摘要
目的用基因芯片技术分析不同分化程度的胶质瘤细胞凋亡相关的差异表达基因,为研究凋亡机制与胶质瘤细胞恶性程度关系奠定基础。方法用Trizol试剂盒抽提总RNA,用SuperscriptⅡ逆转录酶进行RT-PCR,荧光染料Cy3和Cy5标记cDNA产物;然后进行芯片杂交,检测人胶质瘤细胞系CHG-5(WHO分级为Ⅱ级)和SHG-44(WHO分级为Ⅳ级)瘤细胞基因表达的差异,特别是与细胞凋亡相关基因表达的差异。结果与CHG-5相比,SHG-44细胞中检测到有103个基因表达明显上调和89个基因表达明显下调,这些差异表达基因的种类很多,其中凋亡类相关基因共6个,上调基因有4个,下调基因2个。结论本结果提示,不同级别的胶质瘤可能有差异表达基因,特别是与细胞凋亡相关的差异表达基因。
Objective To identify the differential expression of the genes related to cell apoptosis in glioma cell lines, SHG-44 and CHG-5 by cDNA microarray in order to provide basic data for further research on the relationship between mechanisms of cell apoptosis and malignant degree of glio,nas. Methods Total RNAs of the cell lines, SHG-44 and CHG-5 was extracted by the Trizol reagents （Gibco-BRL）. RT-PCR was performed by reverse trascriptase, Superscript Ⅱ （Gibco-BRL）. The cDNA products were labeled with Cy3 and Cy5 fluorescence dye （Amersham Phannacia） for microarray hybridizations. Differential expression of genes of glioma cell lines, SHG-44 and CHG-5 were detected by cDNA microarray. Results Compared with the CHG-5 cell line, 103 genes were up-regulated （ratio〉2.0） and 89 genes were down-regulated （ratio〈0.5）, and of 6 genes related to apoptosis,3 were up-regulated and 3 down-regulated in the SHG-44 cell line. Furthmore, the differential expression of genes was confirmed by Northern blot hybridizations. Conclusion It is suggested that the differential expression of the genes, especially genes related to cell apoptosis may be involved in the progression of gliomas.
出处
《中国临床神经外科杂志》
2008年第10期611-613,共3页
Chinese Journal of Clinical Neurosurgery
基金
四川省卫生厅科研项目(050209)
关键词
胶质瘤
基因
凋亡
基因芯片
Glioma
Gene
Differential expression
Apoptosis
cDNA microarray
