miR-199a-5p介导的lncRNA ANRIL通过调节HIF-1α促进多发性骨髓瘤的恶性增殖

lncRNA ANRIL mediated by miR-199a-5p promotes the malignant proliferation of multiple myeloma by regulating HIF-1α

目的 探讨长链非编码RNA(lncRNA)ANRIL在多发性骨髓瘤(MM)中的生物学功能及其可能的作用机制。方法 采集MM患者与健康志愿者血浆,采用qRT-PCR检测lncRNA ANRIL、微小RNA(miR)-199a-5p和低氧诱导因子-1α(HIF-1α)的相对表达水平。分析lncRNA ANRIL、miR-199a-5p和HIF-1α之间的靶向作用关系,进一步分析lncRNA ANRIL与miR-199a-5p表达及miR-199a-5p和HIF-1α表达的调控关系。敲减lncRNA ANRIL、miR-199a-5p和HIF-1α的mRNA水平后,或者上调lncRNA ANRIL、miR-199a-5p和HIF-1α表达后,采用CCK-8测定U266细胞增殖活力,流式细胞术测定细胞凋亡率,Western blot检测Bax、Bcl-2、caspase3和caspase9蛋白表达。结果 与健康志愿者相比,MM患者血浆lncRNA ANRIL和HIF-1α表达上调。miR-199a-5p在MM患者血浆中表达下调。此外,lncRNA ANRIL可与miR-199a-5p相互作用。HIF-1α被证实是miR-199a-5p的一个靶点。lncRNA ANRIL表达与HIF-1α表达呈正向调控,miR-199a-5p表达与HIF-1α表达呈负向调控。下调lncRNA ANRIL、HIF-1α或过表达miR-199a-5p可显著抑制细胞增殖,促进细胞凋亡。过表达lncRNA ANRIL通过调控miR-199a-5p促进U266细胞增殖。结论 miR-199a-5p介导的lncRNA ANRIL通过调节HIF-1α促进MM的恶性增殖。

Objective To investigate the biological function and possible mechanism of long non-coding RNA(lncRNA) ANRIL in multiple myeloma(MM).Methods Plasma samples were collected from MM patients and healthy volunteers, and the relative expression levels of lncRNA ANRIL,microRNA(miR)-199 a-5 p and hypoxia-inducible factor-1α(HIF-1α) were detected by qRT-PCR.The targeting relationship between lncRNA ANRIL,miR-199 a-5 p and HIF-1α was analyzed, and the regulatory relationship between lncRNA ANRIL and the expression of miR-199 a-5 p, miR-199 a-5 p and HIF-1α was further analyzed.After the mRNA levels of lncRNA ANRIL,miR-199 a-5 p and HIF-1α were down-regulated or the expressions of lncRNA ANRIL,miR-199 a-5 p and HIF-1α were up-regulated, the proliferation activity of U266 cells was determined by CCK-8,and the apoptosis rate of U266 cells was determined by flow cytometry.The protein expressions of Bax, Bcl-2,caspase3 and caspase9 were detected by Western blot.Results Compared with healthy volunteers, the expressions of lncRNA ANRIL and HIF-1α were up-regulated in MM patients, the expression level of miR-199 a-5 p was down-regulated in the plasma of MM patients.In addition, lncRNA ANRIL could interact with miR-199 a-5 p.HIF-1α was confirmed to be a target of miR-199 a-5 p.The expression of lncRNA ANRIL was positively regulated with HIF-1α,and the expression of miR-199 a-5 p was negatively regulated with HIF-1α.Down-regulation of lncRNA ANRIL,HIF-1α or overexpression of miR-199 a-5 p could significantly inhibite cell proliferation and promote cell apoptosis.Overexpression of lncRNA ANRIL could promote the proliferation of U266 cells by regulating miR-199 a-5 p.Conclusion lncRNA ANRIL mediated by miR-199 a-5 p promotes the malignant proliferation of MM by regulating HIF-1α.