBP1 gene induced differentiation prompted, it may be regulation of glioma cell differentiation gene, worthy of further study.