Department of Thoracic Surgery, The First Affiliated Hospital of Soochow University, Suzhou, China.
Department of Thoracic Surgery, The Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou, China.
Drug Deliv. 2022 Dec;29(1):1257-1271. doi: 10.1080/10717544.2022.2057617.
Circular RNAs (circRNAs) play important regulatory roles in multiple human malignancies, including non-small cell lung cancer (NSCLC). Here, we explored the role of circRNA vacuole membrane protein 1 (circVMP1) in NSCLC progression and cisplatin (DDP) resistance.
The DDP resistance, proliferation, sphere formation ability, migration, invasion, and apoptosis of NSCLC cells were analyzed by Cell Counting Kit-8 (CCK8) assay, 5-ethynyl-2'-deoxyuridine (EdU) assay, sphere formation assay, wound healing assay, Transwell assay, and flow cytometry. Methylated RIP-qPCR (MeRIP-qPCR) was conducted to analyze the mA modification level of SRY-box transcription factor 2 (SOX2). Dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, and RNA-pull down assay were performed to confirm the intermolecular interaction. Exosomes were identified by transmission electron microscopy (TEM) and characterized by nanoparticle tracking analysis (NTA).
CircVMP1 expression was markedly elevated in DDP-resistant NSCLC cell lines compared with their parental cell lines. CircVMP1 absence restrained the proliferation, sphere formation, migration, invasion, and DDP resistance and promoted the apoptosis of DDP-resistant NSCLC cells. CircVMP1 acted as microRNA-524-5p (miR-524-5p) sponge to up-regulate the expression of methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit (METTL3) and SOX2. CircVMP1 silencing restrained the malignant behaviors and DDP resistance of A549/DDP and H1299/DDP cells by targeting miR-524-5p. Exosomal circVMP1 disseminated the malignant properties and DDP resistance to DDP-sensitive cells. Exosomal circVMP1 elevated the DDP resistance of xenograft tumors . Exosomal circVMP1 was up-regulated in the serum samples of DDP-resistant NSCLC patients compared with DDP-sensitive patients.
Exosome-mediated transmission of circVMP1 promoted NSCLC progression and DDP resistance by targeting miR-524-5p-METTL3/SOX2 axis.HighlightsCircVMP1 level is up-regulated in DDP-resistant NSCLC cell lines compared with DDP-sensitive cell lines.CircVMP1 absence restrains the malignant behaviors and DDP resistance of A549/DDP and H1299/DDP cells.CircVMP1-miR-524-5p/METTL3/SOX2 axis is identified for the first time.CircVMP1 plays an oncogenic role by targeting miR-524-5p-METTL3/SOX2 axis in A549/DDP and H1299/DDP cells.Exosomal circVMP1 transmits the malignant properties and DDP resistance to DDP-sensitive cells.
环状 RNA（circRNAs）在多种人类恶性肿瘤中发挥重要的调控作用，包括非小细胞肺癌（NSCLC）。在这里，我们探讨了环状 RNA 液泡膜蛋白 1（circVMP1）在 NSCLC 进展和顺铂（DDP）耐药中的作用。
通过细胞计数试剂盒-8（CCK8）检测、5-乙炔基-2'-脱氧尿苷（EdU）检测、球体形成检测、划痕愈合检测、Transwell 检测和流式细胞术分析 NSCLC 细胞的 DDP 耐药性、增殖、球体形成能力、迁移、侵袭和凋亡。甲基化 RNA 免疫沉淀-qPCR（MeRIP-qPCR）分析性别决定区 Y 框转录因子 2（SOX2）的 mA 修饰水平。双荧光素酶报告基因检测、RNA 免疫沉淀（RIP）检测和 RNA 下拉检测证实了分子间的相互作用。通过透射电子显微镜（TEM）和纳米颗粒跟踪分析（NTA）鉴定外泌体。
与亲本细胞系相比，DDP 耐药性 NSCLC 细胞系中 circVMP1 的表达明显升高。circVMP1 的缺失抑制了 DDP 耐药性 NSCLC 细胞的增殖、球体形成、迁移、侵袭和 DDP 耐药性，并促进了细胞凋亡。circVMP1 作为 microRNA-524-5p（miR-524-5p）的海绵，上调了甲基转移酶 3、N6-腺苷甲基转移酶复合物催化亚基（METTL3）和 SOX2 的表达。circVMP1 沉默通过靶向 miR-524-5p 抑制了 A549/DDP 和 H1299/DDP 细胞的恶性行为和 DDP 耐药性。外泌体 circVMP1 通过靶向 miR-524-5p-METTL3/SOX2 轴将恶性表型和 DDP 耐药性传递给 DDP 敏感细胞。外泌体 circVMP1 提高了异种移植肿瘤的 DDP 耐药性。与 DDP 敏感患者相比，DDP 耐药性 NSCLC 患者的血清样本中 circVMP1 上调。
外泌体介导的 circVMP1 传递通过靶向 miR-524-5p-METTL3/SOX2 轴促进 NSCLC 的进展和 DDP 耐药性。
与 DDP 敏感细胞系相比，DDP 耐药性 NSCLC 细胞系中 circVMP1 的水平上调。circVMP1 的缺失抑制了 A549/DDP 和 H1299/DDP 细胞的恶性行为和 DDP 耐药性。首次鉴定了 circVMP1-miR-524-5p/METTL3/SOX2 轴。circVMP1 通过靶向 miR-524-5p-METTL3/SOX2 轴在 A549/DDP 和 H1299/DDP 细胞中发挥致癌作用。外泌体 circVMP1 通过靶向 miR-524-5p-METTL3/SOX2 轴将恶性表型和 DDP 耐药性传递给 DDP 敏感细胞。外泌体 circVMP1 提高了异种移植肿瘤的 DDP 耐药性。与 DDP 敏感患者相比，DDP 耐药性 NSCLC 患者的血清样本中 circVMP1 上调。
