Wu Xiongjian, Zhu Haiyan, Xie Yuan, Gu Xiaoxiang, Zhang Lei, Huang Lixing
Department of Gastroenterology, the First Affiliated Hospital of Gannan Medical University, Ganzhou, China.
J BUON. 2020 Jan-Feb;25(1):194-201.
To uncover the potential function of long non-coding RNA (lncRNA) ZEB2-AS1 in the progression of colorectal cancer (CRC), and its underlying mechanism.
Relative level of ZEB2-AS1 in CRC tissues and matched normal ones was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Correlation between ZEB2-AS1 level and survival of CRC patients was analyzed by Kaplan-Meier method. Regulatory effects of ZEB2-AS1 on cellular behaviors of CRC cells were evaluated. The interactions between ZEB2-AS1 with LSD1 and EZH2 were explored by RNA immunoprecipitation (RIP) assay. 5-Ethynyl-2'- deoxyuridine (EdU) assay was performed to elucidate the roles of ZEB2-AS1, LSD1 and EZH2 on the proliferative ability of CRC cells. Finally, Spearman's correlation analysis was performed to analyze the relationship between ZEB2-AS1 level and expressions of proliferation- and invasion-related genes.
ZEB2-AS1 was upregulated in CRC tissues relative to matched controls. Its level remained higher in CRC patients with ≥6 cm in tumor size, nodal metastasis and stage III-IV. CRC patients with low-level ZEB2-AS1 presented worse survival compared with those with high-level ZEB2-AS1. QRT-PCR data showed higher abundance of ZEB2-AS1 in CRC cell lines than colonic epithelial cell line. Knockdown of ZEB2-AS1 attenuated the proliferative, migratory and invasive abilities, but induced apoptosis of DLD1 and SW620 cells. RIP assay demonstrated the interaction between ZEB2-AS1 and LSD1, EZH2. Moreover, EdU assay revealed that transfection of sh-ZEB2-AS1 attenuated the proliferative ability, which was further reduced after co-transfection of sh-LSD1 or sh-EZH2. Finally, correlation analysis showed that mRNA level of ZEB2-AS1 was positively correlated to those of LSD1, EZH2, MMP9, MMP12 and KRAS, but negatively correlated to KLF2.
LncRNA ZEB2-AS1 is upregulated in CRC. It accelerates CRC cells to proliferate via interacting with EZH2 and LSD1, thus promoting the progression of CRC.
揭示长链非编码RNA（lncRNA）ZEB2-AS1在结直肠癌（CRC）进展中的潜在功能及其潜在机制。
采用定量实时聚合酶链反应（qRT-PCR）检测CRC组织及配对正常组织中ZEB2-AS1的相对水平。采用Kaplan-Meier法分析ZEB2-AS1水平与CRC患者生存率的相关性。评估ZEB2-AS1对CRC细胞生物学行为的调控作用。通过RNA免疫沉淀（RIP）实验探究ZEB2-AS1与赖氨酸特异性去甲基化酶1（LSD1）和增强子zeste同源物2（EZH2）之间的相互作用。采用5-乙炔基-2'-脱氧尿苷（EdU）实验阐明ZEB2-AS1、LSD1和EZH2对CRC细胞增殖能力的作用。最后，进行Spearman相关性分析，以分析ZEB2-AS1水平与增殖和侵袭相关基因表达之间的关系。
与配对对照组相比，CRC组织中ZEB2-AS1上调。在肿瘤大小≥6 cm、有淋巴结转移及III-IV期的CRC患者中，其水平仍然较高。ZEB2-AS1低水平的CRC患者比ZEB2-AS1高水平的患者生存率更差。qRT-PCR数据显示，CRC细胞系中ZEB2-AS1的丰度高于结肠上皮细胞系。敲低ZEB2-AS1可减弱DLD1和SW620细胞的增殖、迁移和侵袭能力，但诱导其凋亡。RIP实验证明了ZEB2-AS1与LSD1、EZH2之间的相互作用。此外，EdU实验表明，转染sh-ZEB2-AS1可减弱增殖能力，在共转染sh-LSD1或sh-EZH2后增殖能力进一步降低。最后，相关性分析表明，ZEB2-AS1的mRNA水平与LSD1、EZH2、基质金属蛋白酶9（MMP9）、基质金属蛋白酶12（MMP12）和KRAS呈正相关，但与 Kruppel样因子2（KLF2）呈负相关。
lncRNA ZEB2-AS1在CRC中上调。它通过与EZH2和LSD1相互作用，加速CRC细胞增殖，从而促进CRC进展。
