摘要
目的探讨AURKA对多发性骨髓瘤(MM)细胞增殖、凋亡、周期改变及对药物敏感性的影响,并分析内在机制。方法人多发性骨髓瘤细胞株RPMI8226和U266分别转染smart silencer NC(ssi-NC)和smart silencer AURKA(ssi-AURKA),分为ssi-NC组、ssi-AURKA组和空白组,采用实时荧光定量聚合酶链式反应(qRT-PCR)和蛋白质印迹实验检测3组细胞AURKA mRNA和AURKA蛋白相对表达量,细胞计数试剂盒8(CCK8)检测其细胞增殖活性,流式细胞术检测ssi-NC组、ssi-AURKA组细胞周期变化和凋亡率,CCK 8法检测其对硼替佐米和地塞米松的药物敏感性,蛋白质印迹实验检测Wnt信号通路相关因子及上皮细胞-间充质转化(EMT)相关蛋白表达。结果RPMI8226细胞ssi-AURKA组蛋白相对表达量为0.58±0.09,低于空白组(1.00±0.10)和ssi-NC组(1.00±0.09),F=20.84,P<0.001;U266细胞ssi-AURKA组蛋白相对表达量为0.57±0.07,低于空白组(1.00±0.04)和ssi-NC组(1.07±0.08),F=46.19,P<0.001。细胞增殖活性实验结果显示RPMI8226细胞空白组、ssi-NC组、ssi-AURKA组相对活性差异有统计学意义,F_(时间)=3966.12,P_(时间)<0.001;F_(组别)=363.15,P_(组别)<0.001;F_(时间×组别)=48.03,P_(时间×组别)<0.001。U266细胞空白组、ssi-NC组、ssi-AURKA组相对活性差异有统计学意义,F_(时间)=2452.32,P_(时间)<0.001;F_(组别)=236.91,P_(组别)<0.001;F_(时间×组别)=33.47,P_(时间×组别)<0.001。RPMI8226细胞和U266细胞中ssi-AURKA组S期均增加,t值分别为12.28和14.64,均P<0.001;G_(0)/G_(1)期均降低,t值分别为23.33和22.98,均P<0.001。RPMI8226细胞(t=16.23,P<0.001)和U266细胞(t=18.44,P<0.001)中,与ssi-NC组比较,ssi-AURKA组细胞总凋亡率增加。药物敏感性实验结果显示,RPMI8226细胞ssi-NC组、ssi-AURKA组、ssi-NC+B+D组、ssi-AURKA+B+D组细胞活性差异有统计学意义,F_(时间)=618.82,P_(时间)<0.001;F_(组别)=1373.51,P_(组别)<0.001;F_(时间×组别)=228.70,P_(时间×组别)<0.001。U266细胞ssi-NC组、ssi-AURKA组、ssi-NC
Objective To explore the effects of AURKA on proliferation,apoptosis,cycle changes and drug sensitivity of multiple myeloma(MM)cells,and analyze the underlying mechanisms.Method Human multiple myeloma cell lines RPMI8226 and U266 were transfected with smart silent NC(ssi-NC)and smart silent AURKA(ssi-AURKA)respectively.They were divided into ssi-NC group,ssi-AURKA group and blank group.Real time fluorescence quantitative polymerase chain reaction(qRT-PCR)and western blotting were used to detect the relative expression levels of AURKA mRNA and AURKA protein in the three groups of cells.The cell proliferation activity was detected by CCK8 method,and the cell cycle changes and apoptosis rate were detected by flow cytometry in the ssi-NC group and ssi-AURKA group.The drug sensitivity to bortezomib and dexamethasone was evaluated by CCK 8 assay.Western blotting was performed to detect the expression of Wnt signaling pathway related factors and epithelial mesenchymal transition(EMT)related proteins.Results The relative expression level of ssi-AURKA histone in RPMI8226 cells was 0.58±0.09,lower than that of the blank group(1.00±0.10)and ssi-NC group(1.00±0.09),F=20.84,P<0.001;The relative expression level of ssi-AURKA histone in U266 cells was 0.57±0.07,lower than that of the blank group(1.00±0.04)and ssi-NC group(1.07±0.08),F=46.19,P<0.001.The results of the cell proliferation activity experiment showed that there was a statistically significant difference in the relative activity of RPMI8226 cells among the blank group,ssi-NC group and ssi-AURKA group,F_(time)=3966.12,P_(time)<0.001;F_(group)=363.15,P_(group)<0.001;F_(time×group)=48.03,P_(time×group)<0.001.There was a statistically significant difference in relative cell activity among the U266 cell blank group,ssi-NC group and ssi-AURKA group,with F_(time)=2452.32 and P_(time)<0.001;F_(group)=236.91,P_(group)<0.001;F_(time×group)=33.47,P_(time×group)<0.001.The S phase of the ssi-AURKA group in RPMI8226 cells and U266 cells were increased,with t-values of 12
作者
刘红春
黄睿
许腾
黄国虹
LIU Hongchun;HUANG Rui;XU Teng;HUANG Guohong(Clinical Laboratory Center,Xinjiang Uygur Autonomous Region People's Hospital,Urumqi,Xinjiang 830002,China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2024年第14期865-873,共9页
Chinese Journal of Cancer Prevention and Treatment
基金
新疆维吾尔自治区自然科学基金面上项目(2021D01C135)
新疆维吾尔自治区人民医院院内项目(20200209)
