Abstract: Objective：To investigate the effect of low glucose（LG） on the epithelialmesenchymal transition (EMT)associated migration and the role of heat shock factor 1 (HSF1) in it. Methods：The hepatocellular carcinoma (HCC) cells were cultured with LG and high glucose (Control) medium, respectively. Cell migration ability was assessed by the Transwell migration assay. Then, the mRNA and protein levels of HSF1 and EMT-associated markers were determined by real-time polymerase chain reaction (RT-PCR) and Western blot, respectively. In addition, the role of HSF1 in LG-induced inhibition of EMT-associated migration was investigated by the RNA interference (shRNA) and HSF1 expression rescue (shRES) strategy. Results：LG inhibited the EMT-like morphological change, cadherin switching and migration of HCC cells. Meanwhile, LG could down-regulate the expressions of HSF1 and N-Cadherin, but up-regulate the expression of E-Cadherin. In addition, in the HSF1knockdown HCC cells, LG but not Control enhanced the migration ability and EMT of HCC cells (P<0.05), increased the expression of N-Cadherin, and decreased the expression of E-Cadherin. shRES could inhibit the migration ability and EMT of HCC cells again. Conclusion：LG can down-regulate the expression of HSF1 and inhibit the migration ability and EMT of HCC cells, which may provide a novel and effective therapeutic strategy for the treatment of HCC.