a Department of Gastroenterology , the Second Affiliated Hospital of Xi'an Jiaotong University , Xi'an , China.
b Department of Gastroenterology, Shenzhen Sixth People's Hospital (Nanshan Hospital) , Huazhong University of Science and Technology Union Shenzhen Hospital , Shenzhen , China.
Cell Cycle. 2018;17(15):1886-1900. doi: 10.1080/15384101.2018.1502574. Epub 2018 Sep 10.
In this study, we aimed to investigate the effects of lncRNA CASC11 on gastric cancer (GC) cell progression through regulating miR-340-5p and cell cycle pathway. Expressions of lncRNA CASC11 in gastric cancer tissues and cell lines were determined by qRT-PCR. Differentially expressed lncRNAs, mRNAs and miRNAs were screened through microarray analysis. The relationship among CASC11, CDK1 and miR-340-5p was predicted by TargetScan and validated through dual luciferase reporter assay. Western blot assay examined the protein level of CDK1 and several cell cycle regulatory proteins. GO functional analysis and KEGG pathway analysis were used to predict the association between functions and related pathways. Cell proliferation was determined by CCK-8 assays. Cell apoptosis and cell cycle were detected by flow cytometry assay. CASC11 was highly expressed in GC tissues and cell lines. Knockdown of CASC11 inhibited GC cell proliferation, promoted cell apoptosis and blocked cell cycle. KEGG further indicated an enriched cell cycle pathway involving CDK1. QRT-PCR showed that miR-340-5p was down-regulated in GC cells tissues, while CDK1 was up-regulated. Furthermore, CASC11 acted as a sponge of miR-340-5p which directly targeted CDK1. Meanwhile, miR-340-5p overexpression promoted GC cell apoptosis and induced cell cycle arrest, while CDK1 overexpression inhibited cell apoptosis and accelerated cell cycle. Our study revealed the mechanism of CASC11/miR-340-5p/CDK1 network in GC cell line, and suggested that CASC11 was a novel facilitator that exerted a biological effect by activating the cell cycle signaling pathway. This finding provides a potential therapeutic target for GC.
在这项研究中，我们旨在通过调节 miR-340-5p 和细胞周期途径来研究 lncRNA CASC11 对胃癌（GC）细胞进展的影响。通过 qRT-PCR 测定胃癌组织和细胞系中 lncRNA CASC11 的表达。通过微阵列分析筛选差异表达的 lncRNA、mRNA 和 miRNA。通过 TargetScan 预测 CASC11、CDK1 和 miR-340-5p 之间的关系，并通过双荧光素酶报告基因测定验证。Western blot 测定检测 CDK1 和几个细胞周期调节蛋白的蛋白水平。GO 功能分析和 KEGG 途径分析用于预测功能与相关途径之间的关联。通过 CCK-8 测定法测定细胞增殖。通过流式细胞术检测细胞凋亡和细胞周期。CASC11 在 GC 组织和细胞系中高表达。敲低 CASC11 抑制 GC 细胞增殖，促进细胞凋亡并阻断细胞周期。KEGG 进一步表明涉及 CDK1 的富含细胞周期途径得到了富集。QRT-PCR 显示 miR-340-5p 在 GC 细胞组织中下调，而 CDK1 上调。此外，CASC11 作为 miR-340-5p 的海绵，直接靶向 CDK1。同时，miR-340-5p 过表达促进 GC 细胞凋亡并诱导细胞周期停滞，而 CDK1 过表达抑制细胞凋亡并加速细胞周期。本研究揭示了 CASC11/miR-340-5p/CDK1 网络在 GC 细胞系中的作用机制，并表明 CASC11 是一种通过激活细胞周期信号通路发挥生物学作用的新型促进剂。这一发现为 GC 提供了一个潜在的治疗靶点。
