花旗松素调节MCP-1/CCR2轴对急性淋巴细胞白血病细胞恶性进展的影响

Impact of Taxifolin on the malignant progression of acute lymphoblastic leukemia cells by regulating the MCP-1/CCR2 axis

目的:研究花旗松素(Taxifolin)对急性淋巴细胞白血病(ALL)细胞恶性进展的影响及单核细胞趋化蛋白-1(CCL2)/CC趋化因子受体2(CCR2)轴发挥的作用。方法:体外培养ALL-T淋巴细胞CCRF-CEM;CCK-8法检测不同浓度Taxifolin对CCRF-CEM细胞活力影响,筛选本实验用Taxifolin浓度;将CCRF-CEM细胞分为control组、Taxifolin-L组、Taxifolin-M组、Taxifolin-H组、Taxifolin+rCCL2组;EDU法检测各组CCRF-CEM细胞增殖;流式细胞术检测各组CCRF-CEM细胞凋亡;Transwell检测各组CCRF-CEM细胞侵袭能力;平板克隆形成实验检测各组CCRF-CEM细胞集落形成能力;RT-PCR检测各组CCRF-CEM细胞CCL2、CCR2基因表达水平;Western blot检测各组CCRF-CEM细胞CCL2、CCR2、Bcl-2、Bax、Caspase-3蛋白表达水平。结果:选择25、50、100μmol/L三个Taxifolin浓度进行后续实验;与control组相比,Taxifolin-L、M、H组CCRF-CEM细胞凋亡率、Bax、Caspase-3蛋白表达水平显著升高,CCRF-CEM细胞EDU阳性细胞率、侵袭细胞数量、细胞克隆集落数、CCL2、CCR2基因表达水平、CCL2、CCR2、Bcl-2蛋白表达水平均显著性降低,且呈剂量依赖性(P<0.05);与Taxifolin-H组相比,Taxifolin+rCCL2组CCRF-CEM细胞凋亡率、Bax、Caspase-3蛋白表达水平显著性降低,CCRF-CEM细胞EDU阳性细胞率、侵袭细胞数量、细胞克隆集落数、CCL2、CCR2基因表达水平、CCL2、CCR2、Bcl-2蛋白表达水平均显著性升高(P<0.05)。结论:Taxifolin可能通过抑制CCL2/CCR2轴,从而抑制CCRF-CEM细胞恶性进展。

Objective:To investigate the impact of Taxifolin on the malignant progression of acute lymphoblastic leukemia(ALL) cells and the role of monocyte chemotactic protein-1(CCL2)/CC chemokine receptor 2(CCR2) axis.Methods:ALL-T lymphocytes CCRF-CEM were cultured in vitro.CCK-8 method was applied to detect the effects of different concentrations of Taxifolin on the viability of CCRF-CEM cells,and the concentration of Taxifolin was selected for this experiment.CCRF-CEM cells were divided into control group,Taxifolin-L group,Taxifolin-M group,Taxifolin-H group,and Taxifolin + r CCL2 group.EDU method was applied to detect the proliferation of CCRF-CEM cells in each group.Flow cytometry was applied to detect apoptosis of CCRF-CEM cells in each group.Transwell was applied to detect the invasiveness of CCRF-CEM cells in each group.Plate cloning experiment was applied to detect the colony forming ability of CCRF-CEM cells in each group.RT-PCR was applied to detect the expression levels of CCL2 and CCR2 genes in CCRF-CEM cells in each group and Western blot was applied to detect the expression levels of CCL2,CCR2,Bcl-2,Bax,and Caspase-3 proteins in CCRF-CEM cells in each group.Results:25,50 and 100 μmol/L Taxifolin were selected for subsequent experiments.Compared with the control group,the apoptosis rate of CCRF-CEM cells,and the protein expression levels of Bax and Caspase-3 in Taxifolin-L,M,and H groups were obviously higher,the EDU positive cell rate of CCRF-CEM cells,number of invading cells,number of cell clones,expression levels of CCL2 and CCR2 genes,and expression levels of CCL2,CCR2,and Bcl-2 proteins in CCRF-CEM cells were obviously lower,and it was dose-dependent( P < 0.05).Compared with the Taxifolin-H group,the apoptosis rate of CCRF-CEM cells,and the protein expression levels of Bax and Caspase-3 in Taxifolin + r CCL2 groups were obviously lower,the EDU positive cell rate of CCRF-CEM cells,number of invading cells,number of cell clones,expression levels of CCL2 and CCR2 genes,and expression levels of CCL2,CCR2,and Bcl-2 proteins were obviously higher( P < 0.05).Conclusion:Taxifolin may inhibit the malignant progression of CCRF-CEM cells by inhibiting the CCL2/CCR2 axis.