Department of Gynecology and Obstetrics, Beijing Tongren Hospital Affiliated to Capital Medical University, Beijing, China.
Neoplasma. 2020 Jul;67(4):782-793. doi: 10.4149/neo_2020_190617N515. Epub 2020 Apr 16.
Long noncoding RNAs (LncRNAs) show great potential as the therapeutic targets attributing to their implication in the progression of various human cancers, including ovarian cancer (OC). Here, we aimed to explore the biological function of lncRNA cyclin-dependent kinase inhibitor 2B antisense RNA 1 (CDKN2B-AS1) in OC and its mechanism of action. The abundances of CDKN2B-AS1, miR-143-3p, and SMAD3 mRNA were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell Counting Kit-8 (CCK8) was performed to analyze cell proliferation. Cell apoptosis was assessed by flow cytometry and western blot analyses. Transwell assay was utilized to analyze cell migration and invasion abilities. Tumor xenograft was performed to confirm the role of CDKN2B-AS1 in ovarian tumor growth in vivo. The protein level of SMAD3 was examined by western blot assay. The interaction between CDKN2B-AS1 and miR-143-3p, or miR-143-3p and SMAD3 was demonstrated by bioinformatic, luciferase reporter, qRT-PCR and western blot analyses. CDKN2B-AS1 was upregulated in OC and correlated with clinicopathologic features. The knockdown of CDKN2B-AS1 hampered the development of OC, as reflected by the suppression of cell proliferation, migration, and invasion, and the enhancement of cell apoptosis, whereas the effects could be rescued by the overexpression of SMAD3. The absence of CDKN2B-AS1 blocked tumor growth in vivo. CDKN2B-AS1 served as a molecular sponge for miR-143-3p, leading to the derepression of miR-143-3p target SMAD3, which eventually triggered the progression of OC. In conclusion, CDKN2B-AS1 promoted tumor growth, invasion, and migration of OC by regulation of miR-143-3p/SMAD3 axis, hinting that CDKN2B-AS1 might be a potential biomarker for OC diagnosis and treatment.
长链非编码 RNA（lncRNA）在各种人类癌症的进展中具有重要的治疗靶点潜力，包括卵巢癌（OC）。在这里，我们旨在探讨 lncRNA 细胞周期蛋白依赖性激酶抑制剂 2B 反义 RNA 1（CDKN2B-AS1）在 OC 中的生物学功能及其作用机制。通过实时定量聚合酶链反应（qRT-PCR）测定 CDKN2B-AS1、miR-143-3p 和 SMAD3 mRNA 的丰度。细胞计数试剂盒-8（CCK8）用于分析细胞增殖。通过流式细胞术和 Western blot 分析评估细胞凋亡。Transwell 测定用于分析细胞迁移和侵袭能力。肿瘤异种移植用于体内证实 CDKN2B-AS1 在卵巢肿瘤生长中的作用。Western blot 测定用于检测 SMAD3 蛋白水平。通过生物信息学、荧光素酶报告基因、qRT-PCR 和 Western blot 分析证实了 CDKN2B-AS1 与 miR-143-3p 或 miR-143-3p 与 SMAD3 之间的相互作用。CDKN2B-AS1 在 OC 中上调，并与临床病理特征相关。CDKN2B-AS1 的敲低阻碍了 OC 的发展，表现为细胞增殖、迁移和侵袭的抑制，细胞凋亡的增强，而 SMAD3 的过表达可以挽救这些作用。CDKN2B-AS1 的缺失阻断了体内肿瘤的生长。CDKN2B-AS1 作为 miR-143-3p 的分子海绵，导致 miR-143-3p 靶标 SMAD3 的去抑制，最终引发 OC 的进展。总之，CDKN2B-AS1 通过调节 miR-143-3p/SMAD3 轴促进 OC 的肿瘤生长、侵袭和迁移，提示 CDKN2B-AS1 可能是 OC 诊断和治疗的潜在生物标志物。
