MAFG-AS1调控miR-24-3p对急性淋巴细胞白血病细胞Molt4增殖及凋亡的影响

MAFG-AS1 regulates the effect of miR-24-3p on the proliferation and apoptosis of acute lymphoblastic leukemia cells Molt4

目的探讨长链非编码RNA(lncRNA)MAFG-AS1通过调控miR-24-3p对急性淋巴细胞白血病细胞Molt4的增殖和凋亡的影响。方法实时荧光定量PCR(qRT-PCR)分析MAFG-AS1和miR-24-3p在白血病血清、细胞中的表达。将Molt4细胞分为si-NC组、si-MAFG-AS1组、miR-NC组、miR-24-3p mimic组、si-MAFG-AS1+anti-miR-NC组、si-MAFG-AS1+miR-24-3p inhibitor组,分别转染si-NC、si-MAFG-AS1、miR-NC、miR-24-3p mimic、si-MAFG-AS1+anti-miR-NC和si-MAFG-AS1+miR-24-3p inhibitor。采用细胞计数试剂盒8(CCK-8)、流式细胞术、蛋白质印迹实验(Western blot)分别测定细胞增殖、凋亡、Bcl-2相关X蛋白(Bax)和B细胞淋巴瘤/白血病-2(Bcl-2)蛋白表达。双荧光素酶报告实验进行MAFG-AS1和miR-24-3p靶向关系的验证。结果在白血病患者血清、细胞中MAFG-AS1表达升高,miR-24-3p表达降低(P<0.01)。与si-NC组相比,si-MAFG-AS1组Molt4细胞24 h、48 h和72 h的细胞抑制率、凋亡率、Bax蛋白表达量增加,Bcl-2蛋白表达量降低(P<0.01)。MAFG-AS1可以靶向miR-24-3p。与miR-NC组相比,miR-24-3p mimic组Molt4细胞24 h、48 h和72 h的细胞抑制率、凋亡率、Bax蛋白表达量增加,而Bcl-2蛋白表达量降低(P<0.01)。si-MAFG-AS1+miR-24-3p inhibitor组Molt4细胞24 h、48 h和72 h的细胞抑制率、凋亡率、Bax蛋白表达量均低于si-MAFG-AS1+anti-miR-NC组,Bcl-2蛋白表达量高于si-MAFG-AS1+anti-miRNC组(P<0.01)。结论MAFG-AS1在白血病患者中高表达,干扰其表达可通过靶向miR-24-3p抑制急性淋巴细胞白血病细胞Molt4的增殖和促进凋亡。

This study was designed to investigate the effects of long non-coding RNA(lncRNA) MAFG-AS1 on the proliferation and apoptosis of acute lymphoblastic leukemia cells Molt4 by regulating miR-24-3p.The expressions of MAFG-AS1 and miR-24-3p in leukemia serum and cells were analyzed by real-time quantitative PCR(qRT-PCR).Molt4 cells were divided into si-NC group,si-MAFG-AS1 group,miR-NC group,miR-24-3p mimic group,si-MAFG-AS1+anti-miR-NC group,si-MAFG-AS1+miR-24-3p inhibitor group,which were transfected respectively with si-NC,si-MAFG-AS1,miR-NC,miR-24-3p mimic,si-MAFG-AS1+anti-miR-NC,si-MAFG-AS1+miR-24-3p inhibitor.Cell counting Kit-8(CCK-8),flow cytometry,Western blot were applied to determine cell proliferation and apoptosis,as well as the levels of Bcl-2 related X protein(Bax),B cell lymphoma,and B-cell lymphoma/leukemia-2(Bcl-2) proteins;dual luciferase reporter experiment was performed to verify the targeting relationship between MAFG-AS1 and miR-24-3p.Data showed that he expression of MAFG-AS1 in serum and cells of leukemia patients was increased,while the expression of mi R-24-3p decreased(P<0.01).Compared with the si-NC group,the cell inhibition rate,apoptosis rate,and Bax protein expression of Molt4 cells in the si-MAFG-AS1 group at 24 h,48 h,and 72 h increased,while the Bcl-2 protein expression decreased(P<0.01).Dual luciferase reporter experiment indicated that MAFG-AS1 is targeted miR-24-3p.Compared with the miR-NC group,the miR-24-3p mimic group demonstrated higher levels of the cell inhibition rate,apoptosis rate,and Bax protein expression of Molt4 cells at 24 h,48 h,and 72 h,but lower level of Bcl-2 protein expression(P<0.01).The cell inhibition rate,apoptosis rate,and Bax protein expression of Molt4 cells in the si-MAFG-AS1+miR-24-3p inhibitor group at 24 h,48 h and 72 h were lower than those of si-MAFG-AS1+anti-miR-NC group,but Bcl-2 protein expression showed contrary tendency(P<0.01).In conclusion,MAFG-AS1 is highly expressed in leukemia patients,and interfering with its expression can inhibit the proliferation and promote the apoptosis of acute lymphocytic leukemia cells Molt4 by targeting miR-24-3p.