硼替佐米对骨髓瘤细胞株RPMI8226增殖和凋亡影响的研究

摘要

Objective To explore the effects of bortezomib on the proliferation and apoptosis of multiple myeloma (MM)cell line RPMI8226.Method 20,40,60,80 and 100 nmol/L of bortezomib were used to treat the MM cell line RPMI8226 as 5 treatment groups,respectively,and blank cell line not containing bortezomib was set as control.MTT as-say was utilized to detect the inhibition of bortezomib on RPMI8226,Annexin V-FITC/PI flow cytometry was applied to detect cell apoptosis rate,besides,the mRNA and protein expression of Wnt/β-catenin signaling pathway related protein β-catenin,c-myc and PI3K/AKT signaling pathway related protein Bcl-2 and Bax were determined by RT-PCR and West-ern Blot.Result Bortezomib at any concentration could inhibit the proliferation of RPMI8226 cells,presenting a dose-dependent effect;after treatment with the different bortezomib concentrations of 20,40,60,80 and 100 nmol/L,the apop-tosis rate of RPMI8226 cells were(11.80±0.56)%,(19.45±1.25)%,(36.82±2.26)%,(43.56±3.50)% and(56.62±5.02)%,re-spectively,and all were higher than that in the control group at(8.02±0.45)%,with statistically significant difference ob-served(P<0.05);the mRNA and protein expression of c-myc,β-catenin and Bcl-2 in all bortezomib-treated groups were lower than that in control group,and higher inhibition was observed as the concentration of bortezomib increased;while the Bax mRNA and protein expression was higher than that in control group,and increased along with the elevation of bortezomib concentration.Conclusion Bortezomib can inhibit the proliferation of the MM cells,promoting its apopto-sis,which may be related to the inhibition of Wnt/β-catenin signaling pathway and PI3K/AKT signal pathway.%目的 探讨硼替佐米对骨髓瘤细胞株RPMI8226增殖和凋亡的影响.方法 采用20、40、60、80和100 nmol/L的硼替佐米处理骨髓瘤细胞株RPMI8226(处理组),以不含硼替佐米的细胞作为对照(对照组).采用MTT法检测硼替佐米对RPMI8226细胞的增殖抑制作用,采用Annexin V-FITC/PI流式细胞术检测细胞凋亡率,采用RT-PCR和Western Blot技术检测细胞Wnt/β-catenin信号通路相关蛋白β-catenin、c-myc及PI3K/AKT信号通路相关蛋白Bcl-2、Bax的mRNA和蛋白表达水平.结果 不同浓度的硼替佐米均可抑制RPMI8226细胞的增殖,且呈剂量效应;20、40、60、80和100 nmol/L硼替佐米处理组的RPMI8226细胞凋亡率分别为(11.80±0.56)%、(19.45±1.25)%、(36.82±2.26)%、(43.56±3.50)%和(56.62±5.02)%,均高于对照组的(8.02±0.45)%,差异均有统计学意义(P﹤0.05).处理组c-myc、β-catenin及Bcl-2的mRNA和蛋白表达水平均低于对照组,且随硼替佐米浓度的增加而下降;而Bax的mRNA和蛋白表达水平均高于对照组,且随硼替佐米浓度的增加而升高.结论 硼替佐米可抑制多发性骨髓瘤细胞的增殖,促进其凋亡,作用机制可能与调控Wnt/β-catenin信号通路和PI3K/AKT信号通路有关.