干扰hsa_circ_0004771对顺铂耐药胃癌细胞增殖、凋亡和迁移的影响  

Effect of interfering hsa_circ_0004771 on the proliferation,apoptosis and migration of cisplatin-resistant gastric cancer cells

作　　者：王竞[1] 段志英[1] 杨明月 朱秀芳[1] 霍晓辉[1] 范红云[1] WANG jing;DUAN Zhi-ying;YANG Ming-yue;ZHU Xiu-fang;HUO Xiao-hui;FAN Hong-yun(Department of Gastroenterology,The First Hospital of Hebei Medical University,Shijiazhuang 050081,Hebei Province,China)

机构地区：[1]河北医科大学第一医院消化内科,河北石家庄050081

出　　处：《中国临床药理学杂志》2022年第14期1633-1638,共6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(20180218)。

摘　　要：目的探讨hsa_circ_0004771对顺铂(DDP)耐药胃癌细胞增殖、凋亡和迁移的影响。方法体外培养胃癌细胞HGC-27和顺铂耐药胃癌细胞HGC-27/DPP,用实时定量聚合酶链反应(RT-qPCR)法检测hsa_circ_0004771和miR^(-1)49的表达水平。用双荧光素酶报告基因实验验证HGC-27/DPP细胞中hsa_circ_0004771和miR^(-1)49的调控关系。将HGC-27/DPP细胞分为hsa_circ_0004771小干扰RNA(si-hsa_circ_0004771)组(转染si-hsa_circ_0004771)、小干扰RNA阴性对照(si-NC)组(转染si-NC)、miR^(-1)49组(转染miR^(-1)49模拟物)、模拟对照序列(miR-NC)组(转染miR-NC)、si-hsa_circ_0004771+miR^(-1)49抑制剂(anti-miR^(-1)49)组(共转染si-hsa_circ_0004771和anti-miR^(-1)49)、si-hsa_circ_0004771+抑制剂阴性对照序列(anti-miR-NC)组(共转染si-hsa_circ_0004771和anti-miR-NC),用细胞计数试剂盒-8(CCK-8)法和克隆形成实验检测细胞的增殖情况,用流式细胞术检测细胞的凋亡情况,用划痕实验检测细胞的迁移情况。结果HGC-27和HGC-27/DPP细胞中hsa_circ_0004771相对表达水平分别为1.00±0.00和3.60±0.20,miR^(-1)49相对表达水平分别为1.00±0.00和0.12±0.01,差异均有统计学意义(均P<0.05)。hsa_circ_0004771在HGC-27/DPP细胞中靶向调控miR^(-1)49。si-hsa_circ_0004771组、si-NC组、miR^(-1)49组、miR-NC组、si-hsa_circ_0004771+anti-miR^(-1)49组和si-hsa_circ_0004771+anti-miR-NC组的细胞抑制率分别为(54.03±2.38)%,0,(47.27±3.18)%,0,(17.65±1.25)%和(54.52±2.20)%,集落形成数分别为(44.67±1.25),(104.00±2.94),(54.33±2.49),(104.00±3.56),(91.33±3.86)和(45.00±1.63)个,凋亡率分别为(22.35±1.56)%,(8.31±0.46)%,(19.52±1.03)%,(8.10±0.44)%,(22.35±1.50)%和(13.48±0.80)%,划痕愈合率分别为(27.14±1.21)%,(64.82±2.41)%,(34.08±1.18)%,(65.02±2.43)%,(50.67±2.92)%和(27.15±1.35)%。si-hsa_circ_0004771组的上述指标与si-NC组比较、miR^(-1)49组的上述指标与miR-NC组比较、si-hsa_circ_0004771+anti-miR^(-1)49组的上述指标与si-hsa_circ_0004771+anti-miR-NCObjective To explore the effect and mechanism of hsa_circ_0004771 on the proliferation,apoptosis and migration of cisplatin(DDP)resistant gastric cancer cells.Methods The gastric cancer cells HGC-27 and cisplatin-resistant gastric cancer cells HGC-27/DPP were cultured in vitro,and the expressions of hsa_circ_0004771 and microRNA^(-1)49(miR^(-1)49)in the cells were detected by real-time quantitative polymerase chain reaction(RT-qPCR).The dual luciferase reporter gene experiment was used to verify the regulatory relationship between hsa_circ_0004771 and miR^(-1)49 in HGC-27/DPP cells.HGC-27/DPP cells were divided into hsa_circ_0004771 small interfering RNA(si-hsa_circ_0004771)group(transfected with si-hsa_circ_0004771),small interfering RNA negative control(si-NC)group(transfected with si-NC),miR^(-1)49 group(transfected with miR^(-1)49 mimics),mock control sequence(miR-NC)group(transfected with miR-NC),si-hsa_circ_0004771+miR^(-1)49 inhibitor(anti-miR^(-1)49)group(co-transfected with si-hsa_circ_0004771 and anti-miR^(-1)49),and si-hsa_circ_0004771+inhibitor negative control sequence(anti-miR-NC)group(co-transfected with si-hsa_circ_0004771 and anti-miR-NC).Cell counting kit-8(CCK-8)and clone formation test were used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.Scratch test was used to detect cell migration.Results The expression levels of hsa_circ_0004771 in HGC-27 cells and HGC-27/DPP cells were 1.00±0.00 and 3.60±0.20;the expression levels of miR^(-1)49 were 1.00±0.00 and 0.12±0.01,respectively.The expression differences of hsa_circ_0004771 and miR^(-1)49 in the two cells were statistically significant(all P<0.05).hsa_circ_0004771 could target and negatively regulate the expression of miR^(-1)49.Inhibition rates of HGC-27/DPP cells in si-hsa_circ_0004771,si-NC,miR^(-1)49,miR-NC,si-hsa_circ_0004771+anti-miR^(-1)49 and si-hsa_circ_0004771+anti-miR-NC groups were(54.03±2.38)%,0,(47.27±3.18)%,0,(17.65±1.25)%and(54.52±2.20)%;the number of colonies formed was 44.67±1.25,104.00

关 键 词：胃癌 顺铂耐药 微小RNA-149 细胞增殖 凋亡 迁移 

分 类 号：R735.2[医药卫生—肿瘤]