Chen Rui, Zhou Shenkang, Chen Jianhui, Lin Senbin, Ye Feifei, Jiang Pinlu
Department of Gastroenterology, Taizhou Hospital of Zhejiang Province, Taizhou 317000, People's Republic of China.
Department of Emergency, Taizhou Hospital of Zhejiang Province, Taizhou 317000, People's Republic of China.
Cancer Manag Res. 2020 Dec 22;12:13137-13148. doi: 10.2147/CMAR.S274447. eCollection 2020.
Colorectal cancer (CRC) is a common severe disease around the world. The merging papers reported that long noncoding RNAs (lncRNAs) took part in the diversified pathological processes of CRC. This study aimed to uncover the role and the potential mechanism of lncRNA bladder cancer-associated transcript 1 (BLACAT1) in CRC progression.
LncRNA BLACAT1, micro-519d-3p (miR-519d-3p), and cAMP-responsive element binding protein 1 () levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR) in CRC tissues and cells. The bio-functional effects were examined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), flow cytometry assay, and transwell assay. The susceptibility testing was determined by oxaliplatin (OXA) administration. The potential binding sites between miR-519d-3p and BLACAT1 or were predicted by online software starBase and confirmed by dual-luciferase reporter analysis. The relative proteins expression in CRC cells was determined by Western blot analysis. Xenograft tumor model was used to evaluate biological function of BLACAT1 in vivo.
The expression of BLACAT1 was promoted in CRC tissues and cells, and correlated to the TNM (tumor, node, metastasis) stage, distant metastasis, and overall survival rate. Silencing of BLACAT1 limited the proliferation, migration, and invasion, facilitated the apoptosis, and re-sensitized OXA-resistance in CRC cells. MiR-519d-3p was a target of BLACAT1. Furthermore, miR-519d-3p deletion reversed the positive effects of BLACAT1 deletion on CRC cells. Moreover, our data showed that miR-519d-3p directly targeted and BLACAT1 sponged miR-519d-3p to regulate expression. Besides, disrupted the bio-functional results above from BLACAT1 suppression. Additionally, BLACAT1 knockdown promoted CRC cells sensitivity to OXA in vivo.
BLACAT1 mediated the progression of CRC and OXA-resistance by miR-519d-3p/ axis.
结直肠癌（CRC）是全球常见的严重疾病。合并的文献报道长链非编码RNA（lncRNAs）参与了CRC的多种病理过程。本研究旨在揭示lncRNA膀胱癌相关转录本1（BLACAT1）在CRC进展中的作用及潜在机制。
采用定量实时聚合酶链反应（qRT-PCR）检测CRC组织和细胞中lncRNA BLACAT1、微小RNA-519d-3p（miR-519d-3p）和环磷酸腺苷反应元件结合蛋白1（）的水平。通过3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐（MTT）、流式细胞术检测和Transwell检测来研究其生物学功能。通过给予奥沙利铂（OXA）进行药敏试验。利用在线软件starBase预测miR-519d-3p与BLACAT1或之间的潜在结合位点，并通过双荧光素酶报告基因分析进行验证。采用蛋白质印迹分析确定CRC细胞中相关蛋白的表达。利用异种移植瘤模型在体内评估BLACAT1的生物学功能。
BLACAT1在CRC组织和细胞中表达上调，且与TNM（肿瘤、淋巴结、转移）分期、远处转移及总生存率相关。沉默BLACAT1可限制CRC细胞的增殖、迁移和侵袭，促进细胞凋亡，并使CRC细胞对OXA重新敏感。MiR-519d-3p是BLACAT1的靶标。此外，miR-519d-3p缺失可逆转BLACAT1缺失对CRC细胞的积极作用。而且，我们的数据表明miR-519d-3p直接靶向，而BLACAT1通过海绵吸附miR-519d-3p来调节的表达。此外，破坏了上述BLACAT1抑制所产生的生物学功能结果。另外，BLACAT1基因敲低可提高CRC细胞在体内对OXA的敏感性。
BLACAT1通过miR-519d-3p/轴介导CRC的进展及对OXA的耐药性。
