Effect of antisense mRNA of human cervical cancer oncogene on the inhibition of growth in HepG2 Cells

Objective To investigate the effect of antisense nucleotide of human cervical cancer oncogene (HCCR) on the cell proliferation and apoptosis of HepG2 cells. Methods Antisense expression vector of pCDNA3.1-HCCR was constructed and transfected into HepG2 cells. The expression level of HCCR mRNA and protein were detected by real time PCR and Western blot. The effect of the pCDNA3.1-HCCR on cell proliferation was assayed by MTT method and the cell apoptosis was observed by flow cytometry. Results Real time PCR and Western blot analysis showed that antisense nucleotide of HCCR could effectively suppress HCCR expression level compared with those of non-transfected and the HCCR mRNA level at 24h after transfection was reduced to 16%, and the level of HCCR protein was also decreased .MTT method confirmed the cell proliferation of HepG2 cells were inhibited(P0.05), with the inhibition rate at 24h after transfection of 47.6%, while the apoptosis by flow cytometry was 1.74%±0.44%, 2.10%±0.30%, 14.34%±0.91% in HepG2 cells group, HepG2/pCDNA3.1(+) group and pCDNA3.1(+)-HCCR group, respectively , and the apoptosis of HepG2 cells transfected with pCDNA3.1(+)-HCCR increased much more than control groups(t=21.799,P0.05). Cell-cycle analysis by flow cytometry showed that transfected HepG2 cells were arrested at the G1 phase of the cell cycle. Conclusion The antisense nucleotide of HCCR can effectively inhibit the expression level of HCCR mRNA and protein , suppress cell proliferation and induce cell apoptosis and antisense nucleotide of HCCR may serve as a potential antitumor strategy in HCCs.