LncRNA NORAD targets miR-410-3p to regulate drug resistance sensitivity of osteosarcoma

• Published in: Cellular and Molecular Biology Vol. 66; no. 3; pp. 143 - 148
• Main Authors: Xie, Xin, Liu, Wenjun, Duan, Zhiyuan, Li, Xu, Zhang, Lide, Yang, Guanlin
• Format: Journal Article
• Language: English
• Published: France 05-06-2020
• Subjects: ATP Binding Cassette Transporter, Subfamily B, Member 1 - genetics; ATP Binding Cassette Transporter, Subfamily B, Member 1 - metabolism; Base Sequence; Bone Neoplasms - drug therapy; Bone Neoplasms - genetics; Bone Neoplasms - pathology; Cell Line, Tumor; Cell Proliferation - drug effects; Cisplatin - pharmacology; Cisplatin - therapeutic use; Drug Resistance, Neoplasm - genetics; Gene Expression Regulation, Neoplastic - drug effects; Humans; MicroRNAs - genetics; MicroRNAs - metabolism; NF-kappa B - metabolism; Osteosarcoma - drug therapy; Osteosarcoma - genetics; Osteosarcoma - pathology; RNA, Long Noncoding - genetics; RNA, Long Noncoding - metabolism; Signal Transduction; miR-410-3p; Drug resistance; lncRNA NORAD; Osteosarcoma; Cisplatin
• ISSN: 0145-5680; 1165-158X
• DOI: 10.14715/cmb/2020.66.3.22

The current experiment was performed to investigate the effect of LncRNA NORAD on the sensitivity of miR-410-3p to drug resistance of osteosarcoma cells. The cisplatin-resistant cell line HOS/DDP was induced; si-NC, si-NORAD, miR-NC, miR-410-3p, pcDNA-NC, and pcDNA-NORAD were transfected into HOS/DDP cells, respectively; record as a si-NC group, si-NORAD group, miR-NC group, miR-410-3p group, pcDNA-NC group, pcDNA-NORAD group; si-NORAD was co-transfected into HOS/DDP cells with anti-miR-NC, anti-miR-410-3p, recorded as an anti-miR-NC+si-NORAD group and anti-miR-410-3p+si-NORAD group. Real-time quantitative PCR (RT-qPCR) was used to detect LncRNA NORAD, miR-410-3p and multidrug resistance protein 1 (MRP1) mRNA expression levels; Western blot was used to detect cyclin D1 (CyclinD1), MRP1, phosphorylated (p-p65), phosphorylated IкBα (p-IкBα) protein expression; cell counting kit 8 (CCK-8) was used to detect cell viability; dual luciferase report assay to detect targeting relationship between LncRNA NORAD and miR-410-3p. Compared with osteosarcoma cells HOS, the expression levels of LncRNA NORAD, MRP1 mRNA and protein in osteosarcoma resistant cells HOS/DDP were significantly increased, and miR-410-3p expression levels were significantly reduced (P<0.05). Low expression of NORAD or high expression of miR-410-3p, CyclinD1, MRP1 expression levels were significantly reduced, the cell survival rate was significantly reduced, and the half inhibitory concentration of cisplatin was significantly reduced (P<0.05). LncRNA NORAD targets and regulates miR-410-3p, and low expression of miR-410-3p can reverse the effects of low NORAD expression on HOS/DDP cell proliferation and cisplatin resistance. Low expressions of NORAD, p-p65, p-IкBα protein expression levels were significantly reduced; low expression of miR-410-3p could reverse the inhibitory effect of low NORAD expression on p-p65, p-IкBα protein expression. Inhibition of LncRNA NORAD expression can inhibit the proliferation of osteosarcoma HOS/DDP cells through targeted regulation of miR-410-3p, increasing its sensitivity to cisplatin, and it may be related to the NF-κB signaling pathway.