P-0003 - Effect of MIIP/IIP45 re-expression by gene transfection on the invasion and metastasis of gastric carcinoma cells in vitro
|Date||28 June 2014|
|Event||World GI 2014|
|Topics|| Basic Science
|Citation||Annals of Oncology (2014) 25 (suppl_2): ii14-ii104. 10.1093/annonc/mdu165|
Y. Wang1, Y. Xin2
The gene of migration and invasion inhibitory protein (MIIP) was first found by Song WE in glioma cells in 2003, and located at chromosome 1p36. The loss of MIIP gene induces different types of tumors. There has been no report about the relationship between MIIP and gastric cancer to date. PAK4, as the representative member of the two classes of PAKs, involves in a wide range of biological behaviours, including inhibition of apoptosis and cell adhesion, promotion of cell migration and tumorigenesis. In this study we investigated the effect of re-expression of MIIP by gene transfection on migration and invasion of human gastric cancer cell lines BGC823 and MKN45, and evaluated the relationship between MIIP and PAK4 pathway.
MIIP mRNA and protein expression in human normal gastric epithelial cell line GES-1 and three gastric cancer cell lines BGC823, MKN45 and SGC7901 were evaluated using qRT-PCR and western blot method. Lentiviral plasmid of MIIP gene was transfected into gastric cancer cell line BGC-823 and MKN-45 to create MIIP gene re-expression in gastric cancer cells. MTT and transwell method were used to evaluate the influence of MIIP on proliferation and migration as well as invasion of gastric cancer cells. Semi quantitative Western blot was used to assess the changes of molecules in the downstream pathways.
GES-1 cells expressed high level of MIIP mRNA and protein, while gastric cancer cell lines BGC823 and MKN45 expressed very low level of MIIP. Lentiviral plasmids were successfully transfected into gastric cancer cell line BGC-823 and MKN-45 and two kinds of gastric cancer cells (named BGC823-MIIP and MKN45-MIIP) with MIIP re-expression were created. Compared with control BGC823 and MKN45 tumor cells, the migration and invasion of BGC823 and MKN45 tumor cells with MIIP re-expressed were significantly inhibited in vitro. Re-expression of MIIP down-regulated the expressions of PAK4, PI3K, p-Limk-1 and p-confflin proteins in BGC823 and MKN45 tumor cells. Meanwhile, MIIP re-expression leads to tubulin deacetylation.
We reported for the first time that MIIP is significantly down-regulated in human gastric cancer cell lines BGC823 and MKN45 compared to human normal gastric epithelial cell line GES-1. Re-expression of MIIP inhibited the migration and invasion of gastric cancer cells in vitro. MIIP re-expression in gastric cancer cells lead to tubulin deacetylation. Meanwhile, MIIP re-expression down regulated PAK4, PI3K, p-Limk-1 and p-confflin protein expression. The further study is needed to reveal the role and relevant mechanism of MIIP in gastric carcinogenesis, invasion and metastasis of gastric cancer.