MC3T3-E1细胞流体剪切力敏感信号通路的基因芯片研究

目的 探讨适宜流体剪切力力值及作用时间下成骨细胞差异表达基因及相关信号通路。方法 通过平行板流室加力装置对盖玻片培养的MC3T3-E1细胞施加流体剪切力，提取总RNA，进行包括44 170个基因的全功能组表达谱基因芯片检测，对差异表达基因进行Pathway和GO分析，并采用实时荧光定量逆转录聚合酶链反应（RTPCR）RTPCR）对芯片结果进行验证。结果 芯片结果显示，差异基因共有884个，其中表达增强基因444个，表达降低基因440个。Pathway分析涉及的信号通路，主要包括Notch信号通路、RIG-Ⅰ样受体信号通路等。GO分析主要涉及前列腺素的生物合成、一氧化氮介导的信号传导、钙介导的信号及细胞免疫反应等不同的功能分类。结论 流体剪切力对MC3T3-E1细胞的保护作用可能与激活促进细胞生存及抑制细胞凋亡相关信号通路及生物学过程相关。

Genome-wide research on fluid shear stress-sensitive signaling pathways in MC3T3-E1 cells

Objective To explore the differentially expressed genes and related signaling pathways in MC3T3-E1 osteoblasts in response to suitable fluid shear stress values and action time with cDNA microarrays. Methods MC3T3-E1 cells cultured on a cover slip were subjected to fluid shear stress using a parallel plate flow chamber. The harvested RNA was used for microarray hybridization comprising approximately 44 170 genes, as well as for the subsequent real-time quantitative polymerase chain reaction validation of expression levels for selected genes. Microarray results were analyzed by using both GO and Pathway analysis. Results Microarray analysis indicated that 884 differentially expressed genes were found. Among these genes, 444 were upregulated, whereas 440 were downregulated. The Notch signal and RIG-Ⅰ-like receptor signaling pathways were involved in the Pathway analysis. GO analysis mainly involved different functional classifications, such as prostaglandin biosynthesis, nitric oxide-mediated signal transduction, calcium mediated signal, and cellular immune response, among others. Conclusion The mechanism underlying the protective effect of fluid shear stress on MC3T3-E1 cells might be related to promoting cell survival-and inhibiting cell apoptosis-related signaling pathways and biological processes.