Wang Wei, Cheng Xia, Zhu Jianhua
Department of Emergency Traumatic Surgery, Shanghai Pudong New District Zhoupu Hospital (Shanghai University of Medicine and Health Sciences Affiliated Zhoupu Hospital), Shanghai 201318, P.R. China.
Graduate School, Dalian Medical University, Dalian, Liaoning 116000, P.R. China.
Oncol Lett. 2021 Mar;21(3):193. doi: 10.3892/ol.2021.12454. Epub 2021 Jan 8.
Colorectal cancer (CRC) is a common digestive system malignancy and a major cause of cancer-associated mortality worldwide. Aberrant expression of long non-coding RNAs has been reported in several types of cancer. The aim of the present study was to investigate the role of ovarian tumor domain containing 6B antisense RNA1 (OTUD6B-AS1) in CRC and its underlying mechanisms. OTUD6B-AS1 expression in CRC cell lines was examined using reverse transcription-quantitative PCR. Furthermore, The Cancer Genome Atlas database was utilized to examine the expression levels of OTUD6B-AS1 in CRC tissues. Following OTUD6B-AS1 overexpression, Cell Counting Kit-8 and colony formation assays were used to detect the proliferation ability of HCT116 cells. The expression levels of proliferation-related protein Ki67 were determined using immunofluorescence staining. Subsequently, Transwell and wound healing assays were used to evaluate the invasion and migration of HCT116 cells, respectively. The expression levels of migration-related proteins (MMP2 and MMP9) were measured using western blotting. Additionally, a luciferase reporter assay was used to verify the potential interaction between OTUD6B-AS1 and microRNA-3171 (miR-3171). Subsequently, rescue assays were performed to clarify the regulatory effects of OTUD6B-AS1 and miR-3171 on CRC development. The results demonstrated that OTUD6B-AS1 expression was low in CRC cells and tissues. Overexpression of OTUD6B-AS1 inhibited the proliferation, invasion and migration of HCT116 cells. Furthermore, miR-3171 was demonstrated to be a direct target of OTUD6B-AS1 using a luciferase reporter assay. The rescue assays revealed that miR-3171 mimics markedly reversed the inhibitory effects of OTUD6B-AS1 overexpression on proliferation, invasion and migration of CRC cells. Overall, these findings demonstrated that OTUD6B-AS1 overexpression inhibited the proliferation, invasion and migration of HCT116 cells via downregulation of miR-3171, suggesting that OTUD6B-AS1 may serve as a novel biomarker for CRC treatment.
结直肠癌（CRC）是一种常见的消化系统恶性肿瘤，也是全球癌症相关死亡的主要原因。在几种类型的癌症中都报道过长链非编码RNA的异常表达。本研究的目的是探讨含卵巢肿瘤结构域6B反义RNA1（OTUD6B-AS1）在结直肠癌中的作用及其潜在机制。使用逆转录定量PCR检测结直肠癌细胞系中OTUD6B-AS1的表达。此外，利用癌症基因组图谱数据库检测结直肠癌组织中OTUD6B-AS1的表达水平。OTUD6B-AS1过表达后，使用细胞计数试剂盒-8和集落形成试验检测HCT116细胞的增殖能力。使用免疫荧光染色测定增殖相关蛋白Ki67的表达水平。随后，分别使用Transwell试验和伤口愈合试验评估HCT116细胞的侵袭和迁移能力。使用蛋白质印迹法测量迁移相关蛋白（MMP2和MMP9）的表达水平。此外，使用荧光素酶报告基因试验验证OTUD6B-AS与微小RNA-3171（miR-3171）之间的潜在相互作用。随后，进行挽救试验以阐明OTUD6B-AS1和miR-3171对结直肠癌发展的调节作用。结果表明，OTUD6B-AS1在结直肠癌细胞和组织中的表达较低。OTUD6B-AS1的过表达抑制了HCT116细胞的增殖、侵袭和迁移。此外，荧光素酶报告基因试验证明miR-3171是OTUD6B-AS1的直接靶点。挽救试验表明，miR-3171模拟物显著逆转了OTUD6B-AS1过表达对结直肠癌细胞增殖、侵袭和迁移的抑制作用。总体而言，这些发现表明OTUD6B-AS1过表达通过下调miR-3171抑制HCT116细胞的增殖、侵袭和迁移，提示OTUD6B-AS1可能作为结直肠癌治疗的新型生物标志物。
