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目的研究半胱氨酸蛋白酶(Caspase)激活方式,探讨Ⅲ型干扰素(IFNλ)信号通路诱导凋亡的机制。方法用IFNλ刺激人肺癌细胞A549,或用IL-10刺激表达复合受体FL-10R1/λR1的A549/FL-10R1/λR1细胞。用流式细胞分析检测Caspase3、Caspase8和Caspase9的激活程度。用Caspase抑制剂Z-VAD-FMK抑制Caspase,用流式细胞分析检测Caspase的激活情况,并通过末端转移酶标记术观察其对细胞凋亡的影响。结果 IFNλ信号通路可以上调Caspase3、Caspase8和Caspase9活化水平(5.5、3.5和4.5倍)并诱导凋亡。Z-VAD-FMK抑制Caspase3和Caspase8激活(60.0%和57.9%),但是提高了Caspase9的激活水平(35.1%),且不能阻止凋亡。结论 IFNλ信号可以激活不同的Caspase,通过激活死亡受体通路和线粒体通路来诱导凋亡,但是Caspase不是IFNλ诱导凋亡所必需的。
Objective To study the activation of Caspase and to explore the mechanism of apoptosis induced by type Ⅲ interferon (IFNλ) signal transduction pathway. Methods A549 / FL-10R1 / λR1 cells expressing human complex receptor FL-10R1 / λR1 were stimulated with IFNλ or stimulated with IL-10. Flow cytometry was used to detect the activation of Caspase3, Caspase8 and Caspase9. Caspase was inhibited by Caspase inhibitor Z-VAD-FMK. The activation of Caspase was detected by flow cytometry. The effect of Caspase on apoptosis was observed by telomerase activity assay. Results IFNλ signaling pathway can upregulate the activation of Caspase3, Caspase8 and Caspase9 (5.5, 3.5 and 4.5 fold) and induce apoptosis. Z-VAD-FMK inhibited Caspase3 and Caspase8 activation (60.0% and 57.9%), but increased Caspase9 activation (35.1%) and did not prevent apoptosis. Conclusion The IFNλ signal can activate different caspases and induce apoptosis through activation of death receptor pathway and mitochondrial pathway, but Caspase is not necessary for IFNλ-induced apoptosis.