Long noncoding RNAs (lncRNAs) exert crucial effects on the development of many malignancies, including gastric cancer. Herein, we investigated the role of lncRNA noncatalytic region of tyrosine kinase adaptor protein 1 (NCK1) divergent transcript (NCK1-DT, also known as NCK1-AS1) in gastric cancer. Reverse transcription quantitative polymerase chain reaction demonstrated that NCK1-AS1 exhibited high expression in gastric cancer tissues and cells. In vitro assays including MTT, colony formation, Transwell, wound healing and sphere formation assays indicated that NCK1-AS1 depletion inhibited cell proliferation, migration, invasion and stemness maintenance. Luciferase reporter and RIP assays suggested that NCK1-AS1 functioned as a competitive endogenous RNA (ceRNA) for miR-22-3p to positively modulate BCL9 expression. BCL9 was a target gene of miR-22-3p. According to western blot analysis and TOP/FOP flash assay, NCK1-AS1 activated the Wnt/β-catenin signaling via the miR-22-3p/BCL9 axis. Furthermore, rescue experiments verified that NCK1-AS1 affected cellular processes by activating the Wnt/β-catenin signaling pathway via the miR-22-3p/BCL9 axis. Tumor xenograft model validated that NCK1-AS1 promoted tumor growth in vivo via the Wnt/β-catenin signaling by upregulating BCL9 expression. Overall, NCK1-AS1 functions as an oncogene and promotes gastric cancer progression via the miR-22-3p/BCL9-Wnt/β-catenin signaling pathway.

中文翻译：

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LncRNA NCK1-AS1 通过靶向 miR-22-3p/BCL9 轴激活 Wnt/β-catenin 信号通路在胃癌中发挥致癌作用

长链非编码 RNA (lncRNA) 对包括胃癌在内的许多恶性肿瘤的发展发挥着至关重要的作用。在此，我们研究了酪氨酸激酶衔接蛋白 1 (NCK1) 不同转录本 (NCK1-DT，也称为 NCK1-AS1) 的 lncRNA 非催化区在胃癌中的作用。逆转录定量聚合酶链反应表明NCK1-AS1在胃癌组织和细胞中高表达。包括 MTT、集落形成、Transwell、伤口愈合和球体形成测定在内的体外测定表明，NCK1-AS1 消耗抑制细胞增殖、迁移、侵袭和干性维持。荧光素酶报告基因和 RIP 分析表明 NCK1-AS1 作为 miR-22-3p 的竞争性内源性 RNA (ceRNA) 发挥正向调节 BCL9 表达的作用。BCL9 是 miR-22-3p 的靶基因。根据蛋白质印迹分析和 TOP/FOP 闪存分析，NCK1-AS1 通过 miR-22-3p/BCL9 轴激活 Wnt/β-catenin 信号。此外，救援实验证实 NCK1-AS1 通过 miR-22-3p/BCL9 轴激活 Wnt/β-catenin 信号通路影响细胞过程。肿瘤异种移植模型验证 NCK1-AS1 通过上调 BCL9 表达通过 Wnt/β-catenin 信号传导促进体内肿瘤生长。总体而言，NCK1-AS1 作为癌基因发挥作用，通过 miR-22-3p/BCL9-Wnt/β-catenin 信号通路促进胃癌进展。救援实验证实 NCK1-AS1 通过 miR-22-3p/BCL9 轴激活 Wnt/β-catenin 信号通路影响细胞过程。肿瘤异种移植模型验证 NCK1-AS1 通过上调 BCL9 表达通过 Wnt/β-catenin 信号传导促进体内肿瘤生长。总体而言，NCK1-AS1 作为癌基因发挥作用，通过 miR-22-3p/BCL9-Wnt/β-catenin 信号通路促进胃癌进展。救援实验证实 NCK1-AS1 通过 miR-22-3p/BCL9 轴激活 Wnt/β-catenin 信号通路影响细胞过程。肿瘤异种移植模型验证 NCK1-AS1 通过上调 BCL9 表达通过 Wnt/β-catenin 信号传导促进体内肿瘤生长。总体而言，NCK1-AS1 作为癌基因发挥作用，通过 miR-22-3p/BCL9-Wnt/β-catenin 信号通路促进胃癌进展。