Department of Neurosurgery, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou 563000, People's Republic of China.
Int J Nanomedicine. 2020 Nov 10;15:8863-8872. doi: 10.2147/IJN.S271795. eCollection 2020.
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.
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.
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.
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.
神经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，神经胶质瘤的临床治疗方法有限。通过基因治疗为神经胶质瘤寻求新的治疗方法具有重要意义。
使用微阵列鉴定神经胶质瘤中差异表达的 lncRNA。通过 qRT-PCR 检测长非编码 RNA（lncRNA）ROR1-AS1 和 miR-4686 的表达。从正常和癌细胞的上清液中分离外泌体，并用 TEM 进行外泌体鉴定。MTT 测定、划痕愈合测定、Transwell 测定和集落形成测定用于检测外泌体 ROR1-AS1 对神经胶质瘤细胞增殖、迁移和侵袭的功能。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。通过裸鼠异种移植模型证实外泌体 ROR1-AS1 对神经胶质瘤肿瘤发生的影响。
ROR1-AS1 在神经胶质瘤组织中上调，ROR1-AS1 的高表达表明神经胶质瘤患者预后不良。有趣的是，ROR1-AS1 被包装到外泌体中并源自肿瘤细胞。功能分析表明外泌体 ROR1-AS1 促进了神经胶质瘤细胞系 SHG44 和 U251 的进展。此外，ROR1-AS1 作为 miR-4686 的海绵并抑制其表达。功能上，miR-4686 的强制表达消除了 lncRNA ROR1-AS1 对神经胶质瘤发展的促进作用。体内肿瘤发生实验表明，外泌体 ROR1-AS1 通过 miR-4686 轴促进神经胶质瘤的发展。
我们的研究表明，肿瘤细胞衍生的外泌体 ROR1-AS1 通过抑制 miR-4686 促进神经胶质瘤进展，这可能是神经胶质瘤临床治疗的潜在治疗靶点。
