The aim of this study was to evaluate the effects and molecular mechanisms of 
everolimus on Panc-1 human pancreatic cancer cells. Panc-1 human pancreatic 
cancer cells were treated with everolimus (10 μg/ml) at selected time points (6, 
12 and 24 h). Cell proliferation and apoptosis were evaluated by MTT and flow 
cytometric analyses. The glycolytic activity was determined by measuring the 
activity of the key enzyme lactate dehydrogenase (LDH) and lactate production. 
The activity of mammalian target of rapamycin (mTOR) signaling was measured by 
western blotting. The expression of genes, including hexokinase 2 (HK2) and 
microRNA-143 (miR-143), was evaluated by real-time polymerase chain reaction 
(PCR). The administration of everolimus time-dependently inhibited proliferation 
and glycolysis and induced apoptosis in the Panc-1 human pancreatic cancer 
cells. As the time of treatment with everolimus increased, the mTOR signaling 
activity decreased, indicated by lower phosphorylation levels of S6 kinase; 
however, the phosphorylation levels of mTOR barely changed. Moreover, our data 
showed an everolimus-induced increase in miR-143 and decrease in HK2 in Panc-1 
cells in a time-dependent manner. In conclusion, the current study indicates a 
novel role of everolimus in its antitumor effect as an inhibitor of glycolysis 
in Panc-1 human pancreatic cancer cells. Furthermore, our data highlights the 
significance of exploring the mechanisms of everolimus and miR-143 in malignant 
tumors.