应用差异显示技术克隆胶质瘤细胞诱导分化相关基因

Clone the genes associated with induced differentiation of glioma with RAP-DD

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 the genes associated with induced differentiation of glioma.Methods RNA arbitrary primer differential display, SSCP, PCR product fast cloning, reverse Northern blot, sequencing and bioinformatics were used to study the differentiation of human glioma cell strain SHG 44 9 induced by Sodium Phenylbutyrate. Results 15 differential expressed genes in this process were screened out and cloned, in which 4 genes are with down regulated, function and the others with are are up regulated function. Bioinformatics analysis showed. 10 genes are unknown and 5 genes are known, including MIBP1, which is up regulated, whose protein can suppress c myc expression. Conclusions 15 genes associated with induced differentiation of glioma were cloned. In which, MIBP1 gene was up regulated after differentiation inducing, which control c myc expression, may play role in regulation of differentiation of glioma cells.