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核转录因子-κB(NF-κB)是维持急性淋巴细胞白血病(ALL)细胞生存的关键因子.近年来发现,糖原合成酶激酶-3β(GSK-3β)可以正性调控NF-κB的活性.本研究通过抑制GSK-3β活性初步探讨ALL细胞中GSK-3β在NF-κB诱导细胞凋亡中的作用机制.收集ALL患儿骨髓单个核细胞,采用免疫荧光细胞化学方法检测到ALL细胞核内GSK-3β有明显聚集.体外培养ALL细胞后经GSK-3β抑制剂氯化锂(LiCl)和SB216763处理,采用Western印迹和EMSA检测发现,ALL细胞核内GSK-3β表达下降,而NF-κBP65蛋白无明显变化,但是其活性明显降低.同时RT-PCR结果显示,NF-κB下游抗凋亡基因存活素(survivin)的表达随之下降,AnnexinV-PE/7-AAD双染流式细胞仪检测结果证实,ALL细胞凋亡明显增加(P<0.01).该结果表明,抑制GSK-3β活性可以下调NF-κB的转录活性,并通过下调抗凋亡基因存活素的表达而促进ALL细胞的凋亡.
Nuclear factor-kappa B (NF-κB) is a key factor in maintaining the survival of acute lymphoblastic leukemia (ALL) cells. In recent years, it has been found that glycogen synthase kinase-3β (GSK-3β) can positively regulate the activity of NF- In this study, the mechanism of GSK-3βin NF-κB-induced apoptosis in ALL cells was initially investigated by inhibiting the activity of GSK-3β. Bone marrow mononuclear cells (ALL) were collected from ALL patients and detected by immunofluorescence cytochemistry GSK-3βis significantly aggregated.After cultured with ALL cells in vitro, the GSK-3β inhibitor LiCl and SB216763 were treated by Western blot and EMSA, and the expression of GSK-3β in ALL cells was decreased, whereas the expression of NF- However, its activity was significantly decreased, while the expression of survivin, an inhibitor of downstream NF-κB gene, was down-regulated by RT-PCR. The expression of survivin mRNA in the downstream of NF-κB was detected by flow cytometry with Annexin V-PE / 7-AAD The results showed that the apoptosis of ALL cells was significantly increased (P <0.01) .This result shows that inhibiting the activity of GSK-3β can down-regulate the transcriptional activity of NF-κB and promote the apoptosis of ALL cells by down-regulating the expression of anti-apoptotic gene survivin Death.