利妥昔单抗和RAD001对弥漫大B细胞株SUDHL-4和DB细胞增殖和凋亡的影响

Effect of RAD001 plus rituximab on proliferation and apoptosis of DLBCL cell line SUDHL-4 and DB cells

目的:研究利妥昔单抗、RAD001单独及联合用药对弥漫大B细胞SUDHL-4和DB细胞增殖、凋亡的影响。方法:采用四甲基偶氮唑盐比色法(MTT法)检测细胞增殖,流式细胞术检测细胞周期和细胞凋亡,蛋白印迹法检测靶蛋白mTOR及该信号通路上下游分子、细胞周期和凋亡相关分子的变化。结果:RAD001可呈剂量依赖性地抑制弥漫大B细胞系SUDHL-4和DB细胞的增殖;利妥昔单抗可促进SUDHL-4和DB细胞凋亡,RAD001对SUDHL-4和DB细胞凋亡作用不明显,但可使细胞周期阻滞在G0/G1期;蛋白印迹法显示,联合用药可使靶蛋白下游分子活性下降,下调细胞周期相关蛋白及抗凋亡蛋白,上调凋亡相关蛋白。结论:利妥昔单抗与RAD001联合用药可协同抑制弥漫大B细胞系SUDHL-4和DB细胞的生长,使细胞周期阻滞在G0/G1期,促进SUDHL-4和DB细胞的凋亡。

Objective To study the effects of rituximab,RAD001 and rituximab plus RAD001 on proliferation and apoptosis of diffuse large B-cell lymphoma cell line SUDHL-4 and DB cells.Methods MTT assay was used to assess cell proliferation.Flow cytometry was performed to examine cell cycle and apoptosis.Protein expression of mTOR signaling pathway,cell cycle and apoptosis related proteins were detected by Western blot.Result Rituximab accelerated apoptosis of SUDHL-4 and DB cells.RAD001 inhibited the proliferation of SUDHL-4 and DB cells and kept the cell in G0/G1 phase,but did not have obvious effect on apoptosis.Western blot showed that rituximab plus RAD001 could decrease the activity of downstream molecules,down-regulate cell cycle related protein,and anti-apoptotic protein up-regulate apoptotic associated protein.Conclusions Rituximab and RAD001 could function cooperatively to inhibit the proliferation and promote the apoptosis of SUDHL-4 and DB cells.