miR-106b对人肝癌细胞HepG2细胞迁移和侵袭能力的影响

Effects of miR-106b on migration and invasion of HepG2 cell

目的探讨miR-106b对人肝癌细胞HepG2迁移和侵袭能力的影响。方法分别用阴性对照(NC组)、miR-106b模拟物(miR-106b组)和miR-106b抑制物(miR-106b-in组)转染HepG2细胞,实时PCR检测三组HepG2细胞miR-106b的表达,划痕实验、Transwell实验、3D培养实验检测三组HepG2细胞的迁移及侵袭能力的差异。结果实时PCR显示miR-106b组细胞miR-106b相对表达量为(17.83±1.66),miR-106b-in组为(0.32±0.07),差异有统计学意义(P<0.05);NC组细胞24 h及48 h划痕修复率分别为(47±3)%、(71±6)%,miR-106b组分别为(65±5)%、(99±1)%,miR-106b-in组分别为(38±3)%、(56±4)%,组间两两比较差异均有统计学意义(P<0.05);Transwell侵袭实验显示,NC组的穿室细胞数为(58±10)个,miR-106b组为(116±19)个,miR-106b-in组为(38±6)个,组间两两比较差异均有统计学意义(P<0.05);3D培养实验显示,20个200倍视野下,miR-106b组无伪足的球形细胞数为(3±1)个,明显少于NC组的(7±2)个及miR-106b-in组的(11±2)个,差异均有统计学意义(P<0.05)。结论 miR-106b能明显增强HepG2细胞的迁移和侵袭能力。

Objective To investigate the influence of miR-106b on HepG2 cell＇s migration and invasion.Methods HepG2 cells were respectively transfected with negative control （NC group）, miR-106b analogue （miR-106bgroup） and miR-106b inhibitor （miR-106b-in group）. Real-time PCR was used to detect the expression of miR-106b inHepG2 cells. The differences of HepG2 cells＇ migration and invasion in the three groups were detected and compared byWound Healing assay, Transwell assay and 3D spheroid invasion assay. Results Real-time PCR showed that the cells＇miR-106b relative expression of the miR-106b group was （17.83 ± 1.66）, which was significantly higher than （0.32 ±0.07） of the miR-106b-in group （P〈0.05）. The cells＇ 24 h and 48 h scratch repair rates of the NC group were （47±3）%and （71±6）% respectively, which were significantly lower than corresponding （65±5）% and （99±1）% of the miR-106bgroup （P〈0.05）. The Transwell experiment indicated that the migrated cells count in the NC group, the miR-106b groupand the miR-106b-in group were respectively （58±10）, （116±19） and （38±6）, and the differences among the three groupswere statistically significant （P〈0.05） as well. Under 200 magnified visual field, 3D culture experiment expressed that thenumbers of globular cells without pseudopodia in the miR-106b group, miR-106b-in group, NC group were （3±1）, （11±2）,and （7±2）, respectively, and the differences were statistically significant （P〈0.05）. Conclusion MiR-106b can signifi-cantly enhance HepG2 cell＇s migration and invasion.