摘要
目的探讨lncRNA LINC00909是否通过靶向miR-548-3p而影响结直肠癌细胞放射敏感性。方法采用qRT-PCR检测结直肠癌组织、癌旁组织中LINC00909、miR-584-3p的表达量;体外培养结直肠癌细胞SW480、SW620,分别将si-NC、si-LINC00909、miR-NC、miR-584-3p mimics、si-LINC00909与anti-miR-NC、si-LINC00909与anti-miR-584-3p转染至SW480、SW620细胞,用4 Gy照射细胞;克隆形成实验检测细胞存活分数及放射增敏比;MTT检测细胞增殖;Transwell小室实验检测细胞迁移及侵袭;双荧光素酶报告实验验证LINC00909、miR-584-3p的靶向关系。裸鼠皮下移植瘤实验检测干扰LINC00909表达或抑制miR-584-3p表达对照射后移植瘤重量的影响。结果结直肠癌组织中LINC00909的表达水平显著升高(P<0.05),miR-584-3p的表达水平显著降低(P<0.05);干扰LINC00909表达或miR-584-3p过表达后细胞存活分数明显降低(P<0.05),放射增敏比分别为2.017、1.762,并可抑制增殖、迁移及侵袭(P<0.05);双荧光素酶报告实验证实LINC00909可靶向结合miR-584-3p;干扰LINC00909表达后移植瘤重量显著降低(P<0.05)。共转染anti-miR-584-3p后移植瘤重量显著升高(P<0.05)。结论干扰LINC00909表达可通过上调miR-548-3p的表达而减弱结直肠癌细胞增殖、迁移及侵袭能力从而增强细胞放射敏感性。
Objective To investigate whether lncRNA LINC00909 affected the radiosensitivity of colorectal cancer cells by targeting miR-548-3p.Methods The expression levels of LINC00909 and miR-584-3p in colorectal cancer and adjacent tissues were detected by qRT-PCR.The colorectal cancer cells SW480 and SW620 were cultured in vitro,and transfected with si-NC,si-LINC00909,miR-NC,miR-584-3p mimics,si-LINC00909,and anti-miR-NC and si-LINC00909,and anti-miR-584-3p,respectively.The cells were irradiated with a dose of 4 Gy.The cell survival fraction and sensitization enhancement ratio(SER)were detected by clone formation assay.Cell proliferation was detected by MTT assay.Cell migration and invasion were assessed by Trans well chamber assay.The targeting relationship between LINC00909 and miR-584-3p was confirmed by dual luciferase reporter assay.The effect of interfering with the expression of LINC00909 or inhibiting the expression of miR-584-3p on the weight of the xenograft tumor after irradiation was evaluated by subcutaneous xenograft experiment in nude mice.Results The expression level of LINC00909 in colorectal cancer tissues was significantly up-regulated(P<0.05),whereas the expression level of miR-584-3p was significantly down-regulated(P<0.05).After interfering with the expression of LINC00909 or miR-584-3p overexpression,the cell survival fraction score was significantly reduced(P<0.05),the SERs were 2.017 and 1.762,and cell proliferation,migration and invasion were suppressed(all P<0.05).Dual luciferase reporter assay confirmed that LINC00909 could target and bind to miR-584-3p.After interfering with the expression of LINC00909,the weight of the transplanted tumor was significantly reduced(P<0.05),whereas the weight of the transplanted tumor was significantly increased after co-transfection with anti-miR-584-3p(P<0.05).Conclusion Interfering with the expression of LINC00909 can inhibit the proliferation,migration and invasion ability of colorectal cancer cells by up-regulating the expression of miR-548-3p,thereby enha
作者
侯歌
肖陈虎
陈晓娟
黄洋洋
王成
柴婷
宋锐
袁金金
刘宗文
Hou Ge;Xiao Chenhu;Chen Xiaojuan;Huang Yangyang;Wang Cheng;Chai Ting;Song Rui;Yuan Jinjin;Liu Zongwen(Department of Radiation Oncology,The Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)
出处
《中华放射肿瘤学杂志》
CSCD
北大核心
2020年第12期1110-1117,共8页
Chinese Journal of Radiation Oncology
基金
2018年度河南省医学科技攻关计划联合共建项目(2018020161)。
