LY294002对Jeko-1细胞的增殖抑制及作用机制研究

Antiproliferative Effects of LY294002 on MCL Jeko-1 Cell Line and Its Mechanism

本研究旨在探讨磷脂酰肌醇3-激酶/丝氨酸苏氨酸激酶(PI3K/Akt)特异性抑制剂LY294002对人套细胞淋巴瘤Jeko-1细胞生长、细胞凋亡的影响及其作用机制。四甲基偶氮唑盐(MTT)方法检测细胞增殖率;流式细胞术检测细胞凋亡水平;Western blot检测凋亡相关蛋白Cyclin D1、Bcl-2、procaspase-3及PI3K/Akt信号通路相关蛋白p-Akt、p-mTOR、p-P70S6K的变化。结果表明,LY294002能抑制Jeko-1细胞的增殖。Jeko-1细胞经LY294002 0、5、10和20μmol/L作用24 h后,凋亡率分别为(3.25±1.27)%、(11.34±2.35)%、(22.81±2.74)%和(43.61±3.48)%,差异有统计学意义(P<0.01);Western blot检测发现,凋亡相关蛋白Cyclin D1、Bcl-2、procaspase-3表达下降,PI3K/Akt信号通路相关蛋白p-Akt、p-mTOR、p-P70S6K磷酸化水平降低。结论:LY294002可明显抑制Jeko-1细胞增殖,其机制可能通过下调PI3K/Akt信号通路中的p-Akt、p-mTOR、p-P70S6K磷酸化水平,抑制PI3K/Akt信号通路,促进细胞凋亡。

This study was aimed to investigate the effects of LY294002, a specific inhibitor of phosphatidylinositol 3- kinase, on growth and apoptosis of MCL Jeko-1 cell line and its mechanism. The proliferation inhibitory rate of Jeko-1 cells treated by different doses of LY294002 was assayed by MTT method; the level of apoptosis of Jeko-1 cells was dctected by flow cytometry; the expression level of apoptosis-related protein Cyclin D1, Bcl-2, procaspase-3 and PI3K/ Akt signaling pathway protein phosphorylated-Akt （ p-Akt）, phosphorylated-TOR （ p-mTOR）, phosphorylated-PTOS6K （p-P70S6K） phosphorylated-Akt （p-Akt） in Jeko-1 cells were determined by Western blot. The results showed that the growth of Jeko-1 cell line was inhibited by LY294002. The apoptosis rates of Jeko-1 cells treated with 0,5,10 and 20 ixmo1/LofLY294002 for 24 hours were （3.25 ± 1.27）%, （11.34 ± 2.35）%, （22.81 ±2.74）%, （43.61 ± 3.48） % respectively, the difference between them was statistically significant （P 〈 0.01 ）. Phosphorylation levels of PI3K/Akt signaling pathway protein p-AM, p-mTOR, p-PTOS6K decreased, the expression of apoptosis-related protein cyclin D1, Bcl-2, procaspase-3 was down-regulated. It is concluded that the LY294002 can inhibit Jeko-1 cell proliferation, which may be realized through down-regulating the phosphorylation level of p-Akt, p-mTOR, p-PTOS6K, inhibiting the P13k/Akt signaling pathway, and promoting the cell apoptosis.