Department of Respiratory Medicine, Yantai Yuhuangding Hospital, Yantai 264000, China.
Department of Respiratory Medicine, Yantai Muping District Chinese Medical Hospital, Yantai 264100, China.
Exp Biol Med (Maywood). 2021 Jan;246(2):142-152. doi: 10.1177/1535370220961013. Epub 2020 Oct 6.
The long non-coding RNA colon cancer-associated transcript 1 (CCAT1) has been investigated to involve in the progression of non-small cell lung cancer (NSCLC). Thus, this study aims to explore the detailed molecular mechanisms of CCAT1 in NSCLC. The expression of CCAT1, miR-216a-5p, RAP2B, Bax, Bcl-2, and cleaved caspase 3 was detected by qRT-PCR or Western blot. Cell proliferation, apoptosis, migration, and invasion were analyzed using cell counting kit-8, flow cytometry or Transwell assays, respectively. The interaction between miR-216a-5p and CCAT1 or RAP2B was analyzed by luciferase reporter, RNA immunoprecipitation, and pull-down assays. The expression of CCAT1 was elevated in NSCLC, and CCAT1 deletion could inhibit NSCLC cell proliferation, migration, and invasion but induce apoptosis as well as imped tumor growth . MiR-216a-5p was confirmed to be a target of CCAT1, and silencing miR-216a-5p could reverse CCAT1 depletion-mediated inhibitory effects on cell tumorigenesis in NSCLC. Besides that, miR-216a-5p was decreased in NSCLC, and miR-216a-5p restoration inhibited cell tumorigenesis by regulating RAP2B, which was verified to be a target of miR-216a-5p. Additionally, co-expression analysis suggested that CCAT1 indirectly regulated RAP2B level by targeting miR-216a-5p in NSCLC cells. Taken together, CCAT1 deletion could inhibit cell progression in NSCLC through miR-216a-5p/RAP2B axis, indicating a novel pathway underlying NSCLC cell progression and providing new potential targets for NSCLC treatment.
长链非编码 RNA 结肠癌相关转录本 1（CCAT1）已被研究涉及非小细胞肺癌（NSCLC）的进展。因此，本研究旨在探讨 CCAT1 在 NSCLC 中的详细分子机制。通过 qRT-PCR 或 Western blot 检测 CCAT1、miR-216a-5p、RAP2B、Bax、Bcl-2 和 cleaved caspase 3 的表达。分别使用细胞计数试剂盒-8、流式细胞术或 Transwell 测定法分析细胞增殖、凋亡、迁移和侵袭。通过荧光素酶报告、RNA 免疫沉淀和下拉测定分析 miR-216a-5p 与 CCAT1 或 RAP2B 的相互作用。CCAT1 在 NSCLC 中表达上调，CCAT1 缺失可抑制 NSCLC 细胞增殖、迁移和侵袭，但诱导凋亡并抑制肿瘤生长。MiR-216a-5p 被证实是 CCAT1 的靶标，沉默 miR-216a-5p 可逆转 CCAT1 缺失对 NSCLC 细胞肿瘤发生的抑制作用。此外，miR-216a-5p 在 NSCLC 中降低，miR-216a-5p 恢复通过调节 RAP2B 抑制细胞肿瘤发生，RAP2B 被证实是 miR-216a-5p 的靶标。此外，共表达分析表明，CCAT1 通过靶向 miR-216a-5p 在 NSCLC 细胞中间接调节 RAP2B 水平。总之，CCAT1 缺失可通过 miR-216a-5p/RAP2B 轴抑制 NSCLC 细胞进展，提示 NSCLC 细胞进展的新途径，并为 NSCLC 治疗提供新的潜在靶点。
