circBIRC6靶向miR-449b-5p调控胃癌SUN-1细胞的增殖、细胞周期及凋亡  

circBIRC6 in regulation of the proliferation,cell cycle and apoptosis of gastric cancer SUN-1 cells by targeting miR-449b-5p

作　　者：钟碧波[1] 钟芳 寇继光[1] 李光曙 ZHONG Bi-bo;ZHONG Fang;KOU Ji-guang;LI Guang-shu(Department of Gastroenterology,Xiaogan Hospital Affiliated to Wuhan University of Science and Technology,Xiaogan,Hubei 432000,China)

机构地区：[1]武汉科技大学附属孝感医院消化内科,湖北孝感432000

出　　处：《中国临床研究》2022年第11期1516-1522,共7页Chinese Journal of Clinical Research

摘　　要：目的探讨circBIRC6对胃癌细胞增殖、细胞周期及凋亡的影响及可能的作用机制。方法收集2020年1月至5月武汉科技大学附属孝感医院收治的53例胃癌患者的癌组织及其癌旁组织标本,RT-qPCR法检测胃癌组织、癌旁组织与人胃黏膜细胞GES1、人胃癌细胞SNU-1、AGS、HS-746T中circBIRC6、miR-449b-5p的表达量;以SNU-1细胞为研究对象,随机分组为si-NC组、si-circBIRC6组、miR-NC组、miR-449b-5p组、si-circBIRC6+anti-miR-NC组、si-circBIRC6+anti-miR-449b-5p组;CCK-8法、流式细胞仪分别检测细胞增殖、细胞周期及凋亡率;双荧光素酶报告实验检测miR-449b-5p过表达对野生型载体WT-circBIRC6荧光素酶活性的影响;western blot检测cleaved caspase-3蛋白表达量。结果与癌旁组织比较,胃癌组织中circBIRC6的表达量升高(P<0.05),miR-449b-5p的表达量降低(P<0.05);与GES1细胞比较,SNU-1、AGS、HS-746T细胞中circBIRC6的表达量升高(P<0.05),miR-449b-5p的表达量降低(P<0.05)。与si-NC组比较,si-circBIRC6组细胞活力和S期细胞比例降低(P<0.05),G0/G1期细胞比例、细胞凋亡率和cleaved caspase-3蛋白水平升高(P<0.05);与miR-NC组比较,miR-449b-5p组细胞活力和S期细胞比例降低(P<0.05),G0/G1期细胞比例、细胞凋亡率和cleaved caspase-3蛋白水平升高(P<0.05)。miR-449b-5p过表达可降低WT-circBIRC6的荧光素酶活性(P<0.05)。与si-circBIRC6+anti-miR-NC组比较,si-circBIRC6+anti-miR-449b-5p组细胞活力和S期细胞比例升高(P<0.05),G0/G1期细胞比例、细胞凋亡率和cleaved caspase-3蛋白水平降低(P<0.05)。结论干扰circBIRC6表达可通过上调miR-449b-5p表达而抑制胃癌细胞增殖、细胞周期进程及诱导细胞凋亡。Objective To explore the effects of circBIRC6 on the proliferation,cell cycle and apoptosis of gastric cancer cells and its possible mechanism.Methods The specimens of cancer tissues and para-cancerous tissue were collected in 53 patients with gastric cancer treated in Xiaogan Central Hospital Affiliated to Wuhan University of Science and Technology from January 2020 to May 2020.RT-qPCR was used to detect the expression of circBIRC6 and miR-449b-5p in gastric cancer tissues,para-cancerous tissues,human gastric mucosal epithelial cells GES1,and human gastric cancer cells lines(SNU-1,AGS and HS-746T).SNU-1 cells were taken as the research objects and were divided into si-NC group,si-circBIRC6 group,miR-NC group,miR-449b-5p group,si-circBIRC6+anti-miR-NC group and si-circBIRC6+anti-miR-449b-5p group.Proliferation,cell cycle and apoptosis of SNU-1 cells were detected by cell counting kit-8(CCK-8)and flow cytometry assays,respectively.The dual luciferase reporter experiment was conducted to detect the effect of miR-449b-5p overexpression on luciferase activity of wild-type vector WT-circBIRC6.Western blot was used to detect the expression of cleaved caspase-3 protein.Results Compared with para-cancerous tissues,the expression of circBIRC6 in gastric cancer tissues was increased significantly(P<0.05),while the expression of miR-449b-5p was decreased(P<0.05).Compared with GES1 cells,the expressions of circBIRC6 in SNU-1,AGS,and HS-746T cells significantly increased(P<0.05),and the expression of miR-449b-5p decreased(P<0.05).Compared with those in si-NC group,the cell viability and the proportion of S-phase cells were decreased(P<0.05),while the proportion of cells in the G0/G1 phase,the rate of apoptosis and the protein level of cleaved caspase-3 were increased in si-circBIRC6 group(P<0.05).Compared with those in miR-NC group,the cell viability and the S-phase cell ratio decreased(P<0.05),and the G0/G1 phase cell ratio,cell apoptosis rate and the protein level of cleaved caspase-3 significantly increased in miR-449b-5p

关 键 词：胃癌 circBIRC6 miR-449b-5p 细胞增殖 凋亡 细胞周期 

分 类 号：R735.2[医药卫生—肿瘤]