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Vol. 66 No. 3: Issue 3

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

https://doi.org/10.14715/cmb/2020.66.3.22
Xin Xie
Department of Teaching and Research Center, Liaoning University of Traditional Chinese Medicine, Shenyang, 110000, China
Wenjun Liu
Department of Teaching and Research Center, Liaoning University of Traditional Chinese Medicine, Shenyang, 110000, China
Zhiyuan Duan
Department of Teaching and Research Center, Liaoning University of Traditional Chinese Medicine, Shenyang, 110000, China
Xu Li
Foot and Ankle Surgery, Central Hospital Affiliated To ShenYang Medical College, Shenyang, 110000, China
Lide Zhang
Department of Teaching and Research Center, Liaoning University of Traditional Chinese Medicine, Shenyang, 110000, China
Guanlin Yang
Department of Teaching and Research Center, Liaoning University of Traditional Chinese Medicine, Shenyang, 110000, China

Corresponding Author(s) : Guanlin Yang

guanlinyang1988@outlook.com

Cellular and Molecular Biology, Vol. 66 No. 3: Issue 3

Abstract

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.

Keywords

lncRNA NORAD miR-410-3p Osteosarcoma Cisplatin Drug resistance.
Xie, X., Liu, W., Duan, Z., Li, X., Zhang, L., & Yang, G. (2020). LncRNA NORAD targets miR-410-3p to regulate drug resistance sensitivity of osteosarcoma. Cellular and Molecular Biology, 66(3), 143–148. https://doi.org/10.14715/cmb/2020.66.3.22
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