Department of Cardiothoracic Surgery, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, Jingzhou, Hubei 434000, P.R. China.
Mol Med Rep. 2020 Jun;21(6):2589-2596. doi: 10.3892/mmr.2020.11033. Epub 2020 Mar 20.
Non-small-cell lung cancer (NSCLC) is the fundamental form of lung cancer and the leading cause of cancer‑related mortality in humans. Numerous studies have identified a role for microRNAs (miRs) in cell proliferation, invasion and metastasis in numerous types of cancer, including lung cancer. In the present study, the functional roles and molecular mechanisms of miR‑28 in NSCLC tumorigenesis were investigated. Reverse transcription‑quantitative PCR (RT‑qPCR) was used to measure miR‑28 expression levels in NSCLC tumor tissues and cell lines. A dual‑luciferase assay was performed to observe the direct interaction between miR‑28 and PTEN in A549 cells. Furthermore, the effect of miR‑28 on the mRNA and protein expression levels of PTEN was examined by RT‑qPCR and western blotting, respectively. A Cell Counting kit‑8 assay was performed to identify the relationship between the miR‑28/PTEN axis and tumor cell proliferation using cells infected with lentivirus (LV)‑anti‑miR‑28 or LV‑anti‑miR‑28 + short hairpin RNA‑PTEN. miR‑28 expression was upregulated in NSCLC tumor tissues and cell lines compared with the control groups. PTEN was identified as the downstream gene of miR‑28 in NSCLC and was negatively regulated by miR‑28. In addition, miR‑28 knockdown suppressed the proliferation of A549 and H292 cells. Cells infected with LV‑anti‑miR‑28 + short hairpin RNA‑PTEN promoted tumor cell proliferation in A549 and H292 cells compared with cells infected with LV‑anti‑miR‑28. Taken together, the present study suggested that miR‑28 might serve as the promoter in the development of NSCLC by targeting PTEN. Therefore, the miR‑28/PTEN axis may serve as a potential diagnostic and therapeutic target for NSCLC.
非小细胞肺癌(NSCLC)是肺癌的基本形式,也是人类癌症相关死亡的主要原因。许多研究已经确定了 microRNAs(miRs)在包括肺癌在内的多种癌症中的细胞增殖、侵袭和转移中的作用。在本研究中,研究了 miR-28 在 NSCLC 肿瘤发生中的功能作用和分子机制。逆转录-定量 PCR(RT-qPCR)用于测量 NSCLC 肿瘤组织和细胞系中的 miR-28 表达水平。双荧光素酶报告基因实验观察 miR-28 在 A549 细胞中与 PTEN 的直接相互作用。此外,通过 RT-qPCR 和 Western blot 分别检测 miR-28 对 PTEN 的 mRNA 和蛋白表达水平的影响。细胞计数试剂盒-8 检测感染慢病毒(LV)-anti-miR-28 或 LV-anti-miR-28+短发夹 RNA-PTEN 的细胞中 miR-28/PTEN 轴与肿瘤细胞增殖的关系。与对照组相比,miR-28 在 NSCLC 肿瘤组织和细胞系中上调。PTEN 被鉴定为 NSCLC 中 miR-28 的下游基因,并受 miR-28 负调控。此外,miR-28 敲低抑制了 A549 和 H292 细胞的增殖。与感染 LV-anti-miR-28 的细胞相比,感染 LV-anti-miR-28+短发夹 RNA-PTEN 的细胞促进了 A549 和 H292 细胞中的肿瘤细胞增殖。总之,本研究表明,miR-28 可能通过靶向 PTEN 作为 NSCLC 发展的促进因子。因此,miR-28/PTEN 轴可能成为 NSCLC 的潜在诊断和治疗靶点。