摘要
目的探讨长链非编码RNA(lncRNA)GATA3-反义RNA 1(lncRNA GATA3-AS1)是否通过靶向微小RNA-574-3p(miR-574-3p)调控结直肠癌细胞SW620的增殖、迁移和侵袭。方法采用逆转录-定量聚合酶链反应(RT-qPCR)检测43例结直肠癌组织及其对应癌旁组织中lncRNA GATA3-AS1和miR-574-3p表达情况。根据转染物的不同,将SW620细胞分为si-NC组、si-GATA3-AS1组、miR-NC组、miR-574-3p组、si-GATA3-AS1+anti-miR-NC组、si-GATA3-AS1+anti-miR-574-3p组。采用细胞计数试剂盒8(CCK-8)法检测细胞增殖活性,采用克隆形成实验检测细胞克隆形成数,采用细胞划痕实验检测划痕愈合率,采用Transwell实验检测细胞侵袭能力,采用蛋白质印迹法(Western blot)检测迁移侵袭相关蛋白E-cadherin和N-cadherin表达。采用双荧光素酶报告基因实验检测lncRNA GATA3-AS1与miR-574-3p的靶向关系。结果结直肠癌组织中lncRNA GATA3-AS1表达量高于癌旁组织,miR-574-3p表达量低于癌旁组织,差异有统计学意义(P<0.05)。si-GATA3-AS1组SW620细胞的增殖活性、克隆形成数、划痕愈合率、侵袭细胞数和N-cadherin蛋白表达量均低于si-NC组,E-cadherin蛋白表达量高于si-NC组,差异有统计学意义(P<0.05)。miR-574-3p组SW620细胞的增殖活性、克隆形成数、划痕愈合率、侵袭细胞数和N-cadherin蛋白表达量均低于miR-NC组,E-cadherin蛋白表达量高于miR-NC组,差异有统计学意义(P<0.05)。lncRNA GATA3-AS1靶向调控miR-574-3p的表达。si-GATA3-AS1+anti-miR-574-3p组SW620细胞的增殖活性、克隆形成数、划痕愈合率、侵袭细胞数和N-cadherin蛋白表达量均高于si-GATA3-AS1+anti-miR-NC组,E-cadherin蛋白表达量低于si-GATA3-AS1+anti-miR-NC组,差异有统计学意义(P<0.05)。结论lncRNA GATA3-AS1在结直肠癌中表达上调,可通过靶向调控miR-574-3p促进结直肠癌细胞SW620的增殖、迁移和侵袭。
Objective To explore whether long noncoding RNA(lncRNA)GATA3-antisense RNA 1(lncRNA GATA3-AS1)regulates the proliferation,migration,and invasion of colorectal cancer SW620 cells by targeting micror NA-574-3p(miR-574-3p).Methods Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was used to determine the expressions of lncRNA GATA3-AS1 and miR-574-3p in colorectal cancer tissues of 43 cases and para-cancerous tissue.The SW620 cells were divided into si-NC group,si-GATA3-AS1 group,miR-NC group,miR-574-3p group,si-GATA3-AS1+anti-miR-NC group,si-GATA3-AS1+anti-miR-574-3p group according to different transfection.The cell counting kit-8(CCK-8)method was used to detect cell proliferation activity,the clone formation experiment to measure the number of cell clones,the scratch test to examine the scratch healing rate,the Transwell experiment to analyze cell invasion,and the Western blot to determine the expression of migration and invasion related proteins E-cadherin and N-cadherin.The dual luciferase reporter experiment was used to detect the targeting relationship between lncRNA GATA3-AS1 and miR-574-3p.Results The expression of lncRNA GATA3-AS1 in colorectal cancer tissues was higher,and the expression of miR-574-3p decreased compared with that in the adjacent tissues(P<0.05).SW620 cells in the si-GATA3-AS1 group had lower proliferation activity,the number of clone formation,scratch healing rate,the number of invasive cell,and N-cadherin protein expression than the si-NC group,but the expression of E-cadherin protein was higher than that of si-NC group(P<0.05).SW620 cell proliferation activity,the number of clone formation,scratch healing rate,the number of invasive cells,and N-cadherin protein expression in the miR-574-3p group were lower than those in the miR-NC group,and the E-cadherin protein expression level was higher than that of the miR-NC group(P<0.05).LncRNA GATA3-AS1 targeted and regulated the expression of miR-574-3p.The proliferation activity,the number of clone cells,scratch healing rat
作者
盛华明
李森
邓立春
姚勇
SHENG Huaming;LI Sen;DENG Lichun;YAO Yong(Department of Oncology,Jiangyin Hospital Affiliated to School of Medicine of Southeast University,Nanjing,Jiangsu,214400;Department of Anorectal Surgery,Jiangyin Hospital Affiliated to School of Medicine of Southeast University,Nanjing,Jiangsu,214400)
出处
《实用临床医药杂志》
2023年第14期51-57,共7页
Journal of Clinical Medicine in Practice
基金
国家中医药管理局名医验方评价与转化重点实验室项目(NZYJDMF-2018001)
北京希思科临床肿瘤学研究基金会(Y-2019Genecast-006)
江苏省无锡市卫生健康委科研项目(MS201922)。
