, Volume 25, Issue 1, pp 88-92
Date: 09 Aug 2007

Inhibitory effect of Hypoxia inducible factor-1 antisense oligonucleotide on growth of human hepatocellular carcinoma cells

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Abstract

Object

To observe the inhibitory effect of Hypoxia inducible factor-1 antisense oligonuclecotide on growth of human hepatocellular carcinoma cells and gene expression of HIF-1, in order to seek a new gene therapy for hepatocellular carcinoma.

Methods

Six Hypoxia inducible factor-1 antisense oligonuclecotides with various concentrations (0.2 μmol/l, 0.4 μmol/l, and 0.8 μmol/l) were transformed into HepG2 cells by lipofectamine reagent. 72 h after transfection, MTS assay was used to detect cellular proliferation. In addition, Hypoxia inducible factor-1 antisense oligonuclecotide2 with various concentrations (0.2, 0.4, 0.8, and 1.0 μmol/l) were transformed into HepG2 cells. About 48 h after transfection, reverse transcription-polymerase chain reaction assay and Western Blot assay were employed to detect the expression of Hypoxia inducible factor-1 gene and the synthesis of Hypoxia inducible factor-1 protein respectively.

Results

HepG2 cell growth was inhibited by 6 Hypoxia inducible factor-1 antisense oligonuclecotides at various concentrations. Among them, Hypoxia inducible factor-1 antisense oligonuclecotide2 showed the most effective inhibition ability (P < 0.01), the inhibitory rate was 89.66% at the concentration of 1.0 μmol/l. About 48 h after transfection, Hypoxia inducible factor-1 mRNA expression was downregulated and Hypoxia inducible factor-1 protein synthesis was decreased by antisense oligonuclecotide2.

Conclusions

The hepatocellular carcinoma cell proliferation was inhibited by Hypoxia inducible factor-1 antisense oligonuclecotide. Moreover, the gene expression and protein synthesis of Hypoxia inducible factor-1 were reduced by Hypoxia inducible factor-1 antisense oligonuclecotide. The findings suggested that antisense technique targeting Hypoxia inducible factor-1 might be an effective gene therapy of human hepatocellular carcinoma.