Effect of the down-regulation of Yes-associated protein on the cloning ability and energy metabolism of gastric cancer cells

ObjectiveTo investigate the effects of the down-regulation of Yes-associated protein (YAP) on the cloning ability and energy metabolism of gastric cancer cells.

MethodsThe YAP small interfering RNA (YAP siRNA) and negative small interfering RNA (siRNA-NC) were transfected into the gastric cancer cell line BGC823, respectively, and the cells without transfection was set as the control (con).The levels of mRNA and protein level of YAP in cells were detected using the qRT-PCR and western blotting, respectively.The cell proliferation activity and cloning ability were determined using the MTT and plate cloning assay, respectively.The level of ATP in cells and content of lactic acid in supernatant of culture medium were determined by kit.The levels of related energy metabolism proteins HK2, PKM2 and apoptotic protein cleaved caspase-3 were determined using western blotting.

ResultsThe levels of YAP mRNA and protein in YAP siRNA cells were significantly lower than those in Con cells (P < 0.05), and the differences of the levels of YAP mRNA and protein between siRNA-NC and Con cells were not statistically significant (P>0.05).The cell proliferation and colony formation rate in YAP siRNA cells were significantly lower than those in Con cells (P < 0.05), and the apoptosis rate in YAP siRNA cells was significantly higher than that in Con cells (P < 0.05).The level of ATP in cells, content of lactic acid in supernatant and levels of HK2 and PKM2 protein in YAP siRNA cells were lower than those in Con cells (P < 0.05), and the level of cleaved caspase-3 in YAP siRNA cells was higher than that in Con cells (P < 0.05).

ConclusionsThe down regulation of YAP can reduce the abilities of clone formation and proliferation, induce the caspase-3 mediated apoptosis, interfer the expression of key enzymes HK2 and PKM2, decrease the level of ATP and inhibit the glycolysis in gastric cancer cells.