Effect of Map3k1 gene down regulation on proliferation and migration abilities of B 6 mouse eyelid keratinocytes / 重庆医学

ABSTRACT

Objective To study the effect of targeted inhibition of mitogen-activated protein kinase kinase kinase 1(Map3k1) gene on the cell proliferation and migration abilities of B6 mouse eyelid keratinocytes.Methods An artificial microRNA(amiRNA) interference vector targeting silent Map3k1 gene was constructed in vitro in the test.Lipofectamin 2000 was used to transfect the B6 mouse eyelid keratinocytes(the Ctrl Map3k1 group was transfected with empty vector,while the Map3k1 amiRNA-3 group was transfected with Map3k1 amiRNA-3 interference vector).The Map3k1 mRNA and protein expression levels were respectively de-tected by real-time PCR and Western-blot for determining the interference efficiency.The B6 mouse eyelid keratinocytes prolifera-tion level was detected by MTT.The migration ability of keratinocytes was detected by the scratch experiment.Results After the keratinocytes were transfected with Map3k1 amiRNA interference vector,the levels of Map3k1 mRNA and protein were effectively inhibited,and the interference efficiency was up to 70%(P<0.05).The proliferation level of keratinocytes in the Map3k1 amiRNA-3 group was lower than that in the Ctrl Map3k1 group(P<0.05).The migratory ability of keratinocytes in the Map3k1 amiRNA-3 group was also significantly lower than that in the Ctrl Map3k1 group(P<0.05).Conclusion Targeted inhibition of Map3k1 gene expression in B6 mouse eyelid keratinocytes significantly inhibits cell proliferation and migration,thus influence the cellular bi-ological behaviors.