目的：探索miR-149-3p 对宫颈癌HeLa 细胞增殖、凋亡、侵袭及迁移的影响及其机制。方法: HeLa 细胞随机分为5组：未转染(HeLa)组、mimic-scramble 组、miR-149 mimic 组、FOXP3 过表达(pc-FOXP3)组、共转染(mimic+pc-FOXP3)组，Mimicscramble为miR-149 mimic 的阴性对照随机序列。荧光素酶实验验证miR-149-3p 与FOXP3 的靶向结合关系。实时定量PCR(qRT-PCR)检测HeLa 细胞中miR-149-3p 表达及FOXP3 的mRNA水平，Western blotting 检测FOXP3 的蛋白水平。CCK-8 检测细胞增殖，流式细胞术分析细胞凋亡，Transwell 实验检测细胞侵袭，划痕实验分析细胞迁移。结果: 荧光素酶实验显示miR-149-3p可靶向结合FOXP3。与未转染组相比miR-149 mimic 组miR-149-3p 表达升高、FOXP3 mRNA水平下降(P<0.01)，而且miR-149mimic 组FOXP3 蛋白水平低于未转染组(P<0.01)、pc-FOXP3 组FOXP3 蛋白水平高于未转染组(P<0.01)，与pc-FOXP3 组相比mimic+pc-FOXP3 组FOXP3 蛋白水平降低(P<0.01)。miR-149 mimic 组细HeLa 胞增殖倍数低于未转染组(P<0.01)，pc-FOXP3 组细胞增殖倍数高于未转染组(P<0.01)，与pc-FOXP3 组相比mimic+pc-FOXP3 组细胞增殖倍数下降(P<0.01)；miR-149 mimic 组HeLa 细胞凋亡率高于未转染组(P<0.01)，pc-FOXP3 组细胞凋亡率低于未转染组(P<0.01)，与pc-FOXP3 组相比mimic+pc-FOXP3组细胞凋亡率上升(P<0.01)；miR-149 mimic 组每个视野下的侵袭细胞数和划痕愈合率低于未转染组(P<0.01)，pc-FOXP3 组每个视野下的侵袭细胞数和划痕愈合率高于未转染组(P<0.01)，与pc-FOXP3 组相比mimic+pc-FOXP3 组每个视野下的侵袭细胞数和划痕愈合率下降(P<0.01)。结论：miR-149-3p 通过靶向FOXP3 抑制宫颈癌HeLa细胞增殖、侵袭和迁移，同时促进细胞凋亡。

Objective: To explore the effects of miR-149-3p on the proliferation, apoptosis, invasion and migration of cervical cancer HeLa cells and the possible mechanisms. Methods: HeLa cells were randomly divided into five groups, including untransfected (HeLa)group, mimic-scramble group (the negative control of miR-149 mimic), miR-149 mimic group, FOXP3 over-expression (pc-FOXP3)group, and co-transfection (mimic+pc-FOXP3) group. The targeted relationship of miR-149-3p and FOXP3 was verified by luciferase assay. The expressions of miR-149-3p and FOXP3 mRNA were tested by quantitative real-time reverse transcription PCR (qRT-PCR).The protein levels of FOXP3 were measured by Western blotting. The proliferation was detected by CCK-8; the apoptosis was tested by flow cytometry, the cell invasion was measured by transwell invasion assay and cell migration was detected by scratch assay. Results:The luciferase assay showed that miR-149-3p could target combine with FOXP3. Compared with untransfected group, the expression of miR-149-3p was increased while mRNA level of FOXP3 was decreased in miR-149 mimic group (all P<0.01). Moreover, the protein level of FOXP3 in miR-149 mimic group was lower than that in untransfected group (P<0.01), while the protein level of FOXP3 in pc-FOXP3 group was higher than that in untransfected group (P<0.01); Compared with pc-FOXP3 group, the protein levels of FOXP3 in mimic+pc-FOXP3 group were reduced (P<0.01). The proliferation in miR-149 mimic group was lower than that in untransfected group (P<0.01), while the proliferation in pc-FOXP3 was higher than that in untransfected group (P<0.01); compared with pc-FOXP3 group,the proliferation in mimic+pc-FOXP3 group was decreased (P<0.01). The apoptosis rate of HeLa cells in miR-149 mimic group was higher than that in untransfected group (P<0.01), while the apoptosis rate in pc-FOXP3 was lower than that in untransfected group (P<0.01); compared with pc-FOXP3 group, the apoptosis in mimic+pc-FOXP3 group was elevated (P<0.01). The number of invasive cells per field and wound healing rate in miR-149 mimic group was lower than those in untransfeccted group (P<0.01) while the invasive cells and wound healing rate in pc-FOXP3 group was higher than those in untransfeceted group (P<0.01); compared with pc-FOXP3 group, the number of invasive cells per field and wound healing rate in mimic+pc-FOXP3 group was reduced (P<0.01). Conclusion:miR-149-3p inhibits proliferation, invasion and migration and promotes apoptosis of cervical cancer HeLa cells via targeting FOXP3.

河南省卫生厅科技攻关项目(No. 201303105)