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The aim of this study was to investigate the effect of short hairpin RNA(shRNA) targeting autotaxin (ATX) on the migratory and invasive capability ofAGS human gastric cancer cells and the growth of xenografts in nude mice. pSUPER-ATXand pSUPER-mock (non-specific), which were constructed corresponding to the ATX-shRNAand negative control mock-shRNA synthesized based on gene sequence, were transfectedwith blank plasmid pSUPER-control into AGS human gastric cancer cells using Lipofectamine.At 24, 48 and 72 h post-transfection, the cells were harvested and analyzed. Theendogenous ATX mRNA and protein of the different groups of AGS cells were detectedby RT-PCR and western blot assays. Cell proliferation was measured by MTT assay.In vitro Transwell and Matrigel assays were used to detect the cell migratoryand invasive capabilities. A tumor xenograft model was generated by subcutaneousinjection of AGS cells into the dorsum of nude mice. The growth of xenograft tumorswas monitored and measured; changes in tumor morphology and the organs of micewere observed by H&amp;E staining. The expression of ATX, MMP-2 and MMP-9 proteinin xenograft tumor tissues was detected by immunohistochemistry and western blotting.In vitro, both the mRNA and protein levels of ATX in the pSUPER-ATX group weresignificantly downregulated (P&lt;0.01), and the cell proliferative, migratoryand invasive capabilities were also significantly decreased. In vivo, no obviousdamage to the organs was found. pSUPER-ATX significantly suppressed the tumorvolume and weight from the 7th week after cell transplantation, compared to thepSUPER-mock, pSUPER-control and WT groups; the inhibition rates were approximately50% (P&lt;0.05). However, no significant differences in these parameters werefound among the WT, pSUPER-mock and pSUPER-control groups. Furthermore, ATX, MMP-2and MMP-9 were expressed at significantly lower levels in the pSUPER-ATX groupcompared to the levels in the other three groups (P&lt;0.05), and a significantcorrelation between ATX and MMP-2 expression was found (r=0.869, P&lt;0.01). Thespecific shRNA targeting ATX downregulated the endogenous ATX of AGS human gastriccancer cells, and inhibited AGS cell proliferative, migratory and invasive potentials.Moreover, shRNA targeting ATX inhibited the growth of human gastric cancer xenograftsin nude mice.

oncology reports

Short hairpin RNA targeting autotaxin reduces human gastric carcinoma AGS cell proliferative, migratory and invasive capabilities in vitro and causes tumor regression in vivo