LncRNA GAS5-AS1靶向miR-106a-5p调控结直肠癌细胞增殖、迁移和侵袭  

LncRNA GAS5-AS1 regulates colorectal cancer cell proliferation,migration,and invasion by targeting miR-106a-5p

作　　者：余周 胡满溢 戴剑[1] Zhou Yu;Man-Yi Hu;Jian Dai(Department of Colorectal and Anal Surgery,Jinhua Central Hospital,Jinhua 321000,Zhejiang Province,China)

机构地区：[1]金华市中心医院结直肠肛门外科,浙江省金华市321000

出　　处：《世界华人消化杂志》2024年第7期525-533,共9页World Chinese Journal of Digestology

基　　金：金华市科技计划项目,No.2023-4-068.

摘　　要：背景长链非编码RNA(long non-coding RNA,LncRNA)生长停滞特异性转录本5反义RNA(growth arrest specific transcript 5 antisense RNA,GAS5-AS1)被发现在多种癌症中扮演肿瘤抑制因子的作用.但是其在结直肠癌(colorectal cancer,CRC)进展中的作用和机制尚不清楚.目的探讨LncRNA GAS5-AS1靶向miR-106a-5p在CRC进展中的作用.方法实时荧光定量聚合酶链反应分析LncRNA GAS5-AS1和miR-106a-5p在43对CRC组织和癌旁组织样本中的表达.分别转染pcDNA、pcDNA-LncRNA GAS5-AS1、anti-miR-NC、anti-miR-106a-5p、si-NC、si-LncRNA GAS5-AS1、pcDNA-LncRNA GAS5-AS1+miR-NC、pcDNA-LncRNA GAS5-AS1+miR-106a-5p mimics至HCT8细胞.细胞计数试剂盒-8法检测细胞活力;Transwell小室法检测迁移和侵袭细胞数.通过双荧光素酶报告实验确定LncRNA GAS5-AS1和miR-106a-5p的靶向关系.结果与癌旁组织比较,CRC组织中LncRNA GAS5-AS1表达降低(P<0.05),miR-106a-5p表达升高(P<0.05).与转染pcDNA比较,转染pcDNA-LncRNA GAS5-AS1后HCT8细胞活力、迁移、侵袭、miR-106a-5p表达显著降低(P<0.05).与转染anti-miR-NC比较,转染anti-miR-106a-5p后HCT8细胞活力、迁移和侵袭细胞数显著降低(P<0.05).与转染si-NC比较,转染si-LncRNA GAS5-AS1后miR-106a-5p表达水平显著升高(P<0.05).与转染miR-NC比较,转染miR-106a-5p能显著降低LncRNA GAS5-AS1-WT载体的荧光素酶活性,表明miR-106a-5p是LncRNA GAS5-AS1的直接靶点.与转染pcDNA-LncRNA GAS5-AS1+miR-NC比较,转染pcDNA-LncRNA GAS5-AS1+miR-106a-5p mimics后HCT8细胞活力、迁移和侵袭细胞数显著升高(P<0.05).结论LncRNA GAS5-AS1通过靶向miR-106a-5p来抑制CRC细胞增殖、迁移和侵袭.BACKGROUND The long non-coding RNA(lncRNA)growth arrest specific transcript 5 antisense RNA(GAS5-AS1)has been found to act as a tumor suppressor in a variety of cancers.However,its role and mechanism in colorectal cancer(CRC)progression remain unclear.AIM To investigate the role of the lncRNA GAS5-AS1 targeting miR-106a-5p in the progression of CRC.METHODS GAS5-AS1 and miR-106a-5p expression levels in 43 pairs of CRC tissues and adjacent tissue samples were determined by real-time quantitative PCR.After pcDNA,pcDNA-GAS5-AS1,anti-miR-NC,anti-miR-106a-5p,si-NC,si-GAS5-AS1,pcDNA-GAS5-AS1+miR-NC,and pcDNA-GAS5-AS1+miR-106a-5p mimic were transfected into HCT8 cells,cell counting kit-8 method was used to detect cell viability,and transwell assays were used to detect the number of migrating and invading cells.The target relationship between GAS5-AS1 and miR-106a-5p was determined by dual-luciferase reporter assay.RESULTS Compared with adjacent normal tissues,GAS5-AS1 expression in CRC tissue was decreased(P<0.05),and miR-106a-5p expression was increased(P<0.05).Compared with cells transfected with pcDNA,cell viability,migration,and invasion and miR-106a-5p expression in HCT8 cells were significantly reduced after transfection of with pcDNA-GAS5-AS1(P<0.05).Compared with cells transfected with anti-miR-NC,cell viability,migration,and invasion in HCT8 transfected with anti-miR-106a-5p were significantly reduced(P<0.05).Compared with cells transfected with si-NC,the expression level of miR-106a-5p was significantly increased in cells transfected with si-GAS5-AS1(P<0.05).Compared with cells transfected with miR-NC,transfection of miR-106a-5p significantly reduced the luciferase activity of the GAS5-AS1-WT vector,indicating that miR-106a-5p is the direct target of GAS5-AS1.Compared with cells transfected with pcDNA-GAS5-AS1+miR-NC,cell viability,migration,and invasion were significantly increased in cells transfected with pcDNA-GAS5-AS1+miR-106a-5p mimic(P<0.05).CONCLUSION GAS5-AS1 inhibits CRC cell proliferation,migration,a

关 键 词：结直肠癌 LncRNA GAS5-AS1 miR-106a-5p 细胞增殖 迁移 侵袭 

分 类 号：R735.37[医药卫生—肿瘤]