小鼠促进骨髓抑制恢复相关因子IL-7基因的识别与克隆

The identification and cloning of IL-7 gene in myelosuppressed mice

目的寻找促进骨髓抑制恢复的相关细胞因子,进行骨髓抑制时基因表达差异分析,并对表达上调较大的细胞因子进行克隆验证。方法尾静脉注射5-Fu构建小鼠骨髓抑制模型,运用基因芯片方法分析正常小鼠与骨髓抑制模型第0,3,7,11,14天的基因表达差异,针对表达上调较大的鼠IL-7基因设计引物,以RT-PCR克隆其编码序列,并构建phCMV/IL-7真核表达载体。结果得到一组符合促进骨髓抑制恢复细胞因子特点的基因,其中细胞因子IL-7表达量升高30倍。结论IL-7符合促进骨髓抑制恢复细胞因子的特点,为研究骨髓抑制恢复的新方案奠定了基础。

Objective To find the cytokines that antagonized myelosuppression, and analyze gene expression in myelosuppressed mice and confirm remarkable upregnlation of gene expression. Methods The myelosuppressed mice model was made by injection of 5-Fu in tail veils. The gene differential expression in day 0, 3, 7, 11 and 14 respectively were analyzed with genechip. Specific primers of IL-7 which rose remarkably was designed to clone its full-length coding sequence （CDS） by RT-PCR, and then the eukaryotic vector phCMV/IL-7 was constructed. Results A group of cytokines that might play positive regulatory roles in suppression and regeneration of hematopoiesis were found. The expression level of IL- 7increased by 30 times compared with the normal control. Conclusions IL-7 was a potential upregulating cytokine, and might be clinically used for myelosuppression.