Liu Rong, Wu Min, Xu Guiju, Ju Lu, Xiao Jinhui, Zhong Wei, He Xiao, Yang Yan
Department of Anesthesiology, The 908th Hospital of Chinese PLA Logistical Support Force, No.4, Hudong Road, Yuehu District, Yingtan 335000, Jiangxi, China.
Department of Internal Medicine, Ruijin Hospital of traditional Chinese Medicine, Ruijin, Jiangxi, 342500, China.
Open Life Sci. 2020 Dec 31;15(1):988-999. doi: 10.1515/biol-2020-0108. eCollection 2020.
Regional anesthesia has anti-proliferative and pro-apoptotic effects in various cancers. Therefore, the purpose of this study was to investigate the effects of ropivacaine on the proliferation, migration, invasion, and apoptosis of glioma cells .
Under ropivacaine stimulation conditions, proliferation, apoptosis, migration, and invasion of glioma cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2-tetrazol-3-ium bromide (MTT), flow cytometry, and transwell assays, respectively. Western blot assay was employed to measure the protein expression levels in glioma cells. The expression levels of small nucleolar RNA host gene 16 (SNHG16) and miR-424-5p were assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The interaction relationship between SNHG16 and miR-424-5p was predicted and confirmed using a bioinformatics database and dual-luciferase reporter, RNA immunoprecipitation (RIP) and RNA pull-down assays.
After treatment with ropivacaine, proliferation, migration, and invasion were repressed while apoptosis was enhanced in glioma cells in a dose-depended manner. In addition, ropivacaine impeded SNHG16 expression in glioma cells. Importantly, overexpression of SNHG16 abolished the ropivacaine-induced effects on glioma cells. Analogously, knockdown of miR-424-5p counteracted the function of ropivacaine in glioma cells. We also found that SNHG16 bound to miR-424-5p and negatively regulated miR-424-5p expression in glioma cells. The rescue experiments indicated that ropivacaine might regulate glioma progression by targeting the SNHG16/miR-424-5p axis.
Our findings revealed the anti-tumor effects of ropivacaine in glioma by targeting the SNHG16/miR-424-5p axis. These data might extend the understanding of regulatory mechanisms by which ropivacaine could suppress glioma development.
区域麻醉在多种癌症中具有抗增殖和促凋亡作用。因此，本研究旨在探讨罗哌卡因对胶质瘤细胞增殖、迁移、侵袭和凋亡的影响。
在罗哌卡因刺激条件下，分别通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2-四氮唑溴盐(MTT)、流式细胞术和Transwell实验测定胶质瘤细胞的增殖、凋亡、迁移和侵袭。采用蛋白质免疫印迹法检测胶质瘤细胞中的蛋白表达水平。通过逆转录-定量聚合酶链反应(RT-qPCR)评估小核仁RNA宿主基因16(SNHG16)和miR-424-5p的表达水平。使用生物信息学数据库、双荧光素酶报告基因、RNA免疫沉淀(RIP)和RNA下拉实验预测并证实SNHG16与miR-424-5p之间的相互作用关系。
罗哌卡因处理后，胶质瘤细胞的增殖、迁移和侵袭受到抑制，而凋亡以剂量依赖的方式增强。此外，罗哌卡因抑制胶质瘤细胞中SNHG16的表达。重要的是，SNHG16的过表达消除了罗哌卡因对胶质瘤细胞的诱导作用。类似地，miR-424-5p的敲低抵消了罗哌卡因在胶质瘤细胞中的作用。我们还发现SNHG16与miR-424-5p结合并负调控胶质瘤细胞中miR-424-5p的表达。挽救实验表明，罗哌卡因可能通过靶向SNHG16/miR-424-5p轴调节胶质瘤进展。
我们的研究结果揭示了罗哌卡因通过靶向SNHG16/miR-424-5p轴对胶质瘤具有抗肿瘤作用。这些数据可能会扩展对罗哌卡因抑制胶质瘤发展的调控机制的理解。
