摘要
目的探讨长链非编码RNA(lncRNA)PITPNA反义RNA 1(PITPNA-AS1)靶向miR-367-3p调控多发性骨髓瘤细胞增殖、迁移和侵袭的分子机制。方法定量实时聚合酶链反应(qRT-PCR)测定多发性骨髓瘤细胞系RPMI-8226、MM1.S、OPM-2中lncRNA PITPNA-AS1和miR-367-3p表达。在RPMI-8226细胞中转染si-NC(si-NC组)、si-lncRNA PITPNA-AS1(si-lncRNA PITPNA-AS1组)、miR-NC(miR-NC组)、miR-367-3p mimics(miR-367-3p组),或同时转染anti-miR-NC与si-lncRNA PITPNA-AS1(anti-miR-NC+si-lncRNA PITPNA-AS1组)、anti-miR-367-3p与si-lncRNA PITPNA-AS1(anti-miR-367-3p+si-lncRNA PITPNA-AS1组),并将未转染的细胞设为对照组(NC组)。采用Western Blot、细胞计数试剂盒8(CCK-8)、细胞克隆形成实验和Transwell实验分别测定RPMI-8226细胞的基质金属蛋白酶(MMP)2、MMP9蛋白表达、细胞活力、克隆形成和迁移、侵袭。双荧光素酶活性检测实验分析lncRNA PITPNA-AS1和miR-367-3p的靶向关系。结果与成骨细胞MC3T3-E1比较,多发性骨髓瘤细胞系RPMI-8226、MM1.S、OPM-2中lncRNA PITPNA-AS1表达量均增加,miR-367-3p的表达量均减少(P<0.05)。干扰lncRNA PITPNA-AS1或miR-367-3p高表达显著降低RPMI-8226细胞的MMP2蛋白、MMP9蛋白的表达量,并降低细胞活力、克隆形式数、迁移和侵袭数量(均P<0.05)。lncRNA PITPNA-AS1靶向调控miR-367-3p的表达。anti-miR-367-3p可以逆转干扰lncRNA PITPNA-AS1对多发性骨髓瘤细胞RPMI-8226增殖、迁移和侵袭的影响。结论干扰lncRNA PITPNA-AS1通过靶向调控miR-367-3p,从而抑制多发性骨髓瘤细胞的增殖、迁移和侵袭。
Objective To study the molecular mechanism of whether long-chain non-coding RNA(lncRNA)PITPNA antisense RNA 1(PITPNA-AS1)targets miR-367-3p to regulate the proliferation,migration and invasion of multiple myeloma cells.Methods Quantitative real-time polymerase chain reaction(qRT-PCR)was used to determine the expression of lncRNA PITPNA-AS1 and miR-367-3p in multiple myeloma cell lines RPMI-8226,MM1.S,and OPM-2.Transfected si-NC(si-NC group),si-lncRNA PITPNA-AS1(si-lncRNA PITPNA-AS1 group),miR-NC(miR-NC group),miR-367-3p mimics(miR-367-3p group)into RPMI-8226 cells,or transfected anti-miR-NC and si-lncRNA PITPNA-AS1(anti-miR-NC+si-lncRNA PITPNA-AS1 group),anti-miR-367-3p and si-lncRNA PITPNA-AS1(anti-miR-367-3p+si-lncRNA PITPNA-AS1 group)at the same time,and set untransfected cells as control(NC group).Western Blot,cell counting kit 8(CCK-8),cell clone formation experiment and Transwell experiment were employed to determine matrix metalloproteinase(MMP)2,MMP9 protein expression,cell viability,clone formation and migration,and invasion of RPMI-8226 cells.The dual luciferase activity detection experiment analyzed the targeting relationship between lncRNA PITPNA-AS1 and miR-367-3p.Results Compared with osteoblast MC3T3-E1,the expression of lncRNA PITPNA-AS1 in multiple myeloma cell lines RPMI-8226,MM1.S,and OPM-2 all increased,and the expression of miR-367-3p all decreased(P<0.05).Interfering with lncRNA PITPNA-AS1 or high expression of miR-367-3p significantly reduced the expression of MMP2 protein and MMP9 protein in RPMI-8226 cells,and reduced cell viability,number of clones,migration and invasion(P<0.05).lncRNA PITPNA-AS1 targeted and regulated the expression of miR-367-3p.anti-miR-367-3p can reverse the effect of interfering lncRNA PITPNA-AS1 on the proliferation,migration and invasion of multiple myeloma cells RPMI-8226.Conclusion Interfering lncRNA PITPNA-AS1 can inhibit the proliferation,migration and invasion of multiple myeloma cells by targeting miR-367-3p.
作者
饶琦
王丹丹
罗婷
赵佳莉
赵寅
RAO Qi;WANG Dandan;LUO Ting;ZHAO Jiali;ZHAO Yin(Department of Hematology,West China Hospital,Sichuan University,West China School of Nursing,Sichuan University,Chengdu 610041,China)
出处
《西部医学》
2022年第6期791-796,802,共7页
Medical Journal of West China
