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目的:探讨活化淋巴细胞中Survivin蛋白诱导表达的信号转导通路、分子级联反应和生物效应,揭示Survivin蛋白表达调控的分子机制。方法:用PHA和rhIL-2刺激培养人外周血单个核细胞,附加JAK抑制剂—AG490的处理,以Western blot检测Survivin和相关蛋白的表达;以流式细胞术(FCM)分析细胞周期和细胞分裂增殖。结果:刺激培养的细胞按照时序先后出现的分子和细胞学反应为:Stat3和Stat5磷酸化→CyclinD3、CyclinE蛋白水平上调→细胞进入S期及Survivin蛋白起始表达→细胞有丝分裂及Survivin蛋白表达水平增加。AG490对于上述反应均呈现明显的抑制作用,但对周期抑制蛋白P21和抗凋亡蛋白Bcl-2的表达水平没有影响。结论:Survivin蛋白的表达依赖JAK-Stat信号转导通路的早期活化,通过上调CyclinD3和CyclinE周期蛋白水平,启动细胞周期的运行,诱导Survivin蛋白的周期时相依赖性表达,参与细胞有丝分裂与增殖。
OBJECTIVE: To investigate the signal transduction pathway, molecular cascade and biological effects induced by survivin protein in activated lymphocytes and to reveal the molecular mechanism of Survivin protein expression. Methods: Human peripheral blood mononuclear cells (PBMCs) were stimulated with PHA and rhIL-2, and treated with JAK inhibitor-AG490. Survivin and related proteins were detected by Western blot. Cell cycle and cell cycle were analyzed by flow cytometry (FCM) Splitting proliferation. Results: The molecular and cytological responses of cultured cells stimulated by time sequence were as follows: Phosphorylation of Stat3 and Stat5 → Upregulation of CyclinD3 and CyclinE proteins → S phase entry into cells and initiation of Survivin protein → Increase in mitosis and Survivin protein expression . AG490 showed a significant inhibition on the above reaction, but had no effect on the expression level of P21 and Bcl-2. Conclusion: The expression of Survivin protein depends on the early activation of JAK-Stat signal transduction pathway. CyclinD3 and CyclinE cyclin levels are upregulated to activate the cell cycle and induce the cycle-dependent expression of Survivin protein, which is involved in cell mitosis and proliferation.