长链非编码RNA OPA相互作用蛋白5反向转录序列1靶向调控微RNA-128-3p对结直肠癌细胞增殖、侵袭和转移的影响  

The molecular mechanism of long non-coding RNA OIP5-AS1 targeting miR-128-3p to regulate the proliferation,invasion and metastasis of colorectal cancer cells

作　　者：郑留昌 崔发财 郑培明 赵伟锋 ZHENG Liuchang;CUI Facai;ZHENG Peiming;ZHAO Weifeng(Clinical Laboratory,The People's Hospital of Zhengzhou University,Zhengzhou,Henan 450003,China;Department of Medical Oncology,The People's Hospital of Zhengzhou University,Zhengzhou,Henan 450003,China)

机构地区：[1]郑州大学人民医院检验科,河南郑州450003 [2]郑州大学人民医院肿瘤内科,河南郑州450003

出　　处：《安徽医药》2024年第2期259-265,共7页Anhui Medical and Pharmaceutical Journal

基　　金：国家自然科学基金项目(81802094)。

摘　　要：目的探究长链非编码RNA OPA相互作用蛋白5反向转录序列1(lncRNA OIP5-AS1)在结直肠癌中的表达情况及靶向调控微RNA-128-3p(miR-128-3p)对结直肠癌细胞增殖、侵袭和转移的影响。方法采用实时荧光定量PCR(qPCR)定量分析2017年1月至2018年12月在郑州大学人民医院住院并进行手术治疗的38例结直肠癌病人癌组织及相应癌旁组织、结直肠癌细胞系(SW480、SW620、HT-29和LoVo)及人正常结直肠黏膜细胞FHC中lncRNA OIP5-AS1和miR-128-3p的表达。将SW620细胞设为si-OIP5-AS1组、si-NC组、miR-128-3p mimic组、mimic-NC组、miR-128-3p inhibitor+si-OIP5-AS1组和inhibitor-NC+siOIP5-AS1组,采用细胞计数试剂盒(CCK-8)实验和克隆形成实验检测细胞增殖能力,采用划痕实验和transwell实验检测细胞侵袭与迁移能力,采用蛋白质印迹法检测E2F1、细胞周期蛋白D1(Cyclin D1)、波形蛋白(Vimentin)、N-钙黏蛋白(N-cadherin)和E-钙黏蛋白(E-cadherin)蛋白表达,采用双萤光素酶报告实验验证lncRNA OIP5-AS1与miR-128-3p的靶向关系。结果与癌旁组织相比较,lncRNA OIP5-AS1在结直肠癌组织中表达显著升高(1.42±0.40比0.98±0.29,P<0.05),miR-128-3p表达显著降低(0.72±0.21比1.52±0.39,P<0.05),且两者在结直肠癌组织中的表达呈负相关;与FHC细胞(1.05±0.10)相比较,lncRNA OIP5-AS1在结直肠癌细胞株[SW480(1.60±0.25)、SW620(2.19±0.33)、HT-29(1.62±0.23)和LoVo(1.95±0.20)]中的表达显著增高(均P<0.05),miR-128-3p表达(0.70±0.15、0.46±0.03、0.59±0.14、0.74±0.09比1.02±0.11)显著降低(均P<0.05)。lncRNA OIP5-AS1与miR-128-3p为相互作用靶基因,lncRNA OIP5-AS1表达下调或miR-128-3p表达上调均能降低vimentin、N-cadherin、E2F1和cyclin D1蛋白表达,增高E-cadherin蛋白表达,抑制SW620细胞的增殖、侵袭和迁移。在转染si-OIP5-AS1的同时转染miR-128-3p inhibitor可增加vimentin、N-cadherin、E2F1和cyclin D1蛋白表达,降低E-cadherin蛋白的表达,逆转lncRNA OIP5-AS1表达下Objective To investigate the expression of long non-coding RNA(lncRNA)Opa interacting protein 5 antisense RNA l(OIP5-AS1)in colorectal cancer(CRC)and its effect on the proliferation,invasion and metastasis of CRC cells by targeting miR-128-3p.Methods The expressions of lncRNA OIP5-AS1 and miR-128-3p in cancer tissues and adjacent normal tissues of patients admitted to The People's Hospital of Zhengzhou University and undergoing surgery from January 2017 to December 2018,CRC cell lines(SW480,SW620,HT-29 and LoVo)and human normal colorectal mucosal cells FHC were quantitatively analyzed by qPCR.SW620 cells were transfected with si-OIP5-AS1,si-NC,miR-128-3p mimics,mimics-NC,miR-128-3p inhibitor+si-OIP5-AS1 or inhibitor-NC+si-OIP5-AS1,the proliferation,invasion and migration of SW620 cells were detected by clone formation assay,CCK-8 assay,woundhealing assay and transwell assay,respectively.The expressions of E2F1,cyclin D1,Vimentin,N-cadherin and E-cadherin were detect⁃ed by Western blotting assay.Dual luciferase reporter assay was used to verify the targeting relationship between lncRNA OIP5-AS1 and miR-128-3p.Results Compared with adjacent normal tissues,the expression of lncRNA OIP5-AS1 in CRC tissues was signifi⁃cantly increased(1.42±0.40 vs.0.98±0.29,P<0.05),and the expression of miR-128-3p was significantly decreased(0.72±0.21 vs.1.52±0.39,P<0.05),and the expressions of lncRNA OIP5-AS1 and miR-128-3p in CRC tissues were negatively correlated.Compared with FHC cells,the expressions of lncRNA OIP5-AS1 in CRC cell lines[(1.05±0.10)vs.SW480(1.60±0.25),SW620(2.19±0.33),HT-29(1.62±0.23)and LoVo(1.95±0.20)]were significantly increased,and the expressions of miR-128-3p were significantly decreased[(0.70±0.15),(0.46±0.03),(0.59±0.14),(0.74±0.09)vs.(1.02±0.11)].LncRNA OIP5-AS1 and miR-128-3p were target genes interacting with each other.Down-regulation of lncRNA OIP5-AS1 or up-regulation of miR-128-3p could decrease the expressions of Vimentin,N-cadherin,E2F1 and cyclin D1,increase the expression of E-

关 键 词：结直肠肿瘤 侵袭 转移 微RNA-128-3p 长链非编码RNA Opa相互作用蛋白5-反向RNA1 

分 类 号：R735.34[医药卫生—肿瘤]