微小RNA-29b通过调控信号转导及转录激活因子3信号通路对多发性骨髓瘤细胞增殖和侵袭的影响  

Effect of microRNA-29b on proliferation and invasion of multiple myeloma cells by regulating the signal transducer and activator of transcription 3 pathway

作　　者：牛奔 侯丽敏[1] 周伟[1] Niu Ben;Hou Limin;Zhou Wei(Department of Hematology,Shaanzi Provincial People's Hospital,Xi'an 710068,Shaanzi Province,China)

机构地区：[1]陕西省人民医院血液内科,西安710068

出　　处：《国际输血及血液学杂志》2023年第2期149-155,共7页International Journal of Blood Transfusion and Hematology

基　　金：陕西省人民医院科技发展孵化基金项目(2021YJY-19);陕西省自然科学基础研究计划项目(2012JQ4024)。

摘　　要：目的研究微小RNA(miRNA)-29b通过调控信号转导及转录激活因子(STAT)3信号通路对多发性骨髓瘤(MM)细胞增殖和侵袭的影响及其机制。方法选择人MM细胞系U266、正常浆细胞为研究对象。将对数生长期的U266接种至6孔板, 并设置模拟物组(n=6, 加入miRNA-29b模拟物), 抑制物组(n=6, 加入miRNA-29b抑制物), 阴性对照(NC)组(n=6, 加入miRNA-29b NC物)和空白组(n=6, 不作处理)。采用实时荧光定量(RT)-PCR检测U266和正常浆细胞中miRNA-29b相对表达水平。采用Transwell试剂盒检测细胞侵袭能力, 3-(4, 5-二甲基噻唑-2)-2, 5-二苯基四氮唑溴盐(MTT)试剂盒检测细胞增殖活力, 蛋白质印迹法检测细胞中STAT3及其磷酸化STAT3(p-STAT3)蛋白相对表达水平。2组间miRNA-29b相对表达水平的比较, 采用独立样本t检验。多组miRNA-29b相对表达水平、增殖率、侵袭细胞数、STAT3、p-STAT3蛋白相对表达水平的总体比较, 采用单因素方差分析;组间两两比较, 采用LSD-t检验。结果①空白组U266中miRNA-29b相对表达水平为(1.00±0.10), 显著低于正常浆细胞的(4.25±0.41), 差异有统计学意义(t=18.987, P<0.001)。②模拟物组U266的miRNA-29b相对表达水平较NC组升高[(3.87±0.21)比(0.93±0.09)], 抑制物组比NC组下降[(0.34±0.06)比(0.93±0.09)], 并且差异均有统计学意义(LSD-t=39.921、7.975, P<0.001、0.001), NC组和空白组比较, 差异无统计学意义[(0.93±0.09)比(1.00±0.10), LSD-t=1.084, P=0.291]。③对于细胞增殖活力, 培养24 h时, 模拟物组、NC组及抑制物组吸光度值比较, 差异无统计学意义(F=3.722, P=0.089)。培养48、72 h时, 模拟物组吸光度值分别为(0.53±0.05)和(0.91±0.07), 较NC组的(0.72±0.06)和(1.23±0.10)显著降低(LSD-t=3.561、3.982, P=0.012、0.007), 抑制物组分别为(0.90±0.08)和(1.66±0.12), 较NC组显著增高(LSD-t=3.370、5.309, P=0.015、0.002)。④模拟物组U266侵袭细胞数较NC组显著减少[(23.6±2.1)个比(154.4±7.2Objective To study the effect of microRNA(miRNA)-29b on the proliferation and invasion of multiple myeloma(MM)cells by regulating the signal transducer and activator of transcription(STAT)3 signaling pathway and its mechanism.Methods The human MM cell line U266 and normal plasma cells were selected as the research objects.U266 in the logarithmic growth phase was inoculated into 6-well plates,and set up mimic group(n=6,adding miRNA-29b mimic),inhibitor group(n=6,adding miRNA-29b inhibitor),negative control(NC)group(n=6,adding miRNA-29b NC object)and blank group(n=6,no treatment).The relative expression levels of miRNA-29b in U266 and normal plasma cells were detected by real-time fluorescent quantitative(RT)-PCR.Transwell test kit was used to detect cell invasion ability,3-(4,5)-dimethylthiahiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)test kit was used to detect cell proliferation activity,and Western blotting was used to detect the relative expression levels of STAT3 and its phosphorylated STAT3(p-STAT3)protein in cells.The comparison of relative expression levels of miRNA-29b between two groups was performed by independent sample t test.The overall comparison of relative expression level of miRNA-29b,cell proliferation activity,number of invasive cells,relative expression levels of STAT3 and p-STAT3 protein among multiple groups were conducted by one-way analysis of variance;pairwise comparisons between groups were conducted by LSD-t test.Results The relative expression level of miRNA-29b in U266 in the blank group was(1.00±0.10),which was lower than that of normal plasma cells(4.25±0.41),and the difference was statistically significant(t=18.987,P<0.001).The relative expression level of miRNA-29b in the mimic group was significantly higher than that of NC group[(3.87±0.21)0.93±0.09)J,the difference was statistically significant(LSD-t=39.921,7.975;P<0.001,0.001),there was no significant difference between NC group and blank group[(0.93±0.09)us(1.00±0.10);LSD-t=1.084,P=0.291].For cell proliferation activ

关 键 词：多发性骨髓瘤 微RNAS STAT3转录因子 细胞增殖 侵袭 

分 类 号：R733.3[医药卫生—肿瘤]