STAT1在多发性骨髓瘤增殖及细胞黏附介导耐药中的作用

Role of STAT1 in the proliferation and cell adhesion-mediated drug resistance in multiple myeloma

目的分析信号转导与转录激活因子1(signal transducer and activator of transcription 1,STAT1)对多发性骨髓瘤增殖及细胞黏附介导的耐药(cell adhesion-mediated drug resistance,CAM-DR)影响。方法H929及MM.1S细胞感染STAT1过表达慢病毒(Lv-STAT1)或对照慢病毒(Lv-Ctrl),CCK-8法分析过表达STAT1对H929及MM.1S细胞增殖的影响;CCK-8实验、台盼蓝拒染实验分析过表达STAT1对CAM-DR的影响;Western blot分析过表达STAT1对IRF2、BCL-2、Bax、Cleaved Caspase-3表达的影响。结果与对照组相比,过表达STAT1可显著增强H929及MM.1S细胞的增殖能力(H929细胞48 h:t=5.78,P=0.004;H929细胞72 h:t=4.97,P=0.008;MM.1S细胞48 h:t=3.27,P=0.03;MM.1S细胞72 h:t=5.01,P=0.007)。与对照组相比,过表达STAT1可显著降低H929及MM.1S细胞对多柔比星的敏感性。与对照组相比,过表达STAT1可显著降低悬浮及黏附培养状态下多柔比星诱导的细胞死亡(H929悬浮:t=8.43,P=0.001;H929黏附:t=4.63,P=0.009;MM.1S悬浮:t=3.94,P=0.017;MM.1S黏附:t=3.91,P=0.017)。过表达STAT1可增强H929以及MM.1S细胞中IRF2、BCL-2的蛋白表达水平,并抑制Bax、Cleaved Caspase-3的蛋白表达水平。结论STAT1异常活化可促进多发性骨髓瘤细胞的增殖和CAM-DR,其可能通过激活STAT1-IRF2信号通路发挥作用。

Purpose To investigate the effect of signal transducer and activator of transcription 1(STAT1)on cell proliferation and cell adhesion-mediated drug resistance(CAM-DR)phenotype in multiple myeloma.Methods H929 and MM.1S cells were infected with lentivirus expressing STAT1(Lv-STAT1)or lentivirus of empty vector(Lv-Ctrl).Then,CCK-8 assay was utilized to assess the effect of STAT1 overexpression on the proliferation of H929 and MM.1S cells.CCK-8 assay and typan blue dye exclusion method were used to determine the effect of STAT1 overexpression on CAM-DR phenotype.Western blot assay was used to detect the effect of STAT1 overexpression on IRF2,BCL-2,Bax,and Cleaved Caspase-3 expression.Results Overexpression of STAT1 significantly enhanced the proliferation of H929 and MM.1S cells compared with the control group(H929 cells 48 h after infection:t=5.78,P=0.004.H929 cells 72 h after infection:t=4.97,P=0.008.MM.1S cells 48 h after infection:t=3.27,P=0.03,MM.1S cells 72 h after infection:t=5.01,P=0.007).Furthermore,ectopic expression of STAT1 decreased sensitivity of H929 and MM.1S cells to doxorubicin.Intriguingly,overexpression of STAT1 remarkably decreased doxorubicin-induced cell death in both suspension and fibronectin(FN)-adherent cells compared with the control group(H929 cells kept in suspension:t=8.43,P=0.001,H929 cells adhered to FN:t=4.63,P=0.009,MM.1S cells kept in suspension:t=3.94,P=0.017,MM.1S cells adhered to FN:t=3.91,P=0.017).Moreover,ectopic expression of STAT1 remarkably increased IRF2 and BCL-2 expression,but decreased Bax and Cleaved Caspase-3 expression in both H929 and MM.1S cells.Conclusion The abnormal activation of STAT1 promotes proliferation and CAM-DR in multiple myeloma cells,possibly through activation of the STAT1-IRF2 signaling pathway.