BACKGROUND: Overexpression of human tissue inhibi- tors of metalloproteinase-1 (TIMP-1) may play an antitu- mor role in inhibiting hepatocellular carcinoma ( HCC) growth and progression. The aim of the study was to con- struct a recombinant adenovirus vector carrying hTIMP-1 cDNA for liver gene therapy and observe its expression in vitro. METHODS: The full-length cDNA of hTIMP-1 was posi- tively cloned into the adenoviral shuttle vector pAdTrack- CMV, and then cotransformed into competent BJ5183 cells with the adenoviral backbone plasmid pAdEasy-1. Thus, a recombinant adenovirus AdhTIMP-1 containing full-length cDNA of hTIMP-1 was generated by homologous recombi- nation in E. coli. AdhTIMP-1 was then packaged and am- plified in adenoviral packaging cells, or human embryonic kidney 293 cells. The viral titer was checked by green fluo- rescent protein (GFP), and the expression of hTIMP-1 in vitro was detected by the techniques of Western blot and RT-PCR. RESULTS: The recombinant adenovirus vector carrying hTIMP-1 was constructed and confirmed by restriction en- donuclease analysis and DNA sequence analysis. The tran- scription of TIMP-1 mRNA in 293 cells was checked by RT-PCR and TIMP-1 protein could be detected in the cul- ture by Western blot analysis. CONCLUSION: The successful construction of recombi- nant adenoviral vector carrying human TIMP-1 and the ef- fective expression in vitro has laid a foundation for further study of its antitumor function and may pave the way for future application in liver gene therapy. BACKGROUND: Overexpression of human tissue inhibi- tors of metalloproteinase-1 (TIMP-1) may play an antitu- mor role in inhibiting hepatocellular carcinoma (HCC) growth and progression. The aim of the study was to con- struct a recombinant adenovirus vector METHODS: The full-length cDNA of hTIMP-1 was postiped cloned into the adenoviral shuttle vector pAdTrack-CMV, and then cotransformed into competent BJ5183 cells with the adenoviral backbone plasmid pAdEasy-1. The recombinant adenovirus AdhTIMP-1 containing full-length cDNA of hTIMP-1 was generated by homologous recombination in E. coli. AdhTIMP-1 was then packaged and am- plified in adenoviral packaging cells , or human embryonic kidney 293 cells. The viral titer was checked by green fluo- rescent protein (GFP), and the expression of hTIMP-1 in vitro was detected by the techniques of Western blot and RT-PCR. RESULTS: The recombinant adenovir us vector carrying hTIMP-1 was constructed and confirmed by restriction en- donuclease analysis and DNA sequence analysis. The tran- scription of TIMP-1 mRNA in 293 cells was checked by RT-PCR and TIMP-1 protein could be detected in the cul - ture by Western blot analysis. CONCLUSION: The successful construction of recombi- nant adenoviral vector carrying human TIMP-1 and the ef-fective expression in vitro has laid a foundation for further study of its antitumor function and may pave the way for future application in liver gene therapy.