左旋门冬酰胺酶对伯基特淋巴瘤细胞株增殖、细胞周期及凋亡的影响

Effects of asparaginase on proliferation,cell cycle and apoptosis of Burkitt lymphoma cell lines

目的探讨左旋门冬酰胺酶对伯基特淋巴瘤细胞株增殖、细胞周期和凋亡的影响及其作用机制。方法应用CCK-8法检测左旋门冬酰胺酶对伯基特淋巴瘤细胞株细胞增殖的影响,流式细胞术检测细胞凋亡率和细胞周期,实时定量PCR和Western blot检测分析细胞周期、凋亡、自噬和PI3K/Akt/mTOR信号通路中各种分子的表达变化。结果左旋门冬酰胺酶明显抑制多种伯基特淋巴瘤细胞株的增殖,并引起细胞周期G_(0)/G_(1)期阻滞,诱导细胞凋亡和自噬。进一步结果表明左旋门冬酰胺酶抑制c-Myc的表达,同时抑制p-PI3K、p-Akt-S473、p-mTOR、p-70S6K和p-4E-BP1的表达。PI3K抑制剂LY294002联合左旋门冬酰胺酶进一步诱导了细胞凋亡。此外,左旋门冬酰胺酶抑制STAT和ERK信号通路。结论左旋门冬酰胺酶抑制伯基特淋巴瘤细胞株细胞增殖和G_(0)/G_(1)期细胞阻滞,诱导自噬和凋亡并通过抑制PI3K/Akt/mTOR信号通路调控细胞凋亡。

Objective To investigate the effect of L-asparaginase on the proliferation,cell cycle,and apoptosis of Burkitt lymphoma cell lines and explore the molecular mechanism.MethodsThe effectof L-asparaginase on the cell proliferation of Burkitt lymphoma cell lines was detected using the CCK-8 method.The apoptosis rate and cell cycle were detected using flow cytometry.The expression of relatedmolecules in cell cycle,apoptosis,autophagy,and PI3K/Akt/mTOR signaling pathway was detected andanalyzed using qPCR and Western blot assay.Results L-asparaginase significantly inhibited theproliferation of Burkitt lymphoma cell lines and caused cell cycle arrest at G_(0)/G_(1),phage.L-asparaginaseinduced cell apoptosis and autophagy in Burkitt lymphoma cell lines.Further results showed thatL-asparaginase inhibited the expression of c-Myc and also inhibited the expression of p-PI3K,p-Akt-S473,p-mTOR,p-70S6K,and p-4E-BP1.Combining PI3K inhibitor LY294002 with L-asparaginase furtherinduced apoptosis.Additionally,L-Asp inhibited STAT and ERK signaling pathways.Conclusion L-asparaginase inhibited Burkit lymphoma cell proliferation,arrested cell cycle,activated autophagy,andinduced apoptosis by inhibiting the PI3K/Akt/mTOR signaling pathway.