Department of Neurology, The First Affiliated Hospital of Jinan University, Jinan University, Guangzhou, China.
Department of Otolaryngology, Beijing Yanqing District Hospital, Beijing, China.
Metab Brain Dis. 2022 Oct;37(7):2349-2362. doi: 10.1007/s11011-022-01034-0. Epub 2022 Jul 2.
The competing endogenous RNA (ceRNA) activity of long non-coding RNAs (lncRNAs) has profound effects in pathological disorders, including Parkinson's disease. Here, we focused on the LINC00943-mediated ceRNA network for the regulation of LINC00943 in MPP toxicity in SK-N-SH cells. SK-N-SH cells were exposed to 1-methyl-4-phenylpyridinium (MPP). LINC00943, miR-671-5p and ELAV like RNA binding protein 1 (ELAVL1) were quantified by real-time quantitative PCR (RT-qPCR) or western blot. Cell viability and apoptosis were gauged by Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. Direct relationship between miR-671-5p and LINC00943 or ELAVL1 was confirmed by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Our data validated that LINC00943 regulated MPP-evoked injury in SK-N-SH cells. LINC00943 regulated miR-671-5p expression by binding to miR-671-5p. Moreover, miR-671-5p was identified as a molecular mediator of LINC00943 in regulating SK-N-SH cell injury induced by MPP. MiR-671-5p targeted and inhibited ELAVL1, and miR-671-5p-mediated inhibition of ELAVL1 impacted MPP-evoked SK-N-SH cell injury. Furthermore, LINC00943 involved the post-transcriptional regulation of ELAVL1 through miR-671-5p competition. Our present study has established a novel mechanism, the LINC00943/miR-671-5p/ELAVL1 ceRNA crosstalk, for the regulation of LINC00943 on MPP toxicity in SK-N-SH cells.
长链非编码 RNA（lncRNA）的竞争内源性 RNA（ceRNA）活性在包括帕金森病在内的病理紊乱中具有深远的影响。在这里，我们专注于长链非编码 RNA00943 介导的 ceRNA 网络，以调节 SK-N-SH 细胞中 MPP 毒性的 LINC00943。将 SK-N-SH 细胞暴露于 1-甲基-4-苯基吡啶（MPP）中。通过实时定量 PCR（RT-qPCR）或 Western blot 定量 LINC00943、miR-671-5p 和 ELAV 样 RNA 结合蛋白 1（ELAVL1）。通过细胞计数试剂盒-8（CCK-8）测定和流式细胞术分别测定细胞活力和凋亡。通过双荧光素酶报告和 RNA 免疫沉淀（RIP）测定证实了 miR-671-5p 与 LINC00943 或 ELAVL1 之间的直接关系。我们的数据验证了 LINC00943 调节 SK-N-SH 细胞中 MPP 诱发的损伤。LINC00943 通过与 miR-671-5p 结合来调节 miR-671-5p 的表达。此外，miR-671-5p 被鉴定为 LINC00943 调节 MPP 诱导的 SK-N-SH 细胞损伤的分子介质。miR-671-5p 靶向并抑制 ELAVL1，miR-671-5p 介导的 ELAVL1 抑制影响 MPP 诱发的 SK-N-SH 细胞损伤。此外，LINC00943 通过 miR-671-5p 竞争参与 ELAVL1 的转录后调节。本研究建立了一种新的机制，即 LINC00943/miR-671-5p/ELAVL1 ceRNA 串扰，用于调节 LINC00943 在 SK-N-SH 细胞中 MPP 毒性。
