Liu Xing, Zhang Yexiang, Wang Yan, Bian Chao, Wang Fengji
Department of Anorectal Surgery, Jining NO. 1 People's Hospital, Jining, 272000 Shandong China.
Department of Surgery, Second People's Hospital, Rencheng District, Jining, 272061 Shandong China.
Cancer Cell Int. 2020 Jul 24;20:340. doi: 10.1186/s12935-020-01425-2. eCollection 2020.
Long non-coding RNAs (lncRNAs) have been certified to be involved in the occurrence and growth of diverse cancers, including CRC. The purpose of the research was to explore the effects of lncRNA KCNQ1 overlapping transcript 1 (KCNQ1OT1) on proliferation, migration, invasion, and apoptosis in CRC cells and its mechanism.
The levels of KCNQ1OT1 and miR-329-3p were examined by quantitative real-time polymerase chain reaction (qRT-PCR) in CRC tissues and cells. The mRNA and protein levels of catenin delta-1 (CTNND1) were measured by qRT-PCR and western blot analysis, respectively. The targets of KCNQ1OT1 and miR-329-3p were predicted by online software and confirmed by luciferase reporter assay. The cell proliferation, migration, invasion, and apoptosis were examined using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), transwell, and apoptosis assay. The expression levels of CyclinD1, Bcl-2, MMP9, Cleaved-casp-3, and E-cadherin in SW480 and LS1034 cells were gauged by western blot analysis. Xenograft tumor model was structured to prove the biological role of KCNQ1OT1 of CRC in vivo.
The levels of KCNQ1OT1 and CTNND1 were significantly increased in CRC tissues and cells. Knockdown of KCNQ1OT1 suppressed proliferation, migration, invasion, and induced apoptosis in CRC cells. Conversely, CTNND1 overexpression reversed the impact of KCNQ1OT1 knockdown on CRC cells. Moreover, CTNND1 was verified as a direct target of miR-329-3p, and miR-329-3p could specially bind to KCNQ1OT1. Also, the down-regulation of KCNQ1OT1 triggered the CRC progress by up-regulating CTNND1 expression in CRC cells. Besides, KCNQ1OT1 knockdown inhibited CRC tumor growth through the miR-329-3p/CTNND1 axis in vivo.
Our results indicated that KCNQ1OT1 could positively regulate CTNND1 expression by sponging miR-329-3p, thereby boosting the progression of CRC. Our findings provided the underlying therapy targets for CRC.
长链非编码RNA（lncRNA）已被证实参与多种癌症的发生和发展，包括结直肠癌（CRC）。本研究旨在探讨lncRNA KCNQ1重叠转录本1（KCNQ1OT1）对CRC细胞增殖、迁移、侵袭和凋亡的影响及其机制。
采用定量实时聚合酶链反应（qRT-PCR）检测CRC组织和细胞中KCNQ1OT1和miR-329-3p的水平。分别通过qRT-PCR和蛋白质免疫印迹分析检测连环蛋白δ-1（CTNND1）的mRNA和蛋白质水平。通过在线软件预测KCNQ1OT1和miR-329-3p的靶标，并通过荧光素酶报告基因检测进行验证。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）、Transwell和凋亡检测法检测细胞增殖、迁移、侵袭和凋亡情况。通过蛋白质免疫印迹分析检测SW480和LS1034细胞中细胞周期蛋白D1（CyclinD1）、Bcl-2、基质金属蛋白酶9（MMP9）、裂解的半胱天冬酶-3（Cleaved-casp-3）和E-钙黏蛋白的表达水平。构建异种移植肿瘤模型以证明KCNQ1OT1在体内对CRC的生物学作用。
CRC组织和细胞中KCNQ1OT1和CTNND1水平显著升高。敲低KCNQ1OT1可抑制CRC细胞的增殖、迁移、侵袭并诱导其凋亡。相反，CTNND1过表达可逆转KCNQ1OT1敲低对CRC细胞的影响。此外，CTNND1被证实为miR-329-3p的直接靶标，且miR-329-3p可特异性结合KCNQ1OT1。此外，KCNQ1OT1的下调通过上调CRC细胞中CTNND1的表达促进CRC进展。此外，在体内，KCNQ1OT1敲低通过miR-329-3p/CTNND1轴抑制CRC肿瘤生长。
我们的结果表明，KCNQ1OT1可通过海绵吸附miR-329-3p正向调节CTNND1的表达，从而促进CRC的进展。我们的研究结果为CRC提供了潜在的治疗靶点。
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