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AIM: To investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis in human hepatoceliular carcinoma (HCC) cell line HepG2 and normal liver cell line L02. METHODS: We constructed the recombinant replication-incompetent Ad.mda-7 virus vector and infected it into the human HCC cell line HepG2 and normal liver cell line L02. RT-PCR was performed to detect the mRNA expressing in cells. by ELISA was used to detect MDA-7/ IL-24 protein expression in the culture supernatant. The effect of apoptosis induced by Ad.mda-7 was confirmed by Hoechst staining and flow cytometry assay with An-nexin-V and PI staining. MTT assay was used to determine growth inhibition of HepG2 cells, and cell-cycle and hypodiploidy analyses were performed by flow cytometry. RESULTS: Recombinant replication-defective virus expressing MDA-7/IL-24 was constructed successfully. RT-PCR showed that the Ad.mda-7 could mediate the expression of the exogenous gene MDA-7/IL-24 into HepG2 and L02. The concentration of MDA-7/IL-24 protein in supernatant was 130 pg/mL and 110 pg/mL in Ad.mda-7-infected L02 and HepG2 cells, respectively. Ad.mda-7 infection obviously induced apoptosis (from 2.60±0.72% to 33.6±13.2%, P= 0.00012) and growth suppression in HepG2 (inhibition ratio IR = 68%) and an increase in the percentage of specific cancer cell types at the G2/M phase of the cell cycle (from 6.44% to 32.29%, P<0.01), but not in L02 cells. CONCLUSION: These results confirm selectively induction of apoptosis and growth suppression by the mda-7/ IL-24 gene with replication-incompetent adenovirus vector in human hepatocellular carcinoma cell line HepG2.
AIM: To investigate the effect of replication-incompetent adenovirus vector expressing MDA-7 / IL-24 on tumor growth and apoptosis in human hepatoceliular carcinoma (HCC) cell line HepG2 and normal liver cell line L02. METHODS: We constructed the recombinant replication- incompetent Ad.mda-7 virus vector and infected it into the human HCC cell line HepG2 and normal liver cell line L02. RT-PCR was performed to detect the mRNA expressing in cells. by ELISA was used to detect MDA-7 / IL- 24 protein expression in the culture supernatant. The effect of apoptosis induced by Ad.mda-7 was confirmed by Hoechst staining and flow cytometry assay with An-nexin-V and PI staining. MTT assay was used to determine growth inhibition of HepG2 cells, RESULTS: Recombinant replication-defective virus expressing MDA-7 / IL-24 was constructed successfully. RT-PCR showed that the Ad.mda-7 could mediate the expression of the exogenous gene MDA -7 / IL-24 into HepG2 and L02. The concentration of MDA-7 / IL-24 protein in supernatant was 130 pg / mL and 110 pg / mL in Ad.mda-7-infected L02 and HepG2 cells, respectively. Ad inhibition of induced apoptosis (from 2.60 ± 0.72% to 33.6 ± 13.2%, P = 0.00012) and growth inhibition in HepG2 (inhibition ratio IR = 68%) and an increase in the percentage of specific cancer cell types at the CONCLUSION: These results confirm selectively induction of apoptosis and growth suppression by the mda-7 / IL-24 gene with G2 / M phase of the cell cycle (from 6.44% to 32.29%, P <0.01) replication-incompetent adenovirus vector in human hepatocellular carcinoma cell line HepG2.