The large circular (LC)-antisense library to the 221 unigene clone was constructed and utilized in the identification of genes functionally involved in the growth of hepatocellular carcinoma cells. We identified that 37 out of the 221 members of the antisense library exerted a marked inhibitory effect on the growth of Huh-7. The putative functional categorization of each gene was then conducted on the basis of the sequence information. The relative expression levels of target genes were measured and treated with two LC-antisense molecules by real-time PCR. LC-antisense to EIF3EIP and AFP abolished the expression of EIF3EIP and AFP to the level of ≈7 and 39% compared to the control treatment in Huh-7 cells, respectively. LC-antisense molecules to EIF3EIP and AFP were simultaneously treated with 5-FU to Huh-7 cells. Two LC-antisense molecules showed additive effects with 5-FU compared with 5-FU alone, respectively. The combination of LC-antisense molecules and 5-FU showed a dramatic increase of sub-G1 apoptotic cell death fraction in cell cycle analysis, respectively. Therefore, these candidates may be used as target genes for drug development or adjuvant of conventional chemotherapeutic drugs.

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