Objective: Glioma is one of the most common central nervous system malignant tumors, accounting for 45%– 60% of adult intracranial tumors. However, the clinical treatment of glioma is limited. It is of great significance to seek new therapeutic methods for glioma via gene therapy.
Materials and Methods: Microarray is used to identify the lncRNAs that are differentially expressed in glioma. The expression of long non-coding RNA (lncRNA) ROR1-AS1 and miR-4686 was detected by qRT-PCR. Exosomes were isolated from the supernatant of normal and cancerous cells, and TEM was used for exosomes identification. MTT assay, wound healing assay, transwell assay, and colony formation assay were used to detect the exo-ROR1-AS1 function on proliferation, migration, and invasion in glioma cells. Luciferase assay and RIP assay were used to identify the relationship between lncRNA ROR1-AS1 and miR-4686. The effect of exo-ROR1-AS1 on tumorigenesis of glioma was confirmed by the xenograft nude mice model.
Results: ROR1-AS1 was up-regulated in glioma tissues, and the high expression of ROR1-AS1 indicated a poor prognosis in glioma patients. Interestingly, ROR1-AS1 was packaged into exosomes and derived from tumor cells. Functional analysis showed exo-ROR1-AS1 promoted the progression of glioma cell lines SHG44 and U251. Furthermore, ROR1-AS1 acted as a sponge of miR-4686 and inhibited its expression. Functionally, forced expression of miR-4686 removed the promoted effects of lncRNA ROR1-AS1 on glioma development. In vivo tumorigenesis experiments showed that exo-ROR1-AS1 promoted glioma development via miR-4686 axis.
Conclusion: Our study suggested tumor cells derived exo-ROR1-AS1 promoted glioma progression by inhibiting miR-4686, which might be a potential therapeutic target for glioma clinical treatment.

Keywords: glioma, lncRNA ROR1-AS1, exosome, miR-4686, tumorigenesis


中文翻译：

---

来自肿瘤细胞的外泌体lncRNA ROR1-AS1通过调节miR-4686促进胶质瘤进展

目的：胶质瘤是最常见的中枢神经系统恶性肿瘤之一，占成人颅内肿瘤的45%~60%。然而，胶质瘤的临床治疗是有限的。通过基因治疗寻求新的胶质瘤治疗方法具有重要意义。
材料和方法：微阵列用于识别在胶质瘤中差异表达的 lncRNA。通过qRT-PCR检测长链非编码RNA（lncRNA）ROR1-AS1和miR-4686的表达。从正常和癌细胞的上清液中分离出外泌体，并使用TEM进行外泌体鉴定。采用MTT法、伤口愈合法、transwell法和集落形成法检测exo-ROR1-AS1对胶质瘤细胞增殖、迁移和侵袭的作用。荧光素酶测定和RIP测定用于鉴定lncRNA ROR1-AS1和miR-4686之间的关系。异种移植裸鼠模型证实了exo-ROR1-AS1对胶质瘤肿瘤发生的影响。
结果：ROR1-AS1在胶质瘤组织中上调，ROR1-AS1的高表达表明胶质瘤患者预后不良。有趣的是，ROR1-AS1 被包装到外泌体中并来源于肿瘤细胞。功能分析显示 exo-ROR1-AS1 促进了胶质瘤细胞系 SHG44 和 U251 的进展。此外，ROR1-AS1 充当 miR-4686 的海绵并抑制其表达。在功能上，miR-4686 的强制表达消除了 lncRNA ROR1-AS1 对胶质瘤发展的促进作用。体内肿瘤发生实验表明，exo-ROR1-AS1 通过 miR-4686 轴促进神经胶质瘤的发展。
结论：我们的研究表明，exo-ROR1-AS1 衍生的肿瘤细胞通过抑制 miR-4686 促进胶质瘤进展，这可能是胶质瘤临床治疗的潜在治疗靶点。

关键词：胶质瘤，lncRNA ROR1-AS1，外泌体，miR-4686，肿瘤发生