Dana P, Kariya R, Vaeteewoottacharn K et al.
Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Oncology research. Dec 2016.
CD147 is a trans-membrane protein that can induce the expression and activity of matrix metalloproteinases (MMP). Expression of CD147 has been shown to potentiate cell migration, invasion, and metastasis of cancer. In this study, the critical role of CD147 in metastasis was elucidated using CD147-overexpressing cholangiocarcinoma (CCA) cells, in vitro and in vivo. The demonstrated molecular mechanism herein, supported that metastasis increased in CD147-overexpressing cells. Five CD147-overexpressing clones (Ex-CD147) were established from a low CD147 expressing CCA cell line, KKU-055, using lentivirus containing pReceiver-Lenti-CD147. The metastatic capability was determined using the tail-vein injection mouse model and an in vitro 3D-invasion assay. Liver colonization was assessed using anti-HLA class I-immunohistochemistry. Adhesion abilities, cytoskeletal arrangements, MMP activities, the expressions of adhesion molecules and epithelial-mesenchymal transitional markers were analyzed. All Ex-CD147 clones exhibited high CD147 expression and high liver-colonization in the tailvein injected mouse model, whereas parental cells lacked this ability. Ex-CD147 clones exhibited metastatic phenotypes, i.e., an increase of F-actin rearrangement, cell invasion and a decrease of cell adhesion. The molecular mechanisms were shown to be via the induction of MMP-2 activity and enhancement of epithelial-mesenchymal transitions. An increase of mesenchymal markers, namely, slug, vimentin and N-cadherin, and a decrease of epithelial markers; E-cadherin and claudin-1, together with the suppression of the adhesion molecule, ICAM-1, were observed in the Ex-CD147 clones. Moreover, suppression of CD147 expression using siCD147 in two CCA cell lines with high CD147 expression significantly decreased cell migration and invasion of these CCA cells. These findings emphasize the essential role of CD147 in CCA metastasis and suggest CD147 as a promising target for effective treatment of CCA.