Inhibitory Effects of HPV18-E6 Gene SiRNA on Growth of Hep-2 Cells

Objective: To investigate the interference efficiency of HPV18-E6 small interfering RNA (SiRNAs) in human laryn-geal carcinoma cancer cells line Hep-2. Methods: The HPV18-E6 gene SiRNA eukaryotic expression vector was constructed by inserting SiRNA cDNA into SiRNA insertion position of pSilencer 4. 1-svv vector. The HPV18-E6 SiRNA vector was transfected into HEP-2 cells line by lipofectamine 2000. The expression level of HPV18E6 mRNA was detected by using real-time quantitative reverse transcription polymerase chain reaction (real-time RT-PCR). The effect of HPV18-E6 SiRNA on cell cycle and apoptosis was determined by flow cy-tometry . The expression of HPV18-E6 SiRNA on Hep-2 cells was detected by Western blot. Results: We successfully designed expres-sion vector of SiRNA specifically targeting to HPV18-E6 mRNA. And the growth of Hep-2 cells was significantly suppressed by HPV18-E6 SiRNA. Before transfection, the expression of HPV18-E6 mRNA level was 1.04±0.28. However, the expression of HPV18-E6 mRNA level was 0.15±0.03 36 hours after transfection. The FCM shows the number of cells in G1 phase was increased after HPV18-E6 SiRNA transfection, which indicates the cell division was blocked in G1 pre-DNA-synthetic gap. Conclusion: The HPV18-E6 gene expression may play a important role in the growth of laryngeal carcinoma cells,and it might be a new target of the treatment for la-ryngeal carcinoma .