利用基因芯片技术研究造血生长因子在脐静脉内皮细胞中的表达

为探讨利用基因芯片技术研究脐静脉内皮细胞中造血生长因子基因的表达情况，将体外培养的人脐静脉内皮细胞(ECV304)分为血管内皮生长因子(VEGF)处理组(添加50ng／ml的rhVEGF165)和正常对照组；培养24小时后收获细胞，提取总RNA，制备Cy3-dUTP标记的对照组cDNA探针和Cy5-dUTP标记的VEGF组cDNA探针，并与表达谱芯片杂交。杂交信号经荧光共聚焦显微镜扫描后，计算机分析基因表达情况。结果显示。两组细胞均表达多种造血生长因子及受体如Epo/R、GM—CSF／R、G—CSF／R、LIF、IL-3、TPO、Fit-3和SCF；同时也表达多种生长因子(VEGF、IGF2、PDGFA、PDGFB、TGFβ1)和生长因子受体(神经纤维黏蛋白-1，神经纤维黏蛋白-2，TGFβ-R1)；此外，共有24个基因存在差异表达。结论：利用基因芯片技术能够在短时间内迅速而准确地分析脐静脉内皮细胞众多造血生长因子及其受体的表达情况，为进一步研究内皮细胞的生物学特性提供了有效手段。

Study on the Expression of Hematopoietic Growth Factor Gene in Human Umbilical Vein Endothelial Cells Using Gene Chip

In order to detect the hematopoietic growth factor gene expressed in human umbilical vein endothelial cells using gene chip, human umbilical vein endothelial cells (ECV304) were cultured in vitro and divided into VEGF group and control group in same medium. 50 ng/ml hVEGF165 was added in the VEGF group. After culture for 24 hours all cells were collected for total RNA extraction. Then, cDNAs were marked with Cy3 and Cy5 for control group and VEGF group, respectively, and hybridized with gene chip. Hybridization signals were collected and analyzed following scanning by laser co-focal microscopy. The results showed that a large number of hematopoietic growth factor and receptor genes (Epo/R, GM-CSF/R, G-CSF/R, LIF, IL-3, TPO, Flt-3, SCF) were expressed in both groups, while many other growth factors (VEGF, IGF2, PDGFA, PDGFB, TGFbeta1) and receptors (neuropilin-1, neuropilin-2, TGFbeta-R1)were expressed. The differentially expressed genes amounted to 24. It is concluded that many hematopoietic growth factors and receptors expressed by hUVECs could be analyzed in a short period by using gene chip, which provides a powerful method for further studies on characteristics of vascular endothelial cells.