Abstract: Objective:To explorer the effects of gap junction protein connexin32on motility and invasiveness of hepato-cellular carcinoma cells. Methods:The stable subclone Li-7 Tet-off Cx32in which connexin 32expression could be controlled by doxycyclin was established in hepatocellualr carcinoma Li-7 cells by retrovirus infection. Cell motility and invasive ability was analyzed by in vitro transwell motility and invasion assay. Results:A subclone originated from Li- 7 cells was established, in which both connexin32cDNA and regulatory element Tet-off were stably integrated. Western-blot results showed that exogeneous connexin 32was expressed in the cytoplasm, mainly in Golgi apparatus, and the abnormal accumulation of connexin32protein in the cytoplasm could significantly promote motility and invasiveness of Li- 7 cells. Conclusion:Cytoplasmic accumulation of connexin32protein can promote tumor progression in Li- 7 hepatocellular carcinoma cells.