CircCSNK1G1调节miR-381-3p/BRD4轴对喉癌细胞恶性生物学行为的影响

目的:探讨CircCSNK1G1调节miR-381-3p/溴结构域蛋白4(BRD4)轴对喉癌(laryngeal cancer, LC)细胞恶性生物学行为的影响。方法:实时荧光定量PCR(qRT-PCR)、western blot分别检测60例LC患者癌组织、癌旁组织及喉上皮细胞NP69及人LC细胞Hep-2、Tu212、M4E中CircCSNK1G1、miR-381-3p、BRD4蛋白表达；将人喉癌Hep-2细胞分为：control组(正常培养)、si-NC组(转染si-NC)、si-CircCSNK1G1组(转染si-CircCSNK1G1)、si-CircCSNK1G1+inhibitor-NC组(si-CircCSNK1G1和inhibitor-NC共转染)、si-CircCSNK1G1+miR-381-3p inhibitor组(si-CircCSNK1G1和miR-381-3p inhibitor共转染);RT-qPCR检测Hep-2细胞中CircCSNK1G1、miR-381-3p的表达水平；MTT法检测Hep-2细胞增殖；划痕实验检测Hep-2细胞迁移；Transwel...

Impact of CircCSNK1G1 on the malignant biological behavior of laryngeal cancer cells by regulating the miR-381-3p/bromine domain protein 4(BRD4) axis

Objective:To investigate the impact of CircCSNK1G1 on the malignant biological behavior of laryngeal cancer(LC) cells by regulating the miR-381-3p/bromine domain protein 4(BRD4) axis.Methods:Real-time fluorescence quantitative PCR(qRT-PCR) and western blot were used to detect the expression of CircCSNK1G1,miR-381-3p and BRD4 in cancer tissues,adjacent tissues and laryngeal epithelial cells NP69 and human LC cells Hep-2,Tu212 and M4E of 60 LC patients.Hep-2 cells were divided into control group(normal culture),si-NC group(transfected with si-NC),si-CircCSNK1G1 group(transfected with si-CircCSNK1G1),si-CircCSNK1G1+inhibitor NC group(co transfected with si-CircCSNK1G1 and inhibitor NC),and si-CircCSNK1G1+miR-381-3p inhibitor group(co transfected with si-CircCSNK1G1 and miR-381-3p inhibitor).The expression levels of CircCSNK1G1 and miR-381-3p in Hep-2 cells were detected by RT-qPCR.The proliferation of Hep-2 cells was detected by MTT assay.Hep-2 cell migration was detected by scratch test.Transwell chamber was used to detect Hep-2 cell invasion.Hep-2 cell apoptosis was detected by flow cytometry.Western blot was used to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),Bcl-2 associated X protein(Bax),anti apoptosis factor B cell lymphoblastoma 2(Bcl-2) and BRD4 protein.Double luciferase reporter gene experiment was applied to verify the relationship between CircCSNK1G1 and miR-381-3p,miR-381-3p and BRD4 respectively.Results:In LC tissues and LC cells,CircCSNK1G1 and BRD4 proteins were highly expressed,and miR-381-3 p was lowly expressed.In Hep-2 cells,CircCSNK1G1 and BRD4 protein levels were the highest,and miR-381-3 p expression was the lowest(P＜0.05).Therefore,Hep-2 cells were selected for transfection.Compared with control group and si-NC group,the expression of CircCSNK1G1,OD490 value,scratch healing rate,invasion number,PCNA,MMP-2,Bcl-2,BRD4 proteins in Hep-2 cells of si-CircCSNK1G1 group decreased obviously,the miR-381-3p,apoptosis rate and Bax protein expression increased obviously(P＜0.05).Compared with the si-CircCSNK1G1 group and the si-CircCSNK1G1+inhibitor NC group,the OD490 value,scratch healing rate,invasion number,expression of PCNA,MMP-2,Bcl-2,BRD4 protein in Hep-2 cells of the si-CircCSNK1G1+miR-381-3p inhibitor group increased obviously,the miR-381-3p,apoptosis rate and Bax protein expression decreased obviously(P＜0.05).CircCSNK1G1 targeted miR-381-3p expression,and miR-381-3p targeted BRD4 expression negatively.Conclusion:Knocking-down CircCSNK1G1 may down regulate BRD4 expression by targeting miR-381-3p,thereby inhibiting proliferation,migration,invasion of Hep-2 cells and promoting apoptosis.