miR-100对大鼠骨髓间充质干细胞迁移能力的影响

目的:探讨MicroRNA-100(miR-100)对大鼠骨髓间充质干细胞(r BMMSC)迁移能力的影响。方法:分离培养r BMMSC,采用流式细胞术检测r BMMSC表面标志物CD105、CD45、CD34、CD29和CD44的表达。用miR-100类似物或抑制剂转染r BMMSC,应用实时定量PCR检测转染后细胞miR-100表达量。迁移实验测定转染后细胞迁移能力,观察miR-100对细胞迁移能力的影响。ELISA法定量检测培养上清中趋化因子SDF-1的分泌水平。结果:经鉴定所分离的细胞确定为r BMMSC。用miR-100类似物或抑制剂转染r BMMSC后,与对照组相比,转染miR-100类似物的细胞miR-100表达显著上调(P<0.01),转染miR-100抑制剂的细胞miR-100表达显著下调(P<0.01)。在迁移实验中,转染miR-100类似物可显著抑制r BMMSC迁移(P<0.01),而转染miR-100抑制剂可显著增强r BMMSC迁移(P<0.01)。转染miR-100类似物组、转染miR-100抑制剂组的培养上清中,SDF-1浓度与对照组相比未见明显变化(P>0.05)。结论:miR-100可显著抑制r BMMSC的迁移,但与趋化因子SDF-1无明显关系。

Effect of miR-100 on Migration of Rat Bone Marrow Mesenchymal Stem Cells

Objective:To explore the effect of miR-100 on the migration of rat bone marrow mesenchymal stem cells(rBMMSC).Methods:The rBMMSC were isolated by density gradient centrifugation,cell surface epitopes of CD105,CD45,CD34,CD29 and CD44 were analyzed by flow cytometry.The rBMMSC were transfected with miR-100 mimic or inhibitor,then the expression of miR-100 in transfected cells was detected by real-time PCR.Migration test was used to observe the effect of miR-100 on cell migration ability.The secretion level of chemokine SDF-1 in culture supernatant of cells was quantitatively detected by ELISA.Results:The isolated cells were identified as BMMSC.After rBMMSC were transfected with miR-100 mimic or inhibitor,as compared with control group,the expression of miR-100 in rBMMSC significantly increased or decreased respectively.In the migration experiment,the rBMMSC migration was significantly inhibited in the miR-100 mimic group(P<0.01),while the rBMMSC migration was significantly enhanced in the miR-100 inhibitor group(P<0.01).The concentration of SDF-1 in the supernatant of the miR-100 mimic group and the miR-100 inhibitor group did not change significantly compared with the control group(P>0.05).Conclusion:miR-100 can significantly inhibit the migration of rBMMSC,but not significantly correlated with the SDF-1.