Sphingosine Kinase-1 Regulates LoVo Cell Invasion and Migration via Activation of ERK1/2 Pathway

Abstract

Objective To investigate the effect of Sphk1 on colon cancer cell invasion and migration. Methods Human colon cancer LoVo cells were divided into three group: LoVo cells were treated using 100 nM Phorbol 12-myristate 13-acetate(PMA) as the Sphk1 activation group,50 μM N,N-dimethyl-D-erythro-sphingosine(DMS) as suppression group,and 0.9%NaCl as control group.Cell invasiveness and migration were detected by Transwell boyden chamber model.Sphk1,ERK1/2,p-ERK1/2 protein expressions were detected by Western blot,MMP-2,MMP-9 and uPA protein levels in the culture medium were detected by enzyme-linked immunosorbent assay(ELISA),and MMP-2,MMP-9 and uPA mRNA expressions in LoVo cells were detected by semi-quantitative reverse transcription-polymerase chain reaction.Results The Sphk1 activator induced the expression of Sphk1 and obviously enhanced LoVo cell invasion and migration capacity,accompanied with the up-regulating of ERK1/2,p-ERK1/2 protein expressions: moreover the protein in culture medium and the mRNA in cells levels of MMP-2,MMP-9 and uPA were elevated.On the contrary,the inhibitor obviously suppressed the protein expression of Sphk1 and cell invaseness and migration,associated with the suppressing of ERK1/2,p-ERK1/2 protein expressions;furthermore,the protein and the mRNA levels of MMP-2,MMP-9 and uPA were down-regulated.Conclusion Sphk1 was able to promote the invasion and migration of LoVo cells,through the activation of ERK1/2 signaling pathway,MMP-2,MMP-9 and uPA mRNA expression were up-regulated.