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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 and RAD001 on the proliferation and apoptosis of diffuse large B cells SUDHL-4 and DB cells. Methods: Cell proliferation was detected by MTT method. Cell cycle and apoptosis were detected by flow cytometry. The expression of mTOR, the upstream and downstream of the signaling pathway, And apoptosis-related molecules. RESULTS: RAD001 inhibited the proliferation of SUDHL-4 and DB cells in a dose-dependent manner. Rituximab promoted the apoptosis of SUDHL-4 and DB cells. RAD001 induced the apoptosis of SUDHL-4 and DB cells The effect was not obvious, but the cell cycle arrest in G0 / G1 phase. Western blotting showed that the combination therapy could decrease the activity of downstream target protein, down-regulate cell cycle related protein and anti-apoptotic protein, and up-regulate apoptosis-related protein. Conclusions: The combination of rituximab and RAD001 can synergistically inhibit the growth of SUDHL-4 and DB cells in diffuse large B cell line, arrest the cell cycle in G0 / G1 phase and promote the apoptosis of SUDHL-4 and DB cells.