转录因子EB相关自噬在多发性骨髓瘤治疗中的作用及其机制研究  

Transcription factor EB related autophagy in the treatment of multiple myeloma and its mechanism

作　　者：张志华[1] 张荣娟[1] 韩凝 栗冲 王丽红[1] 邢恩鸿[1] 谷翠红[1] 郝长来[1] Zhang Zhihua;Zhang Rongjuan;Han Ning;Li Chong;Wang Lihong;Xing Enhong;Gu Cuihong;Hao Changlai(Affiliated Hospital of Chengde Medical College,Chengde 067000,China)

机构地区：[1]承德医学院附属医院,067000

出　　处：《中华血液学杂志》2021年第5期407-414,共8页Chinese Journal of Hematology

摘　　要：目的明确硼替佐米和(或)西拉美新作用于多发性骨髓瘤(MM)细胞株后细胞增殖、转录因子EB(TFEB)核转位表达变化及自噬水平,为进一步探讨TFEB对自噬的调控机制提供依据。方法体外培养MM细胞株RPMI8226及U266,并以一定浓度的硼替佐米和西拉美新处理MM细胞,CCK-8法检测细胞增殖,实时定量PCR和Western blot法检测TFEB、自噬相关因子LC3B、Beclin1、p62、LAMP1的mRNA和蛋白相对表达量。结果随着硼替佐米浓度增加及作用时间延长,两个细胞系的增殖抑制率增高(P<0.05)。硼替佐米和西拉美新联用对上述MM细胞株的增殖有协同抑制作用(P<0.05)。空白对照组、单药组、联合用药组处理MM细胞株后,细胞质中TFEB的mRNA和蛋白相对表达量依次下降(P<0.05),细胞核中TFEB的mRNA和蛋白相对表达量依次上升(P<0.05),自噬相关因子LC3B、Beclin1、LAMP1的mRNA和蛋白相对表达量依次上升,p62的mRNA和蛋白相对表达量依次下降(P<0.05)。结论硼替佐米和西拉美新具有协同抑制MM细胞增殖作用,与其诱导MM细胞株自噬表达增强相关,发生核转位的TFEB表达亦增强。Objective To clarify the effects of bortezomib combined with or without siramesine on the proliferation of multiple myeloma cell lines,the expression changes of transcription factor EBC(TFEB)nuclear translocation and the level of autophagy,and to provide basis for further exploring the regulation mechanism of transcription factor TFEB on autophagy.Methods The multiple myeloma cell lines RPMI8226 and U266 were cultured in vitro,and the multiple myeloma cells were treated with a certain concentration of bortezomib and siramesine.The changes of cell proliferation inhibition were detected by CCK-8 method.Real time PCR and Western blot were used to detect the relative expression of TFEB,autophagy-related factor LC3B,Beclin1,p62,LAMP1 mRNA and protein.Results As the concentration of bortezomib increased and the duration of action increased,the proliferation inhibition rates of the two cell lines gradually increased(P<0.05).The combination of the two drugs has a synergistic inhibitory effect on the proliferation of the above-mentioned multiple myeloma cell lines(P<0.05).In the blank control group,single drug group,and combination drug group,the relative expression of TFEB mRNA and protein in the cytoplasm decreased sequentially(P<0.05),and the relative expression of TFEB mRNA and protein in the nucleus increased sequentially(P<0.05).The relative expression of autophagy-related factors LC3B,Beclin1,LAMP1 mRNA and protein increased sequentially,and the relative expression of p62 mRNA and protein decreased sequentially(P<0.05).Conclusion Bortezomib and siramesine can synergistically inhibit the growth of multiple myeloma cells,which is related to the increased autophagy expression in multiple myeloma cell lines and the expression of TFEB with nuclear translocation is also enhanced.

关 键 词：多发性骨髓瘤 自噬 TFEB 硼替佐米 西拉美新 

分 类 号：R733.3[医药卫生—肿瘤]