lncRNA ZSWIM8-AS1对结直肠癌细胞RKO增殖、凋亡、迁移和放射敏感性的影响  

Effects of lncRNA ZSWIM8-AS1 on proliferation,apoptosis,migration and radiosensitivity of colorectal cancer RKO cells

作　　者：侯歌[1] 张家杰 肖陈虎 宋锐[1] 黄洋洋 袁金金[1] 柴婷[1] 刘宗文[1] HOU Ge;ZHANG Jiajie;XIAO Chenhu;SONG Rui;HUANG Yangyang;YUAN Jinjin;CHAI Ting;LIU Zongwen(Department of Radiation Oncology,the Second Affiliated Hospital,Zhengzhou University,Zhengzhou 450014;College of Life Science,Northwest University,Xian 710069)

机构地区：[1]郑州大学第二附属医院肿瘤放疗科,郑州450014 [2]西北大学生命科学学院,西安710069

出　　处：《郑州大学学报（医学版）》2021年第4期485-492,共8页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省医学科技攻关计划联合共建项目(2018020161)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)ZSWIM8-AS1对结直肠癌细胞增殖、凋亡、迁移和放射敏感性的影响及作用机制。方法:选取25例结直肠癌组织和对应的癌旁组织,采用qRT-PCR法检测组织中ZSWIM8-AS1和miR-15b表达水平。体外培养结直肠癌细胞RKO,双荧光素酶报告基因实验验证ZSWIM8-AS1和miR-15b调控关系。分组培养RKO细胞(pcDNA组、pcDNA-ZSWIM8-AS1组、anti-miR-NC组、anti-miR-15b组、pcDNA-ZSWIM8-AS1+miR-NC组和pcDNA-ZSWIM8-AS1+miR-15b组),采用细胞计数试剂盒-8(CCK-8)法检测各组细胞存活率,双染法检测各组细胞凋亡率,Transwell小室检测各组细胞迁移,克隆形成实验检测各组细胞放射敏感性,Western blot法检测各组细胞中Ki-67、Cleaved Caspase-3、N-cadherin、E-cadherin的蛋白表达水平。结果:与癌旁组织比较,结直肠癌组织中ZSWIM8-AS1表达降低(P<0.001),miR-15b表达升高(P<0.001)。ZSWIM8-AS1在RKO细胞中负调控miR-15b表达。与pcDNA组或anti-miR-NC组比较,pcDNA-ZSWIM8-AS1组和anti-miR-15b组细胞存活率、迁移数及Ki-67和N-cadherin蛋白表达均降低(P均<0.001),凋亡率及Cleaved Caspase-3和E-cadherin蛋白表达升高(P均<0.001),增敏比分别为1.747、1.977。与pcDNA-ZSWIM8-AS1+miR-NC组比较,pcDNA-ZSWIM8-AS1+miR-15b组细胞存活率、迁移数及Ki-67和N-cadherin蛋白表达升高(P均<0.001),凋亡率及Cleaved Caspase-3和E-cadherin蛋白表达降低(P均<0.001),增敏比为0.645。结论:ZSWIM8-AS1在结直肠癌组织中表达降低,上调其表达可通过靶向负调控miR-15b抑制结直肠癌细胞RKO增殖和迁移,并促进细胞凋亡和对放射线的敏感性。Aim:To investigate the effects and mechanism of lncRNA ZSWIM8-AS1 on proliferation,apoptosis,migration and radiosensitivity of colorectal cancer cells.Methods:Twenty-five cases of colorectal cancer tissue and corresponding paracancerous tissue were selected,and qRT-PCR was used to detecte the expression levels of ZSWIM8-AS1 and miR-15b in the samples.Colorectal cancer RKO cells were cultured in vitro,and the dual luciferase reporter gene experiment was used to verify the regulatory relationship between ZSWIM8-AS1 and miR-15b.RKO cells were allocted into pcDNA group,pcDNA-ZSWIM8-AS1 group,anti-miR-NC group,anti-miR-15b group,pcDNA-ZSWIM8-AS1+miR-NC group and pcDNA-ZSWIM8-AS1+miR-15b group,then CCK-8 method was used to detecte cell survival rate,Annexin V-FITC/PI double staining to detecte cell apoptosis rate,Transwell chamber to detecte cell migration,clone formation experiment to detecte cell radiosensitivity,and Western blot to detecte the protein expression levels of Ki-67,Cleaved Caspase-3,N-cadherin,and E-cadherin.Results:Compared with adjacent tissue,the expression of ZSWIM8-AS1 in colorectal cancer tissue decreased(P<0.001),and the expression of miR-15b increased(P<0.001).ZSWIM8-AS1 negatively regulated miR-15b expression in RKO cells.Compared with the pcDNA group or anti-miR-NC group,the cell survival rate,number of migration cells,and the protein expression levels of Ki-67 and N-cadherin in the pcDNA-ZSWIM8-AS1 group and anti-miR-15b group were decreased(all P<0.001),while the apoptosis rate and the protein expression levels of Cleaved Caspase-3 and E-cadherin were increased(all P<0.001),and the sensitization ratio was 1.747,1.977,respectively.Compared with the pcDNA-ZSWIM8-AS1+miR-NC group,the cell survival rate,number of migration cells,and the protein expression levels of Ki-67 and N-cadherin in the pcDNA-ZSWIM8-AS1+miR-15b group were increased(all P<0.001),while the apoptosis rate and the protein expression levels of Cleaved Caspase-3 and E-cadherin were decreased(all P<0.001),and the sensitization ra

关 键 词：结直肠癌 ZSWIM8-AS1 miR-15b 放射敏感性 RKO细胞 

分 类 号：R735.3[医药卫生—肿瘤]