Hepatitis C virus (HCV) core protein is essential for virus particle formation. Using HCV core-expressing and non-expressing Huh7 cell lines, Uc39-6 and Uc321, respectively, we performed comparative proteomic studies of proteins in the 0.5% Triton X-100-insoluble fractions of cells, and found that core-expressing Uc39-6 cells had much lower vimentin content than Uc321 cells. In experiments using vimentin-overexpressing and vimentin-knocked-down cells, we demonstrated that core protein levels were affected by cellular vimentin content. When vimentin expression was knocked-down, there was no difference in mRNA level of core protein; but proteasome-dependent degradation of the core protein was strongly reduced. These findings suggest that the turnover rate of core protein is regulated by cellular vimentin content. HCV production was also affected by cellular vimentin content. Our findings together suggest that modulation of hepatic vimentin expression might enable the control of HCV production.