Abstract 391: Short hairpin RNA targeting autotaxin reduces the human gastric carcinoma AGS cell proliferation, migratory and invasive capability in vitro and causes tumor regression in vivo.
Authors: Chen Zhouxun , Huang Yingpeng , Shen Xian , Dralle Henning , Hoang-Vu Cuong , Gimm Oliver 
Published: August 2014

Abstract
To investigate the effect of short hairpin RNA(shRNA) targeting autotaxin on the migratory and invasive capability of human gastric cancer cell AGS and the growth xenografts in nude mice. The pSUPER-ATX and pSUPER-mock (non-specific), which were constructed in corresponding to the ATX-shRNA and negative control mock-shRNA synthesized on basis of gene sequence, were transfected with blank plasmid pSUPER-control into human gastric cancer cell AGS by Lipofectamine. At 24 h, 48 h and 72 h post-transfection, the cells were harvested and analyzed. The endogenous ATX mRNA and protein of different group AGS cells were detected by RT-PCR and western blot assays. The cell proliferation was measured by MTT assay. In vitro Transwell and Matrigel assay were used to detect the cell migratory and invasive capability. Tumor xenograft model was generated by subcutaneous injection of AGS cells into the dorsum of nude mice. The growth of xenograft tumor was monitored and measured; change of the tumor morphology and mice organs were observed by HE staining. The expressions of autotaxin, MMP-2 and MMP-9 protein in xenograft tumor tissue were detected by immunohistochemistry and western blot. in vitro, both the mRNA and protein level of ATX in pSUPER-ATX group were significantly down-regulated (P&lt;0.01), and the cell proliferation, migration and invasive capability were significantly decreased as well. In vivo, no obvious damage of the organs was found. The pSUPER-ATX significantly suppressed the tumor volume and weight since 7th week after cell transplantation, when compared to the pSUPER-mock, pSUPER-control and WT groups, the inhibition rates were about 50%(P&lt;0.05). However, no significant differences of these parameters were found among the WT, pSUPER-mock and pSUPER-control group. Furthermore, autotaxin, MMP-2/MMP-9 were expressed at significantly lower level in pSUPER-ATX group than in other three groups (P&lt;0.05), and a significant correlation between ATX and MMP-2 expression was displayed (r=0.869, P&lt;0.01). The specific shRNA targeting ATX down-regulated the endogenous ATX of human gastric cancer AGS cells, and could inhibite the AGS cell proliferation, migratory and invasive capability. Moreover, the shRNA targeting autotaxin inhibited the growth of human gastric cancer xenograft in nude mice.
Citation Format: Zhouxun Chen, Yingpeng Huang, Xian Shen, Henning Dralle, Cuong Hoang-Vu, Oliver Gimm. Short hairpin RNA targeting autotaxin reduces the human gastric carcinoma AGS cell proliferation, migratory and invasive capability in vitro and causes tumor regression in vivo. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 391. doi:10.1158/1538-7445.AM2013-391
Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.