长链非编码RNA ZFAS1通过miR-193b-3p影响胶质瘤顺铂敏感性研究  

Influence of lncRNA ZFAS1 on cisplatin sensitivity in glioma via miR-193b-3p regulation

作　　者：张慧[1] 郑款云 齐丽娜 薛子娇 ZHANG Hui;ZHENG Kuan-yun;QI Li-na;XUE Zi-jiao(Clinical Laboratory,Tangshan City Workers'Hospital,Tangshan 063000,Hebei Province,China;College of Nursing and Rehabilitation,North China University of Science and Technology,Tangshan 063000,1Hebei Province,China;Rehabilitation Medicine Department,Affiliated Hospital of North China Universityof Science and Technology,Tangshan 063000,Hebei Province,China)

机构地区：[1]唐山市工人医院检验科,河北唐山063000 [2]华北理工大学护理与康复学院,河北唐山063000 [3]华北理工大学附属医院康复医学科,河北唐山063000

出　　处：《中国临床药理学杂志》2024年第12期1774-1778,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河北省医学科学研究重点课题计划基金资助项目(20240819)。

摘　　要：目的 研究长链非编码RNA锌指结构反义转录本1(ZFAS1)在胶质瘤细胞中对顺铂敏感性的作用及其潜在机制。方法 通过实时荧光定量聚合酶链反应(qRT-PCR)实验分析敲低ZFAS1对胶质瘤细胞顺铂敏感性,分为sh-NC组(转染sh-NC慢病毒质粒)、sh#1组(转染sh-ZFAS1-1慢病毒质粒)、sh#2组(转染sh-ZFAS1-2慢病毒质粒)。双荧光素酶实验验证ZFAS1与miR-193b-3p之间的相互作用,分为ZFAS1-WT+NC inhibitor组(转染ZFAS1野生型质粒和NC inhibitor)、ZFAS1-WT+miR-193b-3p inhibitor组(转染ZFAS1野生型质粒和miR-193b-3p inhibitor)、ZFAS1-Mut+NC inhibitor组(转染ZFAS1突变型质粒和NC inhibitor)、ZFAS1-Mut+miR-193b-3p inhibitor组(转染ZFAS1突变型质粒和miR-193b-3p inhibitor)。用细胞计数试剂盒-8(CCK-8)和原位末端标记(TUNEL)实验分析ZFAS1/miR-193b-3p影响胶质瘤细胞对顺铂敏感性,分为空白对照组(0μg·mL^(-1)顺铂处理U251细胞)、0.5μg·mL^(-1)顺铂+sh-NC+NC inhibitor组(0.5μg·mL^(-1)顺铂处理共转染sh-NC慢病毒质粒和NC inhibitor的U251细胞)、0.5μg·mL^(-1)顺铂+sh#1+NC inhibitor组(0.5μg·mL^(-1)顺铂处理共转染sh-NC慢病毒质粒和NC inhibitor的U251细胞)和0.5μg·mL^(-1)顺铂+sh#1+miR-193b-3p inhibitor组(0.5μg·mL^(-1)顺铂处理共转染sh-ZFAS1-1慢病毒质粒和miR-193b-3p inhibitor的U251细胞)。结果 sh-NC组、sh#1组和sh#2组的ZFAS1的表达水平分别为1.00±0.17、0.48±0.06和0.68±0.08。ZFAS1-WT+NC inhibitor组、ZFAS1-WT+miR-193b-3p inhibitor组、ZFAS1-Mut+NC inhibitor组、ZFAS1-Mut+miR-193b-3p inhibitor组的荧光活性分别为1.00±0.10、1.45±0.11、1.02±0.09和0.97±0.13。空白对照组、0.5μg·mL^(-1)顺铂+sh-NC+NC inhibitor组、0.5μg·mL^(-1)顺铂+sh#1+NC inhibitor组、0.5μg·mL^(-1)顺铂+sh#1+miR-193b-3p inhibitor组的72 h增殖率分别为(100.00±14.13)%、(96.62±9.82)%、(60.56±6.08)%和(78.64±7.22)%;72 h凋亡率分别为(9.52±1.11)%、(10.12±1.34)%、(16.08±1.52)%和(12.22±1.19)%。空白对照组与Objective To investigate the role of long non-coding RNA(Inc RNA) ZFAS1 in glioma cells ' sensitivity to cisplatin and its underlying mechanisms.Methods By analyzing the knockdown of ZFAS1 on the sensitivity of glioma cells to cisplatin using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR) experiments,and the cells were divided into sh-NC group(transfected with sh-NC lentiviral plasmid),sh#1 group(transfected with sh-ZFAS1-1 lentiviral plasmid) and sh#2 group(transfected with sh-ZFAS1-2 lentiviral plasmid).Dual luciferase experiments verified the interaction between ZFAS1 and miR-193b-3p,and the cells were divided into ZFAS1-WT+NC inhibitor group(transfected with ZFAS1 wild-type plasmid and NC inhibitor),ZFAS1-WT+miR-193b-3p inhibitor group(transfected with ZFAS1 wild-type plasmid and miR-193b-3p inhibitor),ZFAS1-Mut+NC inhibitor group(transfected with ZFAS1 mutant plasmid and NC inhibitor) and ZFAS1-Mut+miR-193 b-3p inhibitor group(transfected with ZFAS1 mutant plasmid and miR-193b-3p inhibitor).Cell counting kit-8(CCK-8) and terminal deoxynucleotidly transferase mediated labeling(TUNEL) experiments were used to analyze the effect of ZFAS1/miR-193b-3p on the sensitivity of glioma cells to cisplatin,and the cells were divided into blank control group(0 μg · mL^(-1) cisplatin treatment of U251 cells),0.5 μg·mL^(-1) cisplatin+sh-NC+NC inhibitor group(0.5 μg·mL^(-1) cisplatin treatment of U251 cells cotransfected with sh-NC lentiviral plasmid and NC inhibitor),0.5 μg · mL^(-1) cisplatin+sh#1+NC inhibitor group(0.5 μg·mL^(-1) cisplatin treatment of U251 cells co-transfected with sh-NC lentiviral plasmid and NC inhibitor),and 0.5 μg·mL^(-1) cisplatin+sh#1+miR-193b-3p inhibitor group(0.5 μg · mL^(-1) cisplatin treatment of U251cells co-transfected with sh-ZFAS1-1 lentiviral plasmid and miR-193b-3p inhibitor).Results The results of the experiment showed that the expression levels of ZFAS1 in the sh-NC group,sh#1 group and sh#2 group were1.00±0.17,0.48±0.06 and 0.68±0.08.The fluore

关 键 词：长链非编码RNA 锌指结构反义转录本1 微小RNA-193b-3p 胶质瘤 顺铂敏感性 

分 类 号：R97[医药卫生—药品]