, Volume 28, Issue 2, pp 494-501
Date: 15 Apr 2010

Blockade of proliferation and migration of gastric cancer via targeting CDH17 with an artificial microRNA

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Abstract

Liver-intestine cadherin (CDH17) is a novel member of the cadherin superfamily implicated in gastric cancer progression. To determine the role of CDH17 in the process of gastric cancer invasive growth, in the present study, RNA interference mediated by recombinant lentivirus vectors expressing artificial CDH17 miRNA was applied to induce a long-lasting down-regulation of CDH17 gene expression in BGC823 cells. The expression levels of CDH17, tumor cell motility, migration potential, and proliferation were measured by flow cytometry, real-time RT–PCR, Western blot analysis, immunofluorescence staining, wound healing assay, and MTT assay, respectively. Results show that four recombinant plasmid expression vectors encoding pre-miRNA against CDH17, pcDNA-CDH17-miR-SR1, -SR2, -SR3, and -SR4 were constructed correctly and down-regulated the CDH17 mRNA levels by 5.5, 57, 91, and 98%, respectively, in BGC823 cells which had an overexpression of CDH17. We packaged the recombinant lentiviral vector for CDH17 RNA interference with pcDNA-CDH17-miR-SR4 which had the highest interfering efficiency and succeeded in construction of the stable transfectants. Of note, more than 90% knockdown of CDH17 expression in BGC823 cells was obtained by miRNA technique. The CDH17-miRNA-transfected cells showed significant decrease in cell proliferation, cell motility, and migration in comparison with the control cells. Thus, we proposed that CDH17 may be an oncogene up-regulating invasive features of gastric cancer cells and could be a hopeful target for the control of gastric cancer progression.

Jin Zhang and Qi-Sheng Liu are contributed equally to this work.