抑制miR-23a-3p表达对人白血病细胞株HL-60侵袭和迁移的调节作用  

Regulating effect of inhibiting expression of miR-23a-3p on invasion and migration of human leukemia cell line HL-60

作　　者：胡敏[1] 符才波 陈瑜 王娇 陶石[1] HU Min;FU Cai-bo;CHEN Yu;WANG Jiao;TAO Shi(Department of Hematology,the First Affiliated Hospital of Hainan Medical University,Haikou 570102,China)

机构地区：[1]海南医学院附属第一医院血液内科,海口570102

出　　处：《临床与实验病理学杂志》2018年第8期885-888,892,共5页Chinese Journal of Clinical and Experimental Pathology

基　　金：海南省卫生厅项目(1341320.37A2010)

摘　　要：目的探讨抑制miR-23a-3p表达对人白血病细胞株HL-60侵袭和迁移的影响及其分子机制。方法转染miR-23a inhibitor到HL-60细胞抑制miR-23a-3p的表达。HL-60细胞分为3组:HL-60组(阴性对照组)、scramble组(转染对照组)和miR-23a inhibitor组。qRT-PCR技术检测MMP-9和VEGF mRNA水平,Transwell实验检测细胞侵袭,划痕实验分析细胞迁移。Western blot法检测MMP-9和VEGF蛋白表达。结果转染scramble后HL-60细胞miR-23a-3p表达无明显变化。转染miR-23a inhibitor后HL-60细胞miR-23a-3p表达显著下降(P<0.01)。scramble组与阴性对照组每个视野下的侵袭细胞数目差异无统计学意义。miR-23a inhibitor组每个视野下的侵袭细胞数目显著低于阴性对照组(P<0.01)。scramble组与阴性对照组细胞迁移无明显差异。与阴性对照组相比,miR-23a inhibitor组细胞迁移显著降低(P<0.01)。转染scramble不会影响MMP-9和VEGF的表达。miR-23a inhibitor组MMP-9和VEGF的mRNA水平显著低于阴性对照组(P<0.01)。与阴性对照组相比,miR-23a inhibitor组MMP-9和VEGF的蛋白水平显著下降(P<0.001)。结论抑制miR-23a-3p表达可减弱人白血病细胞株HL-60的侵袭和迁移。Purpose The study aims to explore the effect of inhibiting expression of miR-23a-3p on invasion and migration of human leukemia cell line HL-60.Methods The expression of miR-23a-3p was interfered with miR-23a inhibitor.HL-60 cells were divided into three groups:HL-60(negative control)group.scramble(transfection control)group and miR-23a inhibitor group.The mRNA levels of matrix metalloprotein 9(MMP-9)and vascular endothelial growth factor(VEGF)were detected by quantitative real-time reverse transcription PCR(qRT-PCR).Cell invasion was measured by transwell assay.Cell migration was tested by wound healing.The protein levels of MMP-9 and VEGF were detected by Western blot.Results There was no obvious change in expression of miR-23a-3p in HL-60 cells after transfection with scramble.After transfection with miR-23a inhibitor,the expression of miR-23a-3p in HL-60 cells was decreased remarkably(P<0.01).There was no statistical difference in the number of invasive cells per field between the scramble group and negative control group.The number of invasive cells per field in miR-23a inhibitor group was significantly lower than that of negative control group(P<0.01).There was no obvious difference in cell migration between scramble group and negative control group.Compared with negative control group,the migration of miR-23a inhibitor group was markedly reduced(P<0.01).After transfection with scramble,the expression of MMP-9 and VEGF was not affected.The mRNA levels of MMP-9 and VEGF in miR-23a inhibitor group were notably lower than that of negative control group(P<0.01).Compared with negative control group,the protein levels of MMP-9 and VEGF in miR-23a inhibitor group were decreased dramatically(P<0.001).Conclusion Inhibiting expression of miR-23a-3p attenuates cell invasion and migration of human leukemia cell line HL-60.

关 键 词：白血病 侵袭 迁移 miR-23a-3p 

分 类 号：R733.71[医药卫生—肿瘤]