新型核苷类似物FNC对Raji细胞增殖、凋亡和Bcl-6、PRDM1、C-myc表达的影响

Effect of new nucleoside analogue FNC on proliferation,apoptosis and expressions of Bcl-6,PRDM1,C-myc in cell line Raji

目的:研究2'-脱氧-2'-氟-4'-叠氮-核苷类似物(FNC)对Raji细胞增殖的影响及其作用机制。方法:分别用0、0.035、0.350、3.500、35.000和350.000μmol/L的FNC处理Raji细胞24、48、72h后,应用MTT法检测细胞增殖情况;分别用0、0.035、0.175、0.875、4.375μmol/LFNC处理Raji细胞48h后用流式细胞术检测细胞周期及凋亡,用RT-PCR与Westernblot法分别检测细胞中Bcl-6、PRDM1、C-mycmRNA及蛋白的表达。结果:FNC可显著抑制Raji细胞的增殖,呈明显的时间、剂量依赖性(F浓度=4869.984,F时间=1785.062,F交互=126.281,P均<0.001);FNC可诱导细胞凋亡,阻滞细胞周期于G0/G1期(P<0.05);随着FNC浓度的增加,Raji细胞中Bcl-6、C-mycmRNA及蛋白的表达降低(F=65.497、68.502和59.086、78.285,P均<0.001),PRDM1mRNA及蛋白的表达升高(F=81.133、85.234,P均<0.001)。结论:FNC可能通过下调Bcl-6、C-myc和上调PRDM1的表达,抑制Raji细胞的增殖,诱导细胞凋亡。

Aim： To investigate the effects of new nucleoside analogue 2＇-deoxy-2＇-β-fluoro-4＇-azide-nucleoside analogue（ FNC） on proliferation of Raji cells and its mechanism.Methods： Raji cells were treated with 0,0.035,0.350,3.500,35.000,and 350.000 μmol/L FNC for 24,48 and 72 h,then the proliferation was determined by MTT assay.Raji cells were treated with 0,0.035,0.175,0.875,4.375 μmol/L FNC for 48 h,then the cell cycle and apoptotic rate were detected by flow cytometry,and the expression levels of Bcl-6,PRDM1 and C-myc mRNA and protein were detected by RTPCR and Western blot.Results： FNC could remarkably inhibit the proliferation of Raji cells in timeand dose-dependent manner（ Fdose=4 869.984,Ftime=1 785.062,Finteraction=126.281,P0.001） .FNC could induce cell apoptosis and arrest cell cycle in G 0/G 1 phase （ P0.05 ） .The expressions of Bcl-6 and C-myc mRNA and protein decreased （ F=65.497,68.502,59.086,78.285,P0.001） ,while the expressions of PRDM1 mRNA and protein increased with the increasing dose of FNC（ F=81.133,85.234,P0.001） .Conclusion： FNC could inhibit cell proliferation through downregulating the expressions of Bcl-6 and C-myc and up-regulating the expression of PRDM1.