Mowa Chishimba N, Li Tianbo, Jesmin Subrina, Folkesson Hans G, Usip Sharon E, Papka Raymond E, Hou Guichuan
Appalachian State University, Department of Biology, Boone, NC 28608, USA.
Reprod Biol Endocrinol. 2008 Dec 16;6:64. doi: 10.1186/1477-7827-6-64.
VEGF-regulated genes in the cervices of pregnant and non-pregnant rodents (rats and mice) were delineated by DNA microarray and Real Time PCR, after locally altering levels of or action of VEGF using VEGF agents, namely siRNA, VEGF receptor antagonist and mouse VEGF recombinant protein.
Tissues were analyzed by genome-wide DNA microarray analysis, Real-time and gel-based PCR, and SEM, to decipher VEGF function during cervical remodeling. Data were analyzed by EASE score (microarray) and ANOVA (Real Time PCR) followed by Scheffe's F-test for multiple comparisons.
Of the 30,000 genes analyzed, about 4,200 genes were altered in expression by VEGF, i.e., expression of about 2,400 and 1,700 genes were down- and up-regulated, respectively. Based on EASE score, i.e., grouping of genes according to their biological process, cell component and molecular functions, a number of vascular- and non-vascular-related processes were found to be regulated by VEGF in the cervix, including immune response (including inflammatory), cell proliferation, protein kinase activity, and cell adhesion molecule activity. Of interest, mRNA levels of a select group of genes, known to or with potential to influence cervical remodeling were altered. For example, real time PCR analysis showed that levels of VCAM-1, a key molecule in leukocyte recruitment, endothelial adhesion, and subsequent trans-endothelial migration, were elevated about 10 folds by VEGF. Further, VEGF agents also altered mRNA levels of decorin, which is involved in cervical collagen fibrillogenesis, and expression of eNO, PLC and PKC mRNA, critical downstream mediators of VEGF. Of note, we show that VEGF may regulate cervical epithelial proliferation, as revealed by SEM.
These data are important in that they shed new insights in VEGF's possible roles and mechanisms in cervical events near-term, including cervical remodeling.
在使用VEGF试剂（即小干扰RNA、VEGF受体拮抗剂和小鼠VEGF重组蛋白）局部改变VEGF水平或作用后，通过DNA微阵列和实时定量PCR对妊娠和未妊娠啮齿动物（大鼠和小鼠）子宫颈中的VEGF调控基因进行了描绘。
通过全基因组DNA微阵列分析、实时定量和基于凝胶的PCR以及扫描电子显微镜对组织进行分析，以解读子宫颈重塑过程中VEGF的功能。数据通过EASE评分（微阵列）和方差分析（实时定量PCR）进行分析，随后进行Scheffe氏F检验以进行多重比较。
在分析的30000个基因中，约4200个基因的表达因VEGF而改变，即分别约有2400个和1700个基因的表达下调和上调。基于EASE评分，即根据基因的生物学过程、细胞成分和分子功能对基因进行分组，发现子宫颈中许多与血管和非血管相关的过程受VEGF调控，包括免疫反应（包括炎症）、细胞增殖、蛋白激酶活性和细胞粘附分子活性。有趣的是，一组已知或有可能影响子宫颈重塑的特定基因的mRNA水平发生了改变。例如，实时定量PCR分析表明，VEGF使白细胞募集、内皮细胞粘附及随后的跨内皮迁移中的关键分子VCAM-1的水平升高了约10倍。此外，VEGF试剂还改变了参与子宫颈胶原纤维形成的核心蛋白聚糖的mRNA水平，以及VEGF关键下游介质eNO、PLC和PKC mRNA的表达。值得注意的是，扫描电子显微镜显示VEGF可能调节子宫颈上皮细胞增殖。
这些数据很重要，因为它们为VEGF在近期子宫颈事件（包括子宫颈重塑）中可能的作用和机制提供了新的见解。
