MiR-498通过下调FOXM1抑制肺腺癌细胞上皮充质细胞转化

MiR-498 inhibits A549 cells EMT by targeting FOXM1

目的研究miR-498对肺腺癌细胞上皮充质细胞转化(EMT)的影响及相关机制。方法转染miR-498mimic和miR-NC至离体培养的A549细胞中,将细胞分为空白对照组、miR-498组和空质粒组3组,Transw ell法检测细胞迁移侵袭情况,Western blotting检测A549细胞中E-cadherin、fibronectin、vimentin、FOXM1的表达,利用荧光素酶实验验证miR-498的靶基因;再通过转染过表达FOXM1的质粒至A549细胞,将细胞分为miR-498组、miR-498+FOXM1组、miR-498+空质粒组,检测A549细胞的上述生物学指标。结果与空质粒组相比,转染miR-498mimic后,A549细胞中E-cadherin表达增加(P=0.001),fibronectin、vimentin、FOXM1表达减少(P均<0.01),A549细胞的迁移能力减少(P=0.001);荧光素酶实验结果显示,miR-498能够显著降低FOXM1-3'-UTR质粒的荧光素活性(P=0.001);与miR-498+空质粒组相比,转染过表达FOXM1质粒后,A549细胞中E-cadherin表达减少(P=0.002),fibronectin、vimentin表达增加(P均<0.001),A549细胞的迁移能力增加(P=0.001)。结论 miR-498通过下调FOXM1从而抑制A549细胞的EMT。

Objective To investigate the effect of miR-498 on A549 cells epithelial-to-mesenchymal transition （EMT） and its correlated mechanism. Methods After miR-498 was transfected into A549 cells, the cells were divided into control group, empty vector group and miR-498 group; Transwell assay was employed to test the migration ability of A549 cells, Western blotting was used to investigate the expressions of E-cadherin, fibronectin, vimentin and FOXM1 in A549 cells. Luciferase assay was used to confirmed whether FOXM1-3＇-UTR was the target gene of miR-498. Then, the cells were divided into miR-498 group, miR-498 ＋ FOXM1 group, and miR-498 ＋ empty plasmid group, and the above biological indicators of A549 cells were detected again. Results Compared with the empty vector group, the ex- pression of E-cadherin was increased（ P = 0.001 ）, the expression of fibronectin, vimentin and FOXMlwere decreased （all P 〈 0.01 ）, and the migration ablitiy of A549 cells was decreased in the miR-498 group （P = 0. 001 ） ; the Lucifer- ase activity of the FOXM1-3＇-UTR plasmid was significantly suppressed by miR-498 （ P = 0.001 ） ; over-expression of FOXM1 could reverse the effect of miR-498 on A549. Conclusion miR-498 inhibits A549 cells EMT by down-regula- ting FOXM1 expression.