METHODS: Serum exosomes were isolated from both SCLC patients and healthy controls, and subjected to exosomal miRNA sequencing analysis. Mimics and inhibitors were employed to investigate the function of exosomal miR-1128-5p in cell migration and proliferation, both in vitro and in vivo. Western blot and luciferase assay were utilized to identify the interaction between miR-1228-5p and dual specificity phosphatase 22 (DUSP22).
RESULTS: Exosomal miRNA sequencing analysis revealed enrichment of specific miRNAs in SCLC compared to healthy controls. Circulating miR-1228-5p was upregulated in SCLC patients, associated with advanced stages, suggesting its potential oncogenic role. In vitro, miR-1228-5p expression was significantly higher in SCLC cells than in normal cells. SCLC cell-derived exosomes contained elevated levels of miR-1228-5p, facilitating its entry into co-cultured cells. Notably, migration and proliferation induced by SCLC exosomes were mainly mediated by miR-1228-5p. In vivo experiments confirmed these findings. Western blot analysis demonstrated miR-1228-5p\'s regulation of DUSP22 expression, and luciferase reporter assay validated DUSP22 as a direct target gene. Overexpressing DUSP22 counteracted miR-1228-5p\'s promotion of SCLC cell proliferation and migration.
CONCLUSIONS: Collectively, our results suggest that exosomes play a role in facilitating cancer growth and metastasis by delivering miR-1228-5p. Moreover, circulating exosomal miR-1228-5p may serve as a potential marker for SCLC diagnosis and prognosis.
方法:从SCLC患者和健康对照中分离血清外泌体,并进行外泌体miRNA测序分析。模拟和抑制剂用于研究外泌体miR-1128-5p在细胞迁移和增殖中的功能。在体外和体内。使用蛋白质印迹和荧光素酶测定来鉴定miR-1228-5p与双特异性磷酸酶22(DUSP22)之间的相互作用。
结果:外泌体miRNA测序分析显示,与健康对照相比,SCLC中特定miRNA的富集。循环miR-1228-5p在SCLC患者中上调,与高级阶段相关,提示其潜在的致癌作用。体外,miR-1228-5p在SCLC细胞中的表达显著高于正常细胞。SCLC细胞来源的外泌体含有升高水平的miR-1228-5p,促进其进入共培养细胞。值得注意的是,SCLC外泌体诱导的细胞迁移和增殖主要由miR-1228-5p介导。体内实验证实了这些发现。Westernblot分析显示miR-1228-5p调节DUSP22表达,和荧光素酶报告基因分析验证了DUSP22作为直接靶基因。过表达DUSP22抑制miR-1228-5p促进SCLC细胞增殖和迁移。
结论:总的来说,我们的结果表明,外泌体通过递送miR-1228-5p在促进癌症生长和转移中发挥作用.此外,循环外泌体miR-1228-5p可作为SCLC诊断和预后的潜在标志物.