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The epithelial membrane proteins (EMP1-3), which belong to the family of peripheral myelin proteins 22-kDa (PMP22), are involved in epithelial differentiation. EMP2 was found to be a downstream target gene of the tumor suppressor gene HOPX, a homeobox-containing gene. Additionally, a dysregulation of EMP2 has been observed in various cancers, but the function of EMP2 in human lung cancer has not yet been clarified.
In this study, a real-time RT-PCR, Western blot and cytoblock analysis were performed to analyze the expression of EMP2. Gain-of-function was achieved by stable transfection with an EMP2 expression vector and loss-of-function by siRNA knockdown. Stable transfection led to overexpression of EMP2 at both mRNA and protein levels in the transfected cell lines H1299 and H2170.
Functional assays including proliferation, colony formation, migration and invasion assays as well as cell cycle analyzes were performed after stable transfection and it was found that the ectopic EMP2 expression resulted in a reduced cell proliferation, migration and invasion as well as a G1 cell cycle arrest. After the EMP2 gene was silenced by the siRNA knockdown, inhibition of the cell invasive property was observed. These phenomena were accompanied by reduced AKT, mTor and p38 activities.
Taken together, the data suggest that the epithelial membrane protein 2 (EMP2) is a tumor suppressor and exerts its tumor suppressive function by inhibiting AKT and MAPK signaling pathways in human lung cancer cells.