抑制KIF18B分子表达对人急性髓系白血病HL60细胞活性的影响及其机制探讨  

Effect of inhibiting the expression of KIF18B molecules on the viability of human acute myeloid leukemia HL60 cells and its mechanism

作　　者：庹婧 汤红霞 陈启玲 侯隽[2] 廖大忠 TUO Jing;TANG Hongxia;CHEN Qiling;HOU Jun;LIAO Dazhong(Department of Tumor Hematology,Hospital of Traditional Chinese Medicine Affiliated to Southwest Medical University,Sichuan Luzhou 646000,China;Department of Hematology,Neijiang City Hospital of Traditional Chinese Medicine,Sichuan Neijiang 641107,China)

机构地区：[1]西南医科大学附属中医医院肿瘤血液二科,四川泸州646000 [2]内江市中医医院血液科,四川内江641107

出　　处：《现代肿瘤医学》2022年第7期1147-1152,共6页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:81972138)。

摘　　要：目的:探讨抑制KIF18B分子表达后对人急性髓系白血病HL60细胞增殖、凋亡、自噬的影响及其机制。方法:选择HL60细胞随机分为3组,利用脂质体2000分别转染KIF18B-siR、NC-siR及Control组细胞,采用qRT-PCR法验证转染后HL60细胞中KIF18B的表达;CCK-8法检测HL60细胞增殖能力变化;流式细胞技术检测抑制KIF18B表达对HL60细胞凋亡能力及细胞周期的影响;通过透射电镜观察各组细胞溶酶体中自噬小体变化情况;采用Western blot检测与增殖、凋亡、自噬相关的蛋白AMPK、mTOR、P70S6K、4EBP1及ULK1的表达。结果:qRT-PCR结果显示,与Control和NC-siR组相比,KIF18B-siR组KIF18B mRNA表达水平显著下降(P<0.05);CCK-8实验结果表明,siRNA干扰KIF18B表达后,可以抑制HL60细胞增殖,有统计学差异(P<0.05);流式检测结果显示,与Control组和NC-siR组相比,抑制KIF18B表达能促使HL60细胞G_(0)/G_(1)期细胞增多,S期和G_(2)/M期细胞减少,阻滞细胞周期,促进HL60细胞大量凋亡;透射电镜观察到,与Control和NC-siR组相比,KIF18B-siR组溶酶体内分布有大量的自噬小泡;Western blot结果表明,KIF18B-siR组p-AMPK和p-ULK1表达上调,p-mTOR、p-P70S6K及p-4EBP1表达下调,结果有显著差异(P<0.05)。结论:抑制KIF18B表达可以抑制人急性髓系白血病HL60细胞增殖,促使细胞周期发生阻滞,诱导凋亡和自噬,调控机制可能与激活AMPK/mTOR通路有关。Objective:To investigate the effect and mechanism of inhibiting the expression of KIF18B on the proliferation,apoptosis and autophagy of human acute myeloid leukemia HL60 cells.Methods:HL60 cells were randomly divided into 3 groups,use liposome 2000 to transfect KIF18B-siR,NC-siR and Control respectively,and the expression of KIF18B in HL60 cells after transfection was verified by qRT-PCR.CCK-8 method was used to detect the proliferation of HL60 cells.Flow cytometry to detect the effect of inhibiting the expression of KIF18B on the apoptotic ability and cell cycle of HL60 cells.Observe the autophagosomes in each group of cells by transmission electron.Observe the changes of autophagosomes in cell lysosomes by transmission electron microscope.Western blot was used to detect the expression of AMPK,mTOR,P70S6K,4EBP1 and ULK1,which related to proliferation,apoptosis and autophagy.Results:The qRT-PCR results showed that compared with the Control and NC-siR groups,the mRNA expression level of the KIF18B-siR group was significantly decreased(P<0.05).CCK-8 experiment results showed that after siRNA interferes with the expression of KIF18B,it can inhibit the proliferation of HL60 cells,with statistical difference(P<0.05).The results of flow cytometry showed that compared with the Control group and NC-siR group,inhibiting the expression of KIF18B can increase the number of cells in the G_(0)/G_(1) phase of HL60 cells,reduce the number of cells in the S and G_(2)/M phases,block the cell cycle,and promote a large number of HL60 cell apoptosis.Transmission electron microscopy showed that compared with the Control and NC-siR groups,a large number of autophagic vesicles were distributed in the lysosomes of the KIF18B-siR group.Western blot results showed that the expressions of p-AMPK and p-ULK1 were up-regulated,while the expressions of p-mTOR,p-P70S6K and p-4EBP1 were down-regulated in the KIF18B-siR group,and the results were significantly different(P<0.05).Conclusion:Inhibiting the expression of KIF18B can inhibit the proli

关 键 词：HL60细胞 KIF18B 细胞活性 机制研究 

分 类 号：R733[医药卫生—肿瘤]