Objective: To explore the biological function of LINC00174 in multiple myeloma (MM).
Methods: Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of LINC00174 and miR-150 in peripheral blood of MM patients and MM cell lines. EdU staining and flow cytometry were used to detect the effects of LINC00174 and miR-150 on the proliferation and apoptosis of MM cells. Western blot was used to detect the expressions of proliferation marker nuclear-related antigen Ki67, apoptosis-related protein cleaved caspase-3 and transcription factor forkhead box protein P1 (FOXP1). Bioinformatics and dual-luciferase reporter assay were used to verify the targeting relationship between LINC00174 and miR-150 and the targeting relationship between miR-150 and FOXP1.
Results: The level of LINC00174 was significantly increased in peripheral blood of MM patients and MM cell lines (P <0.05). Compared with NC-siRNA group, the expression of LINC00174 was significantly reduced in LINC00174-siRNA group, the proliferation of U266 cells was reduced, the apoptosis rate was significantly increased, the level of Ki67 protein was reduced, and the level of cleaved caspase-3 protein was increased (all P <0.05). LINC00174 targeted regulation of the expression of miR-150. Compared with LINC00174-siRNA+NC inhibitor group, the expression of miR-150 in U266 cells in LINC00174-siRNA+miR-150 inhibitor group was significantly reduced, the cell proliferation was enhanced, the apoptosis rate was reduced, the level of Ki67 protein was increased, and the level of cleaved caspase-3 was decreased (all P <0.05). FOXP1 is the target gene of miR-150. Compared with NC mimic group, the expression of FOXP1 protein in miR-150 mimic group was significantly reduced, the cell proliferation was reduced, the apoptosis rate was significantly increased, Ki67 protein level was decreased, and the level of cleaved caspase-3 was increased. Compared with miR-150 mimic + vector group, the expression of FOXP1 protein in miR-150 mimic + pcDNA-FOXP1 group was significantly increased, the cell proliferation was enhanced, the apoptosis rate was reduced, the level of Ki67 protein was increased, and the level of cleaved caspase-3 was decreased (all P <0.05).
Conclusion: LINC00174 promotes the proliferation of MM cells and inhibits cell apoptosis by regulating the miR-150/ FOXP1 axis.
题目: LINC00174对多发性骨髓瘤恶性增殖的影响及其相关作用机制研究.
目的: 探讨LINC00174在多发性骨髓瘤（MM）中的生物学功能。.
方法: 实时荧光定量聚合酶链反应（RT-qPCR）检测MM患者外周血和MM细胞系中LINC00174、miR-150的表达。EdU染色和流式细胞术检测LINC00174和miR-150对MM细胞U266增殖和凋亡的影响。Western blot检测细胞中增殖标志物细胞核相关抗原Ki67、凋亡相关蛋白cleaved caspase-3以及叉头框转录因子P1（FOXP1）蛋白的表达。利用生物信息学、双荧光素酶报告基因实验来验证LINC00174和miR-150之间的靶向关系以及miR-150和 FOXP1之间的靶向关系。.
结果: LINC00174水平在MM患者外周血和MM细胞系中显著升高（P <0.05）。与NC-siRNA组比较，LINC00174-siRNA组中LINC00174的表达显著降低，U266细胞增殖减少，细胞凋亡率显著升高，Ki67蛋白水平降低，cleaved caspase-3蛋白水平升高（均P <0.05）。LINC00174靶向调控miR-150的表达：与LINC00174-siRNA+NC inhibitor组比较，LINC00174-siRNA+ miR-150 inhibitor组U266细胞中miR-150的表达显著降低，细胞增殖能力增强，细胞凋亡率降低，Ki67蛋白水平升高，cleaved caspase-3水平降低（均P <0.05）。 FOXP1是miR-150的靶基因：与NC mimic组比较，miR-150 mimic组细胞中的FOXP1蛋白表达显著降低，细胞增殖减少，细胞凋亡率显著升高，Ki67蛋白水平降低，cleaved caspase-3水平升高；与miR-150 mimic + Vector组比较，miR-150 mimic + pcDNA-FOXP1组细胞中FOXP1蛋白的表达显著升高，细胞增殖能力增强，细胞凋亡率降低，Ki67蛋白水平升高，cleaved caspase-3水平降低（均P <0.05）。.
结论: LINC00174通过调控miR-150/ FOXP1轴促进多发性骨髓瘤细胞增殖，抑制细胞凋亡。.
Keywords: FOXP1; LINC00174; miR-150; multiple myeloma; proliferation.