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Hepatocellular carcinoma is a lethal malignancy with poor prognosis, partially due to tumor metastasis, recurrence and resistance to chemo- or radio-therapy. Cisplatin can inhibit cancer cell DNA replication, and is widely used in the clinical treatment of tumors. The present study aimed to generate eukaryotic expression vectors for Livin shRNA and to examine the effects of Livin shRNA on the chemosensitivity of HepG2 hepatocellular carcinoma cells. Eukaryotic expression vectors for Livin shRNA (pSD11-U6/Neo/GFP/Livin) were designed and constructed. The HepG2 hepatocellular carcinoma cell line was transfected with this vector using the liposome method. The expression levels of Livin mRNA and protein were measured by quantitative polymerase chain reaction and western blot analysis. The rate of cell growth inhibition was measured using MTT assay following treatment of the cells with cisplatin (2.0 mg/l). DNA sequencing confirmed that the construction of the eukaryotic expression vector for Livin shRNA had been successful. Transfection of these vectors into HepG2 cells led to a significant reduction in the expression levels of Livin mRNA and protein (P<0.05). Cisplatin treatment was associated with significantly higher rates of cell growth inhibition in HepG2 cells transfected with Livin shRNA compared with those that were not transfected (P<0.05). The vectors constructed in the present study produced effective inhibition of the Livin gene in HepG2 cells and increased the chemosensitivity of hepatocellular carcinoma cells. [ABSTRACT FROM AUTHOR]