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

ObjectiveTo 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. ConclusionSphk1 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.