摘要
目的探讨长链非编码RNA（lincRNA）－ROR作为一个内源竞争RNA（ceRNA）结合miR－145对下游干性相关基因Oct4、Sox2和Nanog的表达，及对结直肠癌干细胞生物学特性的影响，并阐明这种分子调控网络的临床意义。方法收集2014年至2016年间河南省南阳市中心医院及南阳市第二人民医院胃肠外科结直肠癌患者的癌组织及癌旁组织共52例，采用即时荧光定量聚合酶链反应（qPCR）检测52例临床患者结直肠癌组织标本及结直肠癌细胞系中lincRNA－ROR、miR－145的表达；分析lincRNA－ROR和miR－145的表达与结直肠癌临床病理特征的相关性；流式细胞术从SW1116中分离CD44－CD133－和CD44＋CD133＋细胞；qPCR检测细胞中CD44、CD133、Oct4、Sox2、Nanog表达及CD44＋CD133＋细胞贴壁黏附后CD44、CD133、lincRNA－ROR、miR－145表达；生物信息学分析miR－145与lincRNA－ROR及核心转录因子Oct4、Sox2、Nanog之间靶向调控关系，双荧光素酶报告基因实验、qPCR、Western blot进行验证；噻唑蓝法、克隆形成检测沉默lincRNA－ROR对结直肠癌干细胞增殖及化疗敏感性的影响。结果结直肠癌组织中lincRNA－ROR高表达，miR－145低表达，两者呈显著负相关（P〈0.05）；lincRNA－ROR表达与肿瘤大小、淋巴结转移、远端转移有关（P〈0.05），miR－145表达与肿瘤大小、肿瘤位置有关（P〈0.05）；流式细胞术成功分选出CD44＋CD133＋和CD44－ CD133－细胞，CD44＋CD133＋细胞中CD44、CD133、Oct4、Sox2、Nanog、lincRNA－ROR较CD44－ CD133－细胞高表达，miR－145低表达（P〈0.05）；贴壁黏附后CD44、CD133、lincRNA－ROR表达明显降低，miR－145表达增加（P〈0.05）；lincRNA－ROR能够结合miR－145调控Oct4、Sox2和Nanog表达；沉默lincRNA－ROR可显著抑制结直肠癌干细胞增殖克隆形成能力，增加其对顺铂、紫杉醇敏感性。结论lincRNA－ROR结合miR－145调控核心转录因子Oct4、Sox2�
ObjectiveTo investigate the impact of lincRNA-ROR, a ceRNA by binding miR-145 on the expression of the downstream genes Oct4, Sox2 and Nanog, and related biological characteristics of colon cancer stem cells, and to elucidate the clinical significance of this molecular regulatory network.MethodsFifty-two cases of colorectal cancer tissue and adjacent tissue were collected at Nanyang City Central Hospital and Nanyang Second Hospital, Henan Province, from 2014 to 2016. Real-time quantitative polymerase chain reaction （qPCR） was used to detect the expression of lincRNA-ROR and miR-145 in colorectal cancer tissue and isolated colon cancer cells. The correlation between the expression of lincRNA-ROR, miR-145 and the clinicopathologic features of colon cancer was performed. CD44-CD133- and CD44＋ CD133＋ cells were isolated from SW1116 by using flow cytometry. The expression of CD44, CD133, Oct4, Sox2, Nanog, lincRNA-ROR and miR-145 in cells were detected by qPCR. The relationship between lincRNA-ROR, miR-145, Oct4, Sox2 and Nanog was analyzed by bioinformatics, dual luciferase reporter assay, qPCR and Western blot. The effects of silencing lincRNA-ROR on the proliferation and chemosensitivity of colon cancer stem cells were detected by MTT, colony formation.ResultsLincRNA-ROR was frequently up-regulated and inversely correlated with miR-145 down-regulation in the colon cancer specimens（P〈0.05）. LincRNA-ROR was related to tumor size, lymph node involvement and distant metastasis（P〈0.05）, and miR-145 was found related to tumor size and tumor location（P〈0.05）. CD44＋ CD133＋ cells were successfully isolated from SW1116 by flow cytometry. The levels of CD44, CD133, Oct4, Sox2, Nanog, lincRNA-ROR in CD44＋ CD133＋ cells were significantly increased, while miR-145 was decreased compared with CD44-CD133-cells（P〈0.05）. The levels of CD44, CD133, lnc-ROR in CD44＋ CD133＋ cells were significantly reduced upon cell adherence, while miR-145 was significantly increased（P〈0.05）. Bioinforma
作者
闫振宇
孙新超
Yan Zhenyu , Sun Xinchao(Department of Pathology, Nanyang City Center Hospital, Henan Province, Nanyang 473009, Chin)
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2018年第4期284-290,共7页
Chinese Journal of Pathology
基金
河南省医学科技攻关项目（201602377）
