长链非编码 RNA NEAT1/miR-29b-3p/Sp1 形成正反馈回路，调节人多发性骨髓瘤细胞对硼替佐米的耐药性。

Lnc NEAT1/miR-29b-3p/Sp1 form a positive feedback loop and modulate bortezomib resistance in human multiple myeloma cells.

机构信息

Department of Hematology, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chinese Academy of Sciences Sichuan Translational Medicine Research Hospital, Chengdu, 610072, Sichuan, China.

Department of Hematology, Dongli Medical District of Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chinese Academy of Sciences Sichuan Translational Medicine Research Hospital, Chengdu, 610051, Sichuan, China.

出版信息

Eur J Pharmacol. 2021 Jan 15;891:173752. doi: 10.1016/j.ejphar.2020.173752. Epub 2020 Nov 28.

DOI:10.1016/j.ejphar.2020.173752

PMID:33253679

Abstract

The overall survival of multiple myeloma (MM) patients significantly improved with the use of proteasome inhibitor such as bortezomib. However, resistance to sorafenib limits its use. Bortezomib-resistant MM cells were generated and their bortezomib-resistant properties were confirmed by cell viability and apoptosis assays. To explore functions and underlying mechanisms of long non-coding RNA nuclear paraspeckle assembly transcript 1 (NEAT1) on bortezomib resistance in MM, MTT assays, ﬂow cytometry analyses, dual luciferase report gene assays, RNA pulldown assays and chromatin immunoprecipitation assays were carried out. NEAT1 and specific protein 1 (Sp1) was upregulated while miR-29b-3p was down regulated in bortezomib-resistant MM cells. NEAT1 promoted Sp1 expression by sponging miR-29b-3p and then enhanced the tolerance of MM cells to bortezomib. Sp1 targeted to NEAT1 promoter region promoting NEAT1 transcription and formed a positive feedback loop. NEAT1 and Sp1 levels were higher and miR-29b-3p was levels were lower in bortezomib-resistant MM patients. NEAT1/miR-29b-3p/Sp1 feedback loop enhanced the tolerance of MM cells to bortezomib. These results indicate potentially valuable targets for overcoming bortezomib resistance for MM.

摘要

多发性骨髓瘤（MM）患者的整体存活率因使用蛋白酶体抑制剂（如硼替佐米）而显著提高。然而，索拉非尼的耐药性限制了其应用。本研究通过细胞活力和凋亡实验生成了硼替佐米耐药的 MM 细胞，并证实了其硼替佐米耐药特性。为了探索长链非编码 RNA 核斑浆组装转录本 1（NEAT1）在 MM 硼替佐米耐药中的作用和潜在机制，进行了 MTT 检测、流式细胞术分析、双荧光素酶报告基因检测、RNA 下拉实验和染色质免疫沉淀实验。结果显示，在硼替佐米耐药的 MM 细胞中，NEAT1 和特异性蛋白 1（Sp1）上调，而 miR-29b-3p 下调。NEAT1 通过海绵吸附 miR-29b-3p 促进 Sp1 表达，从而增强 MM 细胞对硼替佐米的耐受性。Sp1 靶向 NEAT1 启动子区域促进 NEAT1 转录，并形成正反馈环。硼替佐米耐药 MM 患者中 NEAT1 和 Sp1 水平升高，miR-29b-3p 水平降低。NEAT1/miR-29b-3p/Sp1 反馈环增强了 MM 细胞对硼替佐米的耐受性。这些结果表明，针对该反馈环可能为克服 MM 硼替佐米耐药提供有价值的靶点。