CircBACH1调节miR-101-3p/KPNA2轴对骨肉瘤细胞增殖、凋亡和侵袭的影响  

Impacts of circBACH1 on proliferation,apoptosis,and invasion of osteosarcoma cells by regulating the miR-101-3p/KPNA2 axis

作　　者：张立喜[1] 杨健 史博 ZHANG Lixi;YANG Jian;SHI Bo(Department of Orthopedics,Tangshan Fengnan District Hospital,Hebei Tangshan 063300,China;Department of Orthopedics,Cangzhou Central Hospital,Hebei Cangzhou 061000,China;Department of Orthopedics,Cangzhou People's Hospital,Hebei Cangzhou 061000,China)

机构地区：[1]唐山市丰南区医院骨科,河北唐山063300 [2]沧州市中心医院骨科,河北沧州061000 [3]沧州市人民医院骨科,河北沧州061000

出　　处：《现代肿瘤医学》2024年第22期4254-4261,共8页Journal of Modern Oncology

基　　金：河北省医学科学研究课题计划项目(编号:20200332)。

摘　　要：目的:探究环状RNA BACH1(circBACH1)靶向微小RNA-101-3p(miR-101-3p)/核转运蛋白α2(KPNA2)轴对骨肉瘤(OS)细胞增殖、凋亡和侵袭的影响。方法:qRT-PCR法分析OS组织中circBACH1和miR-101-3p表达水平。双荧光素酶检测circBACH1和miR-101-3p及KPNA2和miR-101-3p的靶向关系。在OS细胞MG-63中转染相应质粒记为si-NC组、si-circBACH1组、si-circBACH1+anti-NC组、si-circBACH1+anti-miR-101-3p组、miR-NC组、miR-101-3p mimics组、miR-101-3p mimics+pcDNA组、miR-101-3p mimics+KPNA2组。平板克隆检测细胞增殖;Annexin V-FITC/PI法检测细胞凋亡;Transwell实验检测细胞侵袭能力。Western blot法检测细胞周期蛋白D1(Cyclin D1)、裂解的胱天蛋白酶-3(C-caspase-3)、上皮细胞钙黏蛋白(E-cadherin)、神经性钙黏附蛋白(N-cadherin)和波形蛋白(Vimentin)的蛋白表达变化。小鼠移植瘤实验验证circBACH1对OS肿瘤生长及miR-101-3p/KPNA2的影响。结果:在OS组织中,circBACH1表达上调,miR-101-3p表达下调(P<0.05)。抑制circBACH1表达,可显著抑制OS细胞增殖与侵袭,诱导凋亡(P<0.05)。抑制miR-101-3p表达可促进OS细胞增殖与侵袭,抑制细胞凋亡(P<0.05)。miR-101-3p过表达可下调KPNA2表达,抑制OS细胞增殖与侵袭,诱导凋亡(P<0.05)。KPNA2过表达可以逆转过表达miR-101-3p对OS细胞增殖与侵袭的抑制作用(P<0.05)。小鼠移植瘤实验结果显示,抑制circBACH1表达,移植瘤体积及质量降低,miR-101-3p表达水平升高,KPNA2表达水平降低(P<0.05)。结论:circBACH1在OS组织中高表达,抑制circBACH1表达,可抑制OS细胞增殖与侵袭,诱导凋亡,其与调节miR-101-3p/KPNA2轴有关。Objective:To explore the impacts of circular RNA BACH1(circBACH1)on proliferation,apoptosis,and invasion of osteosarcoma(OS)cells by targeting microRNA-101-3p(miR-101-3p)/nuclear transporter protein karyopherinα2(KPNA2)axis.Methods:The qRT-PCR method was applied to analyze the expression levels of circBACH1 and miR-101-3p in OS tissues.Dual Luciferase was applied to detect the targeting relationship between circBACH1 and miR-101-3p,and between KPNA2 and miR-101-3p.The corresponding plasmid transfected in OS cells MG-63 was labeled si-NC group,si-circBACH1 group,si-circBACH1+anti-NC group,si-circBACH1+anti-miR-101-3p group,miR-NC group,miR-101-3p mimics group,and miR-101-3p mimics+pcDNA group,miR-101-3p mimics+KPNA2 group.Plate cloning was applied to detect cell proliferation.Annexin V-FITC/PI method was applied to detect cell apoptosis.Transwell experiment was applied to detect cell invasion ability.Western blot was applied to detect the protein expression changes of cyclin D1,C-caspase-3,E-cadherin,N-cadherin and Vimentin.Mouse transplantation tumor experiment was applied to verify the effect of circBACH1 on OS tumor growth and miR-101-3p/KPNA2.Results:In OS tissues,circBACH1 expression was up-regulated and miR-101-3p expression was down-regulated(P<0.05).Inhibiting the expression of circBACH1 was able to obviously inhibit the proliferation and invasion of OS cells,and induce apoptosis(P<0.05).Inhibition of miR-101-3p expression can promote the proliferation and invasion of OS cells and inhibit cell apoptosis(P<0.05).Overexpression of miR-101-3p can down-regulate the expression of KPNA2,inhibit the proliferation and invasion of OS cells,and induce apoptosis(P<0.05).Overexpression of KPNA2 was able to reverse the inhibitory effect of overexpression of miR-101-3p on OS cell proliferation and invasion(P<0.05).The results of the mouse transplantation tumor experiment showed that inhibiting the expression of circBACH1 resulted in a decrease in the volume and quality of the transplanted tumor,an increase in the expre

关 键 词：环状RNA BACH1 微小RNA-101-3p 核转运蛋白karyopherinα2 骨肉瘤 增殖 凋亡 侵袭 

分 类 号：R738.1[医药卫生—肿瘤]