摘要
目的探讨多发性骨髓瘤(MM)细胞株U266中热休克蛋白27(HSP27)诱导硼替佐米(BTZ)耐药的机制研究。方法BTZ浓度逐步递增诱导建立耐药细胞株(U266/BTZ),细胞计数试剂盒(CCK8)法检测耐药细胞株的耐药性;实时荧光定量PCR及蛋白印迹法检测BTZ处理前后U266及U266/BTZ细胞中HSP27的表达;采用HSP27-siRNA干扰U266/BTZ细胞中HSP27表达,实时荧光定量PCR及蛋白印迹法进一步分析HSP27-siRNA干扰后对细胞凋亡以及内质网应激的影响,包括cl-Caspase3、cl-Caspase8、cl-Caspase9。结果CCK8结果显示,U266/BTZ细胞株能够对多种药物产生细胞耐药性。BTZ暴露显著上调U266以及U266/BTZ细胞中HSP27 mRNA和蛋白表达(P均<0.01);HSP27-siRNA干扰能够显著下调HSP27的表达;HSP27-siRNA干扰能够增强BTZ诱导的U266/BTZ细胞凋亡,增加活化Caspase3、Caspase8和Caspase9蛋白表达水平(P均<0.01)。结论HSP27介导的MM耐药是通过抑制内质网应激,从而减少骨髓瘤细胞凋亡导致。
ObjectiveTo investigate the mechanism of heat shock proteins 27(HSP27)induced bortezomib(BTZ)resistance in multiple myeloma(MM)cell line U266.MethodsDrug-resistant cell strain U266/BTZ was induced by treatment with increasing concentrations of BTZ,and drug resistance was detected by CCK8 method.The expression of HSP27 in U266 and U266/BTZ cells before and post BTZ treatment was detected by real-time fluorescence quantitative PCR(RT-PCR)and Western blot.Hsp27-siRNA was used to interfere with the expression of HSP27 in U266/BTZ cells,and then RT-PCR and Western blot were used to identify the effects of HSP27-siRNA interference on apoptosis and endoplasmic reticulum stress,as well as the expression of cl-Caspase-3,cl-Caspase-8,and cl-Caspase-9.ResultsCCK8 assay showed that U266/BTZ cells had reduced drug resistance to many drugs.BTZ exposure significantly upregulated the expression of HSP27 mRNA and protein in U266 and U266/BTZ cells(P<0.01).Hsp27-siRNA interference significantly down-regulated the expression of HSP27.Hsp27-siRNA interference could enhance the apoptosis of U266/BTZ cells and increase the activation of Caspase-3,Caspase-8 and Caspase-9(P<0.01).ConclusionHSP27-mediated MM resistance is caused by inhibition of endoplasmic network stress to reduce myeloma cell apoptosis.
作者
苏杰
杨小静
周雪
Su Jie;Yang Xiaojing;Zhou Xue(Department of Hematology,Nanjing Jiangbei People's Hospital,Nanjing 210048,China)
出处
《中华临床医师杂志（电子版）》
CAS
北大核心
2021年第10期779-784,共6页
Chinese Journal of Clinicians(Electronic Edition)
作者简介
通信作者:苏杰,Email:sujie004@126.com
