小鼠p27Kip1基因的克隆及与外周B细胞凋亡和生存的关系

目的：克隆小鼠的p27^kip1基因，并以此为探针分析p27^kip1在外周血B细胞中的表达，以及其在抗原受体和CD40介导的抑制和促进增殖信号中的作用。方法：从培养的B细胞中提取总RNA，用RT-PCR扩增并克隆p27^kip1基因，用流式细胞仪检测B细胞凋亡，用Northern blot观察p27^kip1mRNA的表达水平。结果：成功地克隆了小鼠的p27^kip1基因，发现抗原受体交联在引起外周血B细胞凋亡之前，出现p27^kip1表达上调，若同时激活CD40则使p27^kip1表达降低，B细胞也免于凋亡用针对p27^kip1的反义寡核苷酸处理，可阻断抗原受体交联引起的外周血B细胞凋亡。结论：p27^kip1可能在抗原受体信号介导的B细胞凋亡和CD40信号介导的B细胞生存中发挥一定的作用。

Cloning of mouse p27Kipl and investigation on its role in apoptosis and survival of peripheral blood B lymphocytes

AIM: To clone the mouse p27Kipl gene and analyze its expression and possible function in the signaling through antigen receptor and CD40 in peripheral blood B cells. METHODS; Mouse p27Kipl was cloned from cultured B cells by RT-PCR. Apoptosis of the B cells was detected by flow cytometry. Expression of p27Kipl gene was detected by Northern blot. RESULTS: Mouse p27Kipl gene was successfully cloned. In consistence with B cell apoptosis or survival, cross-linking of antigen receptor also induced up-regulation of p27Kipl in mouse B cells, which was blocked by CD40 signaling. Antigen receptor-mediated B cell apoptosis was blocked by anti-sense oligonucleotides directed against p27Kipl. CONCLUSION; These data suggest that p27Kipl may be involved in apoptosis and survival of B cells mediated by antigen receptor and CD40 signals, respectively.