摘要
目的应用高密度Oligo基因芯片技术检测榄香烯对胶质瘤U251细胞凋亡相关基因的影响。方法实验分2组,实验组用含有终浓度为0.062mg/ml榄香烯作用于U251细胞24h,对照组为未行任何处理的U251细胞。抽提2组U251细胞的总RNA并逆转录合成Cy5、Cy3标记的cDNA,混合后杂交含22000个人类基因的人类高密度Oligo基因芯片,经过洗片和扫描,获得荧光信号图像并用计算机分析,随机抽取4种差异表达基因用PCR验证。结果按差异显著性标准,从22000条基因中筛选出差异表达基因179条,其中11个凋亡相关基因表达上调,15个凋亡相关基因表达下调。主要包括细胞周期蛋白、细胞凋亡、细胞骨架和运动蛋白等相关基因。结论榄香烯能够影响胶质瘤细胞相关凋亡基因的改变,而基因芯片技术有效分析其基因表达谱,有助于认识胶质瘤药物治疗的机制。
Objective To study the effect of elemene on apoptosis-associated genes in glioma U251 cell by the large-scale Oligo microarray. Methods The study was divided into two groups： control group, and treatment groups with elemene in human glioma U251 after 24h. The total RNAs were isolated from the two groups, and were reversely transcribed to cDNAs and labeled with the fluorescent Cy5-dNTP and Cy3-dNTP to prepare the hybridization probes. The mixed probes were hybridized to the large-scale Oligo microarray of 22 000 genes. After washing and scan, the fluorescent image was obtained and analyzed by computer. Results By applying the large-scale oligo microarray, 179 differentially expressed genes in glioma were screened out among the 22 000 target genes, comprising 11 upregulated genes and 15 downregulated apoptosis-associated genes respectively, including early response genes. Bcl-2 family genes, caspase family genes and cell cycle-associated genes. Conclusion Elemene has a effect of elemene on apoptosis-associated genes in glioma U251 cell, and Oligo microarray could analyede the screening and knowleded the mechanism of the glioma by drug.
出处
《福建医药杂志》
CAS
2007年第5期102-105,共4页
Fujian Medical Journal
基金
福建省科技厅科技计划项目(2001Z032)
