Wen Ruhui, Chen Chao, Zhong Xiaohua, Hu Chen
Department of Gastrointestinal Surgery, Huizhou Municipal Central Hospital, Huizhou, Guangdong, China.
Bioengineered. 2021 Dec;12(1):3886-3899. doi: 10.1080/21655979.2021.1940071.
Researchers have demonstrated that long non-coding RNAs (lncRNAs) are vital in colorectal cancer (CRC) progression. Here, we aimed to explore the function of lncRNA PAX6 upstream antisense RNA (PAUPAR) in the development of CRC. In the present study, PAUPAR and microRNA (miR)-17-5p expression levels in CRC tissues and cells were examined using quantitative real-time polymerase chain reaction (qRT-PCR). Western blot analysis was adopted to examine ZNF750 expression at the protein level in CRC cells. CRC cell proliferation was examined by colony formation experiment and 5-Bromo-2-deoxyUridine (BrdU) experiment. CRC cell migration and invasion were assessed by Transwell experiments. Apoptosis was measured using the TUNEL experiment. The targeting relationship between PAUPAR and miR-17-5p was confirmed using dual-luciferase reporter gene and RNA immunoprecipitation (RIP) experiments. We demonstrated that PAUPAR was markedly down-modulated in CRC, and its low expression was significantly related to increased T stage and local lymph node metastasis. Knockdown of PAUPAR enhanced CRC cell proliferation, migration and invasion, and restrained apoptosis relative to controls, whereas PAUPAR overexpression caused the opposite effects. Moreover, rescue experiments showed that miR-17-5p inhibitor could reverse the role of PAUPAR knockdown on the malignant phenotypes of CRC cells. Additionally, PAUPAR could positively regulate the expression of ZNF750 via repressing miR-17-5p. Taken together, these findings suggest that PAUPAR/miR-17-5p/ZNF750 axis is a novel mechanism implicated in CRC progression.
研究人员已证明长链非编码RNA(lncRNA)在结直肠癌(CRC)进展中至关重要。在此,我们旨在探究lncRNA PAX6上游反义RNA(PAUPAR)在CRC发生发展中的作用。在本研究中,采用定量实时聚合酶链反应(qRT-PCR)检测CRC组织和细胞中PAUPAR和微小RNA(miR)-17-5p的表达水平。采用蛋白质印迹分析检测CRC细胞中ZNF750蛋白水平的表达。通过集落形成实验和5-溴-2'-脱氧尿苷(BrdU)实验检测CRC细胞增殖。通过Transwell实验评估CRC细胞迁移和侵袭能力。使用TUNEL实验检测细胞凋亡。通过双荧光素酶报告基因和RNA免疫沉淀(RIP)实验证实PAUPAR与miR-17-5p之间的靶向关系。我们证明PAUPAR在CRC中显著下调,其低表达与T分期增加和局部淋巴结转移显著相关。相对于对照组,敲低PAUPAR可增强CRC细胞增殖、迁移和侵袭能力,并抑制细胞凋亡,而PAUPAR过表达则产生相反作用。此外,挽救实验表明,miR-17-5p抑制剂可逆转PAUPAR敲低对CRC细胞恶性表型的作用。此外,PAUPAR可通过抑制miR-17-5p正向调节ZNF750的表达。综上所述,这些发现表明PAUPAR/miR-17-5p/ZNF750轴是一种参与CRC进展的新机制。