应用基因表达谱芯片研究内皮抑素作用后人脐静脉内皮细胞株基因表达的变化  

Gene expression of ECV304 cells after treated with endostatin detected by microarray analysis

作　　者：杨获[1] 张美霞[1] 张军军[1] 严密[1] 

机构地区：[1]四川大学华西医院眼科,成都610041

出　　处：《中华眼底病杂志》2010年第4期353-356,共4页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金（30500547）

摘　　要：目的 观察人脐静脉内皮细胞株（ECV304细胞）在内皮抑素（ES）作用前后基因表达谱的变化，并探讨其作用的分子机制。方法ES处理培养的ECV304细胞36h，抽提ES作用前后细胞mRNA，用Cy3和Cy5不同的荧光标记脱氧尿嘧啶核苷三磷酸（dUTP）逆转录制备cDNA探针，与载有4096条基因的基因芯片进行杂交，通过对Cy3、Cy5荧光信号扫描后，分析经ES处理前后基因水平表达差异。结果ES处理后细胞增生减慢，抽提RNA电泳18S、28S条带清晰，吸光度[A，旧称光密度（OD）]值为2．0-2．2。芯片扫描显示5．96％的检测基因出现明显表达差异，其中下调基因225条，上调基因19条。结论ECV304细胞经ES作用后部分基因表达呈下调趋势，可能与ES抑制新生血管生成机制有关。Objective To observe the altered gene expression of ECV304 cell after treated with endostatin （ES） using cDNA microarray, and to investigate its molecular mechanism. Methods BiostarH- 40S gene expression DNA mieroarrays were used to profile changes in gene expression of ECV304 cells after treated with ES for 36 hours. To prepare the probes, mRNA from both control and treated cells were isolated and purified, and then reversely transcribed to cDNA with the incorporation of fluorescent-labeled dUTP： Cy3 and Cy5 respectively. The probes were hybridized with expressed cDNA mieroarray, the fluorescent signals of Cy3 and Cy5 were scanned and analyzed by a computer system. Results ES inhibited the proliferation of ECV304 cells. The result of electrophoresis indicated that the extracted total RNA had high quality. Approximately 5.96% of all human genes examined on 4096 gene microarrays showed altered expression, with 225 down-regulated genes and 19 up-regulated genes, Conclusion The expression of some genes of ECV304 cells decrease after treated with ES, which may be related to the mechanism of inhibition of neovaseularization.

关 键 词：基因表达谱/利用 芯片分析技术/方法 内皮抑素类/药理学 基因表达 

分 类 号：R774.1[医药卫生—眼科] R349.64[医药卫生—临床医学]