Analysis of antiapoptotic genes induced in vascular endothelial cells by monocyte adhesion.

血管内皮细胞的凋亡和单核细胞与内皮细胞的相互作用在动脉粥样硬化的发生发展中起着至关重要的作用。我们发现单核细胞与EC的黏附抑制了BC的凋亡。然而，调控BC细胞凋亡的机制仍有待阐明。在本研究中，我们使用人单核细胞系THP-1和人脐静脉内皮细胞(HUVEC)分析了单核细胞黏附诱导BC细胞凋亡的基因。用血清饥饿诱导HUVEC凋亡，用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法和caspase-3活性检测。细胞凋亡。THP-1黏附对HUVEC有明显抑制作用。采用基因芯片技术分析THP-1与HUVEC黏附诱导的基因表达。THP-1与HUVEC的黏附诱导HUVEC表达多种基因，包括骨桥蛋白、IL-1、选择素B、PPAR-γ和MCP-1，以及与凋亡相关的基因bcl2、cspace-8和BIK。在这些表达的基因中，骨桥蛋白最显著地被THP-1黏附上调(增加了558倍)。因为骨桥蛋白已被报道在相关的生存通路中发挥作用。在细胞黏附方面，我们的研究结果为单核细胞黏附抑制内皮细胞凋亡的遗传机制提供了新的视角。

Apoptosis of vascular endothelial cells (EC) and monocyte-EC interaction play a critical role in the development and progression of atherosclerosis. We have found that adhesion of monocyte to EC inhibits BC apoptosis. However, the mechanism by which BC apoptosis is regulated remains to be elucidated. In the present study, we analyzed antiapoptotic genes induced in BC by monocyte adhesion using THP-1, a human monocytic cell line and human umbilical vein endothelial cells (HUVEC). Apoptosis of HUVEC was induced by serum starvation arid examined by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) labeling and caspase-3 activity assay. Apoptosis of. HUVEC was significantly inhibited by THP-1 adhesion. mRNAs were isolated from HUVEC cultured with or without THP-1, and genes induced in HUVEC by THP-1 adhesion were analyzed using a DNA microarray (IntelliGene HS Human Expression CHIP, TAKARA BIO). Adhesion of THP-1 to HUVEC induced expression of many kinds of genes in HUVEC including osteopontin, IL-1, selectin B, PPARgamma, and MCP-1 as well as apoptosis-relaled genes such as bcl-2, caspace-8 and BIK. Among these genes expressed, osteopontin was most prominently upregulated by THP-1 adhesion (558-fold increase). Because osteopontin has been reported to play a role in survival pathways related. to cell adhesion, our data provide a novel insight into the genetic mechanism of inhibition of EC apoptosis induced by monocyte adhesion.