Nedd4L modulates the transcription of metalloproteinasea1 and a13 genes to increase the invasive activity of gallbladder cancer

The aim of this study was to examine whether Nedd4L (neural precursor cell expressed, developmentally down-regulated 4-like) participated in gallbladder carcinogenesis. We first immunohistochemically examined the expression of Nedd4L in various gallbladder tissue specimens. Weak immunoreactivity to Nedd4L-specific antibody was observed in normal or dysplastic epithelial cells. Cancer cells in non-invasive regions exhibited little immunoreactivity, whereas strong immunostaining was found in cytoplasm of many invasive cancers, especially at cancer invasive front with desmoplastic reaction. Notably, siRNA-mediated silencing of the Nedd4L gene significantly decreased the Matrigel-invasion activity and collagen invasion activity of cultured gallbladder cancer cells, without affecting the cell growth. The subtractive mRNA hybridization followed by RT-PCR and immunoblotting revealed that down-regulation of Nedd4L significantly decreased the expression of collagenases, matrix metalloproteinase (MMP)-1 and -13, in gallbladder cancer cells. Finally, immunohistochemical staining showed that many Nedd4L-expressing invasive gallbladder cancer cells co-expressed MMP-1 and MMP-13. These results indicated that over-expression of Nedd4L might lead to gallbladder cancer invasion by regulating the transcription of the MMP-1 and MMP-13 genes.