title: Short hairpin RNA targeting autotaxin reduces human gastric carcinoma             AGS cell proliferative, migratory and invasive capabilities in vitro and causes             tumor regression in vivo.
abstract: The aim of this study was to investigate the effect of short hairpin RNA             (shRNA) targeting autotaxin (ATX) on the migratory and invasive capability of             AGS human gastric cancer cells and the growth of xenografts in nude mice. pSUPER-ATX             and pSUPER-mock (non-specific), which were constructed corresponding to the ATX-shRNA             and negative control mock-shRNA synthesized based on gene sequence, were transfected             with 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. The             endogenous ATX mRNA and protein of the different groups of AGS cells were detected             by 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 migratory             and invasive capabilities. A tumor xenograft model was generated by subcutaneous             injection of AGS cells into the dorsum of nude mice. The growth of xenograft tumors             was monitored and measured; changes in tumor morphology and the organs of mice             were observed by H&E staining. The expression of ATX, MMP-2 and MMP-9 protein             in 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 were             significantly downregulated (P<0.01), and the cell proliferative, migratory             and invasive capabilities were also significantly decreased. In vivo, no obvious             damage to the organs was found. pSUPER-ATX significantly suppressed the tumor             volume and weight from the 7th week after cell transplantation, compared to the             pSUPER-mock, pSUPER-control and WT groups; the inhibition rates were approximately             50% (P<0.05). However, no significant differences in these parameters were             found among the WT, pSUPER-mock and pSUPER-control groups. Furthermore, ATX, MMP-2             and MMP-9 were expressed at significantly lower levels in the pSUPER-ATX group             compared to the levels in the other three groups (P<0.05), and a significant             correlation between ATX and MMP-2 expression was found (r=0.869, P<0.01). The             specific shRNA targeting ATX downregulated the endogenous ATX of AGS human gastric             cancer cells, and inhibited AGS cell proliferative, migratory and invasive potentials.             Moreover, shRNA targeting ATX inhibited the growth of human gastric cancer xenografts             in nude mice.
authors.affiliation: Department of General Surgery, The First Affiliated Hospital of Wenzhou Medical School, Wenzhou, Zhejiang 325000, PR China. zhouxun.chen@googlemail.com
authors.forename: Zhouxun
authors.initials: Z
authors.lastname: Chen
authors.forename: Yingpeng
authors.initials: Y
authors.lastname: Huang
authors.forename: Xian
authors.initials: X
authors.lastname: Shen
authors.forename: Jian
authors.initials: J
authors.lastname: Guo
authors.forename: Guanbao
authors.initials: G
authors.lastname: Zhu
authors.forename: Henning
authors.initials: H
authors.lastname: Dralle
authors.forename: Cuong
authors.initials: C
authors.lastname: Hoang-Vu
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D055785:Gene Knockdown Techniques; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D010727:Phosphoric Diester Hydrolases / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D013274:Stomach Neoplasms / Q000378:metabolism* / Q000473:pathology; D047368:Tumor Burden; D000099189:Lysophospholipase D
chemical_list: D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D010727:Phosphoric Diester Hydrolases; D034741:RNA, Small Interfering; D000099189:Lysophospholipase D; C522361:MMP2 protein, human; C579270:MMP9 protein, human