文档介绍：SBC基因芯片研究骨肉瘤发病转移相关基因及其发现
作者:曹磊,纪方,蔡晓冰,唐昊,竺伟,张瑞
【摘要】[目的] 运用基因芯片技术探索骨肉瘤发病及转移相关基因,研究基因表达水平对于骨肉瘤发生及转移的重要意义。[方法] 采用2个公认的骨肉瘤细胞株及1个成骨细胞株,提取总RNA,合成生物素标记的cRNA与SBC基因芯片杂交。随机挑选4个差异表达基因,分别设计合成引物,用嵌合荧光法(SYBR Green I)实时RT-PCR法定量测量术中收集的新鲜骨肉瘤标本及两株细胞中各基因的表达水平,应用ABI Prism 7 300分析其与成骨细胞株之间的表达差异。[结果]以表达差异≥≤,在SBC芯片包含的所有基因(约15 000个转录本)中,2个骨肉瘤细胞株与成骨细胞株相比较,共同上调189个基因;共同下调84个基因。随机挑选4个差异表达基因的实时RT-PCR结果与芯片结果完全相符。[结论]骨肉瘤是一种多基因病变,应用基因芯片技术有助于发现该肿瘤的基因变化规律。
【关键词】骨肉瘤; 基因微矩阵; 基因
Abstract: [Objective]To investigation the relavant genes to the pathogenesis and metastasis of a,and to discuss the significance of gene expression in the pathogenesis and metastasis of a. [Method]Two well-known a cell lines and an osteoblastic cell line were collected. After the total RNA was extracted,the finally synthesized biotinylated cRNAs were hybridized to SBC the design and synthethesis of the primers,four of the differentially expressed genes were then chosen at
random to further confirm the array results using the SYBR Green real-time RT-PCR method in freshly collected a specimens and the cell data analysis depended on the ABI Prism 7300 system.[Result]By an entrance limit of ≥ or ≤, 189 up-regulated and 84 dowm-regulated genes which were considerable differentially expressed in the two a cell pared with the osteoblastic cell line were finally of the genes were first reported to be related to the pathogenesis of a. The array results were further confirmed by the real-time RT-PCR.[Conclusion]Many genes are involved in the pathogenesis and metastasis of a. It is helpful to apply gene array to find the laws of gene expression and to study the gene-gene interactional relationships in this tumor.
Key words:a; gene microarray; gene
目前研究认为,骨肉瘤的发生与癌基因激活,抑癌基因失活及其它基因的表达失控有关,是多种相关基因多阶段多途径协同作用的结果。基因芯片又称DNA芯片或DNA微阵列,该技术由于采用了微电子学的并行处理和高密度集成的概念,具有高效、高信息量的突出优点。本实验以骨肉瘤细胞系(MG-63,U-2OS),应用SBC的人16K基因表达谱cDNA芯片(含已知基因12581个),通过一次重复,鉴定已知基因在骨肉瘤细