ALDH1A1调控Notch1信号通路在MMSCs“干性”维持中的作用和机制研究  

Study on the role and mechanism of ALDH1A1 regulating Notch1 signaling pathway in stemness maintenance of MMSCs

作　　者：王晓丽[1] 江妤 曾玉晓[1] 刘永华[1] 章俏雷[1] WANG Xiaoli;JIANG Yu;ZENG Yuxiao;LIU Yonghua;ZHANG Qiaolei(Department of Hematology,Lishui People′s Hospital in Zhejiang Province,Lishui323000,China)

机构地区：[1]浙江省丽水市人民医院血液科,浙江丽水323000

出　　处：《中国现代医生》2022年第7期21-24,30,F0003,共6页China Modern Doctor

基　　金：浙江省基础公益研究计划项目(LGF19H080003)。

摘　　要：目的研究乙酰脱氢酶家族成员A1(ALDH1A1)调控Notch1信号通路在多发性骨髓瘤干细胞(MMSCs)“干性”维持中的作用和机制。方法选用人MMSCs CD138-B细胞和人正常干细胞QSG-7701,用Western blot法检测ALDH1A1在CD138-B细胞和QSG-7701细胞中的表达情况。将CD138-B细胞分为control组、ALDH1A1敲减组、Notch1通路抑制剂组,control组不作任何处理,ALDH1A1敲减组转染10μg ALDH1A1-shRNA重组慢病毒质粒,Notch1通路抑制剂组加入20μmol/L Notch1通路抑制剂GSI,培养48 h,检测并比较三组ALDH1A1表达量、Notch1通路相关蛋白[Notch1、发状分裂相关增强子1(Hes1)]表达量、增殖率、凋亡率及“干性”相关基因[胚胎干细胞关键蛋白(NANOG)、八聚体结合转录因子4(OCT4)]表达量。结果CD138-B细胞中ALDH1A1蛋白表达量较QSG-7701细胞明显增加(P<0.05);ALDH1A1敲减组ALDH1A1蛋白表达量较control组明显减少(P<0.05);Notch1通路抑制剂组ALDH1A1蛋白表达量与control组比较,差异无统计学意义(P>0.05)。ALDH1A1敲减组和Notch1通路抑制剂组增殖率及Notch1、Hes1蛋白和NANOG、OCT4基因表达量较control组明显减少(P<0.05),凋亡率较control组明显增多(P<0.05);Notch1通路抑制剂组增殖率、凋亡率及Notch1、Hes1蛋白和NANOG、OCT4基因表达量与ALDH1A1敲减组比较,差异无统计学意义(P>0.05)。结论ALDH1A1在MMSCs中呈高表达,ALDH1A1敲减后可通过阻抑Notch1信号通路下调MMSCs“干性”维持,抑制MMSCs增殖并诱导其凋亡。Objective To investigate the role and mechanism of aldehyde dehydrogenase family 1 member A1(ALDH1A1)in regulating the Notch1 signaling pathway in the stemness maintenance of multiple myeloma stem cells(MMSCs).Methods Human MMSCs CD138-B cells and human normal stem cells QSG-7701 were selected to detect the expression of ALDH1A1 in CD138-B cells and QSG-7701 cells by Western blot.CD138-B cells were divided into the control group,the ALDH1A1 knockdown group and the Notch1 pathway inhibitor group.The control group was not treated with any treatment.The ALDH1A1 knockdown group was transfected with 10μg ALDH1A1-shRNA recombinant lentiviral plasmid.The Notch1 pathway inhibitor group was added with 20μmol/L Notch1 pathway inhibitor GSI and cultured for 48 h.The expression levels of ALDH1A1,Notch1 pathway related proteins(Notch1,hair division associated enhancer 1[Hes1]),proliferation rate,apoptosis rate and stemness related genes(embryonic stem cell key protein[NANOG],octamer binding transcription factor 4[OCT4])were detected and compared among the three groups.Results The expression of ALDH1A1 protein in CD138-B cells was significantly higher than that in QSG-7701 cells(P<0.05).The protein expression of ALDH1A1 in the ALDH1A1 knockdown group was significantly lower than that in the control group(P<0.05).No significant difference was observed in the ALDH1A1 protein expression in the Notch1 pathway inhibitor group compared with the control group(P>0.05).The proliferation rates and the expression levels of Notch1 and Hes1 proteins and NANOG and OCT4 genes in the ALDH1A1 knockdown group and the Notch1 pathway inhibitor group were significantly decreased compared with the control group(P<0.05),and the apoptosis rates were significantly increased compared with the control group(P<0.05).No significant differences were observed in the proliferation rate,apoptosis rate,and the expressions of Notch1,Hes1,NANOG and OCT4 gene in the Notch1 pathway inhibitor group compared with the ALDH1A1 knockdown group(P>0.05).Conclusion ALDH

关 键 词：乙酰脱氢酶家族成员A1 Notch1信号通路 多发性骨髓瘤干细胞 表达水平 干性维持 细胞增殖 细胞凋亡 

分 类 号：R733.3[医药卫生—肿瘤]