Effect of hepatocyte cell adhesion molecule regulating protein kinase C ε on proliferation and migration of renal carcinoma 786-O cells

Abstract:

Objective：To investigate the effect of re-expression of hepatocyte cell adhesion molecule（HepaCAM） on the proliferation and migration ability of renal carcinoma 786-O cells. Methods：Protein kinase C ε（PKCε） membrane protein level，PKCε cytosolic protein level and total protein level were determined by Western blot after 786-O cell being infected with Ad-GFP-HepaCAM and Ad-GFP. Matrix metalloproteinase（MMP）-9 and cyclinD1 protein levels were also detected by Western blot after being treated with Ad-GFP-HepaCAM and εV1-2（10 μmol/L，24 h）. The proliferation ability and migration ability of 786-O cells were analyzed by colony formation assay and wound healing assay，respectively. Results：When compared with those of control group，PKCε membrane protein level was decreased（F=82.724，P=0.000；P=0.000） and cytosolic protein level was increased（F=537.006，P=0.000；P=0.000），but PKCε total protein level was not changed（F=0.548，P=0.605；P=0.805） after being infected with Ad-GFP-HepaCAM. After expo－sure to Ad-GFP-HepaCAM and εV1-2，MMP-9 protein level was down-regulated（F=33.109，P=0.001，P=0.000；F=96.868，P=0.000，P=0.000） and cyclinD1 protein level was also down-regulated（F=562.626，P=0.000，P=0.000；F=159.433，P=0.000，P=0.000） when compared with those of control group. The colony formation rate was decreased（F=259.918，P=0.000；P=0.000） and migration ratio was also reduced（F=550.169，P=0.000；P=0.000） in Ad-GFP-HepaCAM group compared with those of εV1-2 group. Conclu－sion：HepaCAM could inhibit 786-O cell proliferation and mi－gration via blocking PKCε translocating from cytoplasm to membrane and inactivating PKCε partially.