Abstract 2115: Effect of small interfering RNA targeting HPV E6/E7 gene on the regulation of TP53/Rb dynamic behaviour in cervical cancer cells

Human papillomavirus (HPV) E6 and E7 viral oncogenes are very well known to cause cervical cancer, because E6 degrades TP53 tumor suppressor protein, and E7 inactivates the tumor suppressor retinoblastoma (pRb) protein. Thus E6 and E7 oncogenes of HPV are supposed to be promising targets of gene therapy against HPV mediated cervical cancer. Here, we attempted to study the regulation of TP53/pRb proteins dynamic behaviour after HPV E6/E7 small interfering RNA (siRNA) transfection in cervical cancer cells. HPV positive (HeLa and Caski) cell lines were selected for these experiments. Herein, we also validated the dynamics of TP53 in response to antiviral siRNA targeting the E6 and E7 oncogenes. At first, we analyzed the effect of HPV E6/E7 siRNA on TP53/pRb reporter gene activity by using pathway profiling luciferase system. Our result clearly indicates that when compared with E6/E7 siRNA pool alone, siRNA pool in combination cisplatin dramatically increases the TP53/pRb and decreases E2F response elements regulating luciferase reporter gene activity. TP53 protein and its target gene expression were also confirmed by western blot and qRT-PCR analysis in a time dependent manner. In our TP53 reporter gene (pGreenFire1 GFP reporter vector with EF1-puro) assay by using live imaging analysis shows that combination of HPVE6/E7 siRNA pool with cisplatin, induces the TP53 expression within 10h and sustained that leads to cell death. Our imaging results also suggested that combining cisp...
Source: Cancer Research - Category: Cancer & Oncology Authors: Tags: Molecular and Cellular Biology Source Type: research