miR-21对人喉鳞癌细胞Hep2迁移侵袭能力的影响

Effect of miR-21 on migration and invasion ability in human laryngeal squamous carcinoma cell Hep2

目的探讨微小RNA-21(miR-21)对人喉鳞癌细胞Hep2迁移、侵袭能力的影响。方法用脂质体LipofectamineTM2000将miR-21抑制剂和miR-21NC转入人喉鳞癌细胞Hep2中,48h后,运用四甲基偶氮唑蓝(MTT)比色法检测细胞活力,划痕实验检测细胞迁移能力,Transwell小室法检测细胞侵袭能力,蛋白质印迹法检测人第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)/磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)的激活情况,以及基质金属蛋白酶(MMP)2、MMP9与伴有kazal域的富含半胱氨酸的逆转诱导蛋白(RECK)的表达。结果与miR-21NC比较,miR-21抑制剂能明显降低Hep2细胞活力[(0.688±0.043)vs.(0.375±0.012)],抑制细胞迁移能力[(6.57±0.02)μmvs.(20.49±2.18)μm]及细胞侵袭能力[(100.7±10.2)vs.(46.8±4.3)],差异均有统计学意义(P<0.01);同时miR-21抑制剂能明显下调PI3K、MMP2及MMP9的表达(P<0.01),降低Akt的磷酸化水平(P<0.01),并上调PTEN及RECK的表达(P<0.01)。结论 miR-21抑制剂能明显抑制人喉鳞癌细胞Hep2的迁移、侵袭能力,可能与PTEN/PI3K/Akt信号通路有关。

Objective To explore the effect of microRNA-21（miR-21）on the migration and invasion ability in human laryngeal squamous carcinoma cell Hep2.Methods The MTT method was used to detect the viability of Hep2 cells at 48 hafter miR-21 inhibitor and miR-21 NC transferring into Hep2 cells by LipofectamineTM2000.The cell migration ability was detected by using the scratch test.The cell invasion ability was detected by using the Transwell method.The activation of phosphatase and tensin homologue deleted on chromosome 10（PTEN）/phosphatidylinositol 3kinase（PI3K）/protein kinase B（Akt）signal pathway and the expression of matrix metalloproteinase 2（MMP2）,MMP9,reversion inducing cysteine rich protein with kazal motif（RECK）was detected by using the Western blotting.Results Compared with miR-21 NC,miR-21 inhibitor could significantly reduce the Hep2 cell viability[（0.688±0.043）vs.（0.375±0.012）],inhibited the migration ability[（6.57±0.02）μmvs.（20.49±2.18）μm]and invasion ability[（100.7±10.2）vs.（46.8±4.3）],and the differences were statistically significant（P〈0.01）,meanwhile miR-21 inhibitor could down-regulate the expression of PI3 K,MMP2and MMP9（P〈0.01）,and reduced the phosphorylation level of Akt（P〈0.01）,up-regulated the expression of PTEN and RECK（P〈0.01）.Conclusion miR-21 inhibitor can significantly suppress the migration and invasion ability of Hep2,which may be related with the PTEN/PI3K/Akt signal pathway.