Department of Dermatology, General Hospital of Ningxia Medical University, Yinchuan City, China.
Department of Pathology, School of Basic Medicine, Ningxia Medical University, Yinchuan City, China.
Bioengineered. 2022 Jan;13(1):1209-1223. doi: 10.1080/21655979.2021.2019172.
Circular RNAs (circRNAs) have shown pivotal regulatory roles in tumorigenesis and progression. Our purpose was to analyze the role of circRNA La ribonucleoprotein 1B (circ-LARP1B; hsa_circ_0070934) in cutaneous squamous cell carcinoma (CSCC) progression and its associated mechanism. Cell viability, colony formation ability, migration, and invasion were analyzed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide (MTT) assay, colony formation assay, wound healing assay, and transwell invasion assay. Flow cytometry was performed to analyze cell apoptosis and cell cycle progression. Cell glycolytic metabolism was analyzed using Glucose Uptake Colorimetric Assay kit, Lactate Assay Kit II, and ATP colorimetric Assay kit. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to verify the interaction between microRNA-515-5p (miR-515-5p) and circ-LARP1B or TPX2 microtubule nucleation factor (TPX2). Circ-LARP1B expression was up-regulated in CSCC tissues and cell lines. Circ-LARP1B knockdown suppressed cell viability, colony formation ability, migration, invasion, cell cycle progression, and glycolysis and triggered cell apoptosis in CSCC cells. miR-515-5p was a direct target of circ-LARP1B in CSCC cells, and circ-LARP1B silencing-mediated anti-tumor effects were largely counteracted by miR-515-5p knockdown. miR-515-5p directly interacted with the 3' untranslated region (3'UTR) of TPX2. TPX2 overexpression largely overturned miR-515-5p-mediated anti-tumor effects in CSCC cells. Circ-LARP1B could up-regulate TPX2 expression by sponging miR-515-5p in CSCC cells. Circ-LARP1B knockdown suppressed tumor growth . In conclusion, circ-LARP1B contributed to CSCC progression by targeting miR-515-5p/TPX2 axis. The circ-LARP1B/miR-515-5p/TPX2 axis might provide novel therapeutic targets for CSCC patients.
环状 RNA(circRNA)在肿瘤发生和进展中发挥着关键的调节作用。我们的目的是分析环状 RNA La 核蛋白 1B(circ-LARP1B;hsa_circ_0070934)在皮肤鳞状细胞癌(CSCC)进展中的作用及其相关机制。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)测定法、集落形成测定法、划痕愈合测定法和 Transwell 侵袭测定法分析细胞活力、集落形成能力、迁移和侵袭。通过流式细胞术分析细胞凋亡和细胞周期进程。使用葡萄糖摄取比色测定试剂盒、乳酸测定试剂盒 II 和 ATP 比色测定试剂盒分析细胞糖酵解代谢。通过双荧光素酶报告基因测定和 RNA 免疫沉淀(RIP)测定验证 microRNA-515-5p(miR-515-5p)与 circ-LARP1B 或微管成核因子 TPX2(TPX2)之间的相互作用。circ-LARP1B 在 CSCC 组织和细胞系中表达上调。circ-LARP1B 敲低抑制 CSCC 细胞的细胞活力、集落形成能力、迁移、侵袭、细胞周期进程和糖酵解,并触发细胞凋亡。miR-515-5p 是 CSCC 细胞中 circ-LARP1B 的直接靶标,circ-LARP1B 沉默介导的抗肿瘤作用在很大程度上被 miR-515-5p 敲低所抵消。miR-515-5p 直接与 TPX2 的 3'非翻译区(3'UTR)相互作用。TPX2 过表达在 CSCC 细胞中很大程度上推翻了 miR-515-5p 介导的抗肿瘤作用。circ-LARP1B 可通过海绵吸附 miR-515-5p 在 CSCC 细胞中上调 TPX2 表达。circ-LARP1B 敲低抑制肿瘤生长。总之,circ-LARP1B 通过靶向 miR-515-5p/TPX2 轴促进 CSCC 进展。circ-LARP1B/miR-515-5p/TPX2 轴可能为 CSCC 患者提供新的治疗靶点。