Department of Respiration, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Oncol Rep. 2020 Sep;44(3):1025-1036. doi: 10.3892/or.2020.7685. Epub 2020 Jul 13.
Emerging evidence has demonstrated that abnormally expressed long non‑coding (lnc) RNAs contribute to drug resistance in various types of malignancy. LncRNA antisense non‑coding RNA in the inhibitor of cyclin‑dependent kinase 4 locus (ANRIL) exerts oncogenic activity and acts as a key player in a variety of carcinomas, including non‑small cell lung cancer. The present study aimed to investigate the functional roles of ANRIL in cisplatin (DDP) resistance of lung cancer and the underlying mechanism involved in the competing endogenous RNA regulatory network. The expression levels of ANRIL and microRNA (miR)‑98 in lung cancer tissues and DDP‑resistant lung cancer cells were assessed by reverse transcription‑quantitative (RT‑q) PCR. The Pearson's correlation coefficient was used to determine the correlation between the expression levels of ANRIL and miR‑98 in lung cancer tissues. Cell proliferation and apoptosis were detected by MTT assay and flow cytometry analysis, respectively. The expression levels of proliferation‑ and apoptosis‑associated proteins were detected by western blot analysis. The potential interaction between ANRIL and miR‑98 was confirmed by RNA immunoprecipitation (RIP), dual luciferase reporter assay and RT‑qPCR. The results of the present study demonstrated that ANRIL was upregulated and miR‑98 was downregulated in lung cancer tissues and A549/DDP cells. In addition, the Pearson's correlation analysis revealed that the expression of ANRIL was significantly negatively correlated with that of miR‑98 in lung cancer tissues. ANRIL overexpression reversed the DDP‑induced cell proliferation suppression and apoptosis in lung cancer cells, whereas ANRIL knockdown exhibited the opposite effects. RIP, dual luciferase reporter assay and RT‑qPCR analysis results demonstrated that ANRIL directly interacted with miR‑98 and suppressed miR‑98 expression. Furthermore, transfection with the miR‑98 mimics promoted DDP‑induced cell proliferation inhibition and apoptosis in lung cancer cells, and these effects were partially reversed by ANRIL overexpression. In conclusion, ANRIL knockdown inhibited the development of DDP resistance by upregulating miR‑98, providing novel insights into the molecular mechanism of ANRIL involvement in DDP resistance of lung cancer cells.
越来越多的证据表明，异常表达的长非编码 (lnc) RNA 有助于各种恶性肿瘤的耐药性。位于细胞周期蛋白依赖性激酶 4 基因座的反义非编码 RNA 抑制因子 (ANRIL) 具有致癌活性，是多种癌症（包括非小细胞肺癌）的关键参与者。本研究旨在探讨 ANRIL 在肺癌顺铂 (DDP) 耐药中的功能作用及其在竞争内源性 RNA 调控网络中涉及的潜在机制。通过逆转录-定量 (RT-q) PCR 评估肺癌组织和 DDP 耐药肺癌细胞中 ANRIL 和 microRNA (miR)-98 的表达水平。采用 Pearson 相关系数分析肺癌组织中 ANRIL 和 miR-98 表达水平的相关性。通过 MTT 检测和流式细胞术分析分别检测细胞增殖和凋亡。通过 Western blot 分析检测增殖和凋亡相关蛋白的表达水平。通过 RNA 免疫沉淀 (RIP)、双荧光素酶报告基因检测和 RT-qPCR 验证 ANRIL 与 miR-98 之间的潜在相互作用。本研究结果表明，ANRIL 在肺癌组织和 A549/DDP 细胞中上调，miR-98 下调。此外，Pearson 相关分析显示，肺癌组织中 ANRIL 的表达与 miR-98 的表达呈显著负相关。过表达 ANRIL 逆转了 DDP 诱导的肺癌细胞增殖抑制和凋亡，而敲低 ANRIL 则呈现相反的效果。RIP、双荧光素酶报告基因检测和 RT-qPCR 分析结果表明，ANRIL 直接与 miR-98 相互作用并抑制 miR-98 的表达。此外，转染 miR-98 模拟物促进了 DDP 诱导的肺癌细胞增殖抑制和凋亡，而过表达 ANRIL 部分逆转了这些作用。综上所述，敲低 ANRIL 通过上调 miR-98 抑制 DDP 耐药的发展，为 ANRIL 参与肺癌细胞 DDP 耐药的分子机制提供了新的见解。
