Department of Thoracic Surgery, the Third People's Hospital of Xinjiang Uygur Autonomous Region, Urumqi, Xinjiang, China.
Surgery Intensive Care Unit, Sichuan Academy of Medical Science & Sichuan Provincial People's Hospital, Chengdu, Qingyang, China.
Cell Cycle. 2022 Aug;21(16):1697-1709. doi: 10.1080/15384101.2022.2067438. Epub 2022 Apr 29.
Non-small cell lung cancer (NSCLC) is the most common lung cancer with a highest mortality rate. MiR-497-5p has been reported as tumor suppressor in many cancers, but the role and mechanism of miR-497-5p in regulating NSCLC progression are still largely unknown in vitro and in vivo. Here, miR-497-5p was significantly downregulated in human NSCLC tissues and cell lines, compared with matched adjacent tissues and normal lung epithelial cell line. Then, miR-497-5p mimic and inhibitor were, respectively, transfected into human NSCLC cells A549 and H460, CCK-8 assay, transwell assay, and flow cytometry were used to detect the capacities of cell proliferation, invasion and apoptosis. MiR-497-5p negatively regulated proliferation and invasion of NSCLC cancer cells. MiR-497-5p was demonstrated to directly bound to 3'-UTR of CX3CL1 mRNA and post-transcriptionally suppressed its expression thus inactivating its downstream oncogenic pathway ERK/AKT. Moreover, transfection with short hairpin RNA (shRNA) against CX3CL1 decreased capacity of cell proliferation and invasion and promoted cell apoptosis in NSCLC cells. In addition, ERK inhibitor U0126 attenuated the promotion effect of miR-497-5p inhibitor on activation of ERK/AKT and cell proliferation and migration. Finally, overexpression of miR-497-5p substantially suppressed activation of the ERK/AKT pathway and tumor growth in tumor-bearing mice in vivo. Taken together, our findings showed that miR-497-5p is downregulated in human NSCLC tissues and cell lines, and it inhibited tumor growth and cell invasion by targeting CX3CL1 gene to inactivate the ERK/AKT pathway in NSCLC cells.
非小细胞肺癌(NSCLC)是最常见的肺癌,死亡率最高。miR-497-5p 在许多癌症中被报道为肿瘤抑制因子,但在体外和体内,miR-497-5p 调节 NSCLC 进展的作用和机制在很大程度上仍然未知。在这里,与匹配的相邻组织和正常肺上皮细胞系相比,miR-497-5p 在人 NSCLC 组织和细胞系中显著下调。然后,miR-497-5p 模拟物和抑制剂分别转染入人 NSCLC 细胞 A549 和 H460,CCK-8 测定、Transwell 测定和流式细胞术用于检测细胞增殖、侵袭和凋亡的能力。miR-497-5p 负调节 NSCLC 癌细胞的增殖和侵袭。miR-497-5p 被证明直接结合 CX3CL1 mRNA 的 3'-UTR,并通过转录后抑制其表达从而使下游致癌通路 ERK/AKT 失活。此外,转染针对 CX3CL1 的短发夹 RNA (shRNA) 降低了 NSCLC 细胞的增殖和侵袭能力,并促进了细胞凋亡。此外,ERK 抑制剂 U0126 减弱了 miR-497-5p 抑制剂对 ERK/AKT 激活和细胞增殖和迁移的促进作用。最后,miR-497-5p 的过表达在体内肿瘤荷瘤小鼠中显著抑制了 ERK/AKT 通路的激活和肿瘤生长。总之,我们的研究结果表明,miR-497-5p 在人 NSCLC 组织和细胞系中下调,通过靶向 CX3CL1 基因抑制肿瘤生长和细胞侵袭,使 ERK/AKT 通路失活。