Department of Respiratory and Critical Care Medicine, The Affiliated Hospital of Northwest University, Xi'an No.3 Hospital, Xi'an 710018, China.
Department of Respiratory and Critical Care Medicine, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, China.
Acta Biochim Biophys Sin (Shanghai). 2021 Feb 4;53(2):189-200. doi: 10.1093/abbs/gmaa159.
miR-195-5p has been widely explored in various cancers and is considered as a tumor-suppressive microRNA. However, its roles in human lung cancer pathogenesis are not fully elucidated. In this study, we aimed to explore how miR-195-5p is involved in malignant behaviors of lung adenocarcinoma (LUAD) cells. miR-195-5p expression was examined in the tumor tissues of patients with LUAD and human LUAD cell lines including A549 and PC-9. Thioredoxin reductase 2 (TrxR2) was predicted to be an mRNA target of miR-195-5p using online tools and validated by the Dual-Luciferase Reporter Assay. Lentivirus infection was used for gene overexpression, while gene knockdown was achieved by RNA interference. Cell proliferation was determined by Cell Counting Kit-8 and 5-ethynyl-2'-deoxyuridine methods, and cell migration and invasion were assayed with transwell experiments. Cell apoptosis was determined by annexin V staining-based flow cytometry. The antitumor effects of miR-195-5p were also evaluated in nude mice xenografted with A549 cells. We found that miR-195-5p was lowly expressed in human LUAD cells, and its overexpression markedly suppressed cell proliferation, migration, and invasion and increased the apoptosis of LUAD cells in vitro. TrxR2 knockdown phenocopied the tumor-suppressive effects of miR-195-5p overexpression, while simultaneous TrxR2 overexpression remarkably reversed the effects of miR-195-5p overexpression on malignant behaviors of A549 and PC-9 cells. Additionally, miR-195-5p overexpression inhibited the growth of xenografted A549 tumor in nude mice. Our work verified that miR-195-5p exerts tumor-suppressive functions in LUAD cells through targeting TrxR2 and suggested that the miR-195-5p/TrxR2 axis is a potential biomarker for LUAD therapy.
miR-195-5p 在各种癌症中广泛研究,被认为是一种肿瘤抑制性 microRNA。然而,其在人类肺癌发病机制中的作用尚未完全阐明。在本研究中,我们旨在探讨 miR-195-5p 如何参与肺腺癌(LUAD)细胞的恶性行为。检测了 LUAD 患者肿瘤组织和 A549、PC-9 等人类 LUAD 细胞系中 miR-195-5p 的表达。使用在线工具预测硫氧还蛋白还原酶 2(TrxR2)是 miR-195-5p 的 mRNA 靶标,并通过双荧光素酶报告基因检测进行验证。慢病毒感染用于基因过表达,而 RNA 干扰用于基因敲低。通过细胞计数试剂盒-8 和 5-乙炔基-2'-脱氧尿苷法测定细胞增殖,通过 Transwell 实验测定细胞迁移和侵袭,通过 Annexin V 染色流式细胞术测定细胞凋亡。还在 A549 细胞裸鼠异种移植模型中评估了 miR-195-5p 的抗肿瘤作用。结果发现,miR-195-5p 在人类 LUAD 细胞中低表达,其过表达显著抑制 LUAD 细胞的增殖、迁移和侵袭,并增加体外细胞凋亡。TrxR2 敲低模拟了 miR-195-5p 过表达的肿瘤抑制作用,而同时过表达 TrxR2 则显著逆转了 miR-195-5p 过表达对 A549 和 PC-9 细胞恶性行为的影响。此外,miR-195-5p 过表达抑制了裸鼠异种移植 A549 肿瘤的生长。本研究验证了 miR-195-5p 通过靶向 TrxR2 发挥对 LUAD 细胞的肿瘤抑制作用,并提示 miR-195-5p/TrxR2 轴可能是 LUAD 治疗的潜在生物标志物。