Zhang Guodong, Liu Xiaozhu, Li Yinfeng, Wang Yan, Liang Huankun, Li Kangyan, Li Laiqing, Chen Cuicui, Sun Wenqiao, Ren Shoulei, Zhu Pengfei, Zhang Licheng
Department of Medical Oncology, Cancer Center of Guangzhou Medical University, Guangzhou, Guangdong, China.
School of Light Industry Engineering, Guizhou Institute of Technology, Guiyang, Guizhou, China.
Cancer Sci. 2017 Mar;108(3):408-418. doi: 10.1111/cas.13161.
microRNAs play key roles during various crucial cell processes such as proliferation, migration, invasion and apoptosis. Also, microRNAs have been shown to possess oncogenic and tumor-suppressive functions in human cancers. Here, we describe the regulation and function of miR-149 in colorectal cancer cell lines. miR-149 expression patterns were detected in human colorectal cell lines and tissue samples, and then focused on its role in regulation of cell growth, migration, invasion, and its target gene identification. Furthermore, the function of the target gene of miR-149 was analyzed in vitro and in vivo. miR-149 expression was downregulated in human colorectal cancer HCT116 and SW620 cell lines compared to the normal colon epithelial NCM460 cell line using quantitative real-time polymerase chain reaction methods. Further studies indicated that introduction of miR-149 was able to suppress cell migration and invasion. Then, EphB3 was identified as a direct target gene of miR-149 in colorectal cancer cells. Moreover, experiments in vitro showed that knockdown expression of EphB3 could suppress cell proliferation and invasion, and ectopic expression of EphB3 restored the phenotypes of CRC cell lines transfected with miR149. In addition, silencing of EphB3 significantly affected cycle progression distribution and increased apoptosis in CRC cell lines. Finally, in vivo results demonstrated that knockdown of EphB3 by siRNA inhibited tumor growth. In conclusion,the important role of miR-149 in colorectal cancer progression suggesting that miR-149 may serve as a therapeutic target for colorectal cancer treatment.
微小RNA在各种关键的细胞过程中发挥着关键作用,如增殖、迁移、侵袭和凋亡。此外,微小RNA在人类癌症中已被证明具有致癌和肿瘤抑制功能。在此,我们描述了miR - 149在结肠癌细胞系中的调控及功能。在人类结肠癌细胞系和组织样本中检测miR - 149的表达模式,然后重点研究其在细胞生长、迁移、侵袭调控中的作用及其靶基因鉴定。此外,在体外和体内分析了miR - 149靶基因的功能。使用定量实时聚合酶链反应方法,与正常结肠上皮NCM460细胞系相比,人类结肠癌HCT116和SW620细胞系中miR - 149的表达下调。进一步研究表明,导入miR - 149能够抑制细胞迁移和侵袭。然后,EphB3被鉴定为结肠癌细胞中miR - 149的直接靶基因。此外,体外实验表明,敲低EphB3的表达可抑制细胞增殖和侵袭,EphB3的异位表达恢复了用miR149转染的结肠癌细胞系的表型。此外,沉默EphB3显著影响结肠癌细胞系的细胞周期进程分布并增加凋亡。最后,体内结果表明,通过小干扰RNA敲低EphB3可抑制肿瘤生长。总之,miR - 149在结肠癌进展中起重要作用,提示miR - 149可能作为结肠癌治疗的一个治疗靶点。