Zhang Xiaomiao, Wang Jin, Pan Yuqin, Zhao Jun, Pan Yingge, Yan Yunqi, Shen Zhenya
Department of Cardiovascular Surgery, The First Affiliated Hospital of Soochow University, Suzhou 215006, P.R. China.
Department of Thoracic Surgery, Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 200080, P.R. China.
Oncol Lett. 2021 May;21(5):389. doi: 10.3892/ol.2021.12650. Epub 2021 Mar 17.
MicroRNA (miR)-365b-3p has been recently reported to induce cell cycle arrest and apoptosis in retinoblastoma; however, its expression pattern and biological function in non-small cell lung cancer (NSCLC) remain unknown. The present study aimed to investigate the functional role of miR-365b-3p in NSCLC. The results demonstrated that miR-365b-3p expression level was significantly decreased in NSCLC tissues and cell lines compared with controls using reverse transcriptase quantitative PCR. Furthermore, miR-365b-3p expression level was overexpressed by miR-365b-3p mimics transfection in A549 cells, whereas it was downregulated following H1299 cell transfection with miR-365b-3p inhibitor. Restoration of miR-365b-3p inhibited cell proliferation, induced cell cycle G0/G1 arrest and stimulated apoptosis in A549 cells using CCK-8 assay, colony formation and flow cytometry assay. However, miR-365b-3p inhibitor had the opposite effects in H1299 cells. Furthermore, results from bioinformatics analysis and luciferase reporter assay confirmed that serine/threonine protein phosphatase 5 (PPP5C) was a direct target of miR-365b-3p. In addition, online Kaplan-Meier plotter software demonstrated that high PPP5C expression level was associated with lower overall survival and disease-free survival in patients with NSCLC. Furthermore, PPP5C knockdown imitated the effects of miR-365b-3p mimics on A549 cell proliferation, cell cycle distribution and apoptosis, whereas its overexpression rescued the effects of miR-365b-3p mimics on A549 cell proliferation, cell cycle distribution and apoptosis. In conclusion, the findings from the present study suggested that miR-365b-3p may partly suppress NSCLC cell behaviors by targeting PPP5C, which may represent a promising therapeutic target for patients with NSCLC.
最近有报道称，微小RNA（miR）-365b-3p可诱导视网膜母细胞瘤细胞周期停滞和凋亡；然而，其在非小细胞肺癌（NSCLC）中的表达模式和生物学功能尚不清楚。本研究旨在探讨miR-365b-3p在NSCLC中的功能作用。结果表明，与对照组相比，使用逆转录定量PCR检测发现NSCLC组织和细胞系中miR-365b-3p表达水平显著降低。此外，通过转染miR-365b-3p模拟物使A549细胞中miR-365b-3p表达水平上调，而用miR-365b-3p抑制剂转染H1299细胞后其表达水平下调。使用CCK-8检测、集落形成实验和流式细胞术分析，恢复miR-365b-3p可抑制A549细胞增殖，诱导细胞周期G0/G1期停滞并刺激细胞凋亡。然而，miR-365b-3p抑制剂在H1299细胞中具有相反的作用。此外，生物信息学分析和荧光素酶报告基因检测结果证实，丝氨酸/苏氨酸蛋白磷酸酶5（PPP5C）是miR-365b-3p的直接靶点。此外，在线Kaplan-Meier Plotter软件显示，PPP5C高表达水平与NSCLC患者较低的总生存期和无病生存期相关。此外，敲低PPP5C可模拟miR-365b-3p模拟物对A549细胞增殖、细胞周期分布和凋亡的影响，而其过表达可挽救miR-365b-3p模拟物对A549细胞增殖、细胞周期分布和凋亡的影响。总之，本研究结果表明，miR-365b-3p可能通过靶向PPP5C部分抑制NSCLC细胞行为，这可能是NSCLC患者一个有前景的治疗靶点。
