Abrogation of HPV-induced cancer phenotypes by shRNA against HPV E6, E7 and A3B.

<p>(a) A3B mRNA level in stable A3B, E6 or E7-knockdown MCF10A-HPV18 cells. A3B, E6 or E7 stably knocked down cells were established using shRNA retroviral vector. Total RNA was extracted from cells and then subjected to qRT-PCR. (b) γH2AX level in stable A3B-knockdown MCF10A-HPV18 cells. Cells were lysed after selection using puromycin and then subjected to western blot. The γH2AX and β-actin were detected using anti-γH2AX and β-actin Ab. The number at bottom of panel shows band intensity ratio after normalized by β-actin level. (c and d) HPV-18 E6 or E7 mRNA level in stable E6 or E7-knockdown MCF10A-HPV18 cells. Total RNA was extracted and then subjected to qRT-PCR. (e) γH2AX level in stable HPV18 E6 or E7-knockdown MCF10A- HPV18 cells. Cells were lysed after selection using puromycin, and then subjected to western blot. The γH2AX and β-actin were detected using anti-γH2AX and β-actin Ab. The number at bottom of panel shows band intensity ratio after normalization by β-actin level. Values represent the mean ± SD of at least three independent experiments. Student’s t-test was performed, with <i>p</i>-values indicated on the graph.