The combinational effect of E6/E7 siRNA and anti-miR-182 on apoptosis induction in HPV16-positive cervical cells

Abstract

In the present research, we assumed that reducing the amounts of E6 and E7 oncoproteins by a specific siRNA sequence and recovering p53 and RB proteins, along with the recovery of the FOXO1 protein by applying anti-miR-182, would increase apoptosis and reduce proliferation rate in cancer cells. The HPV16-positive CaSki cervical cancer cell line was used. 48 hours after transfection of siRNA for targeting E6 and E7 oncoproteins and anti-miR-182, expression of its cellular targets p53, p21 and FOXO1 was assessed by real-time PCR, western blot analysis and immunocytofluorescence staining. In all treatments, apoptosis rate and viability were evaluated using Annexin-V-FITC apoptosis detection kits and MTT assays, respectively. Among the designed siRNAs, E6-1 and E7-2 proved the most effective in reducing E6 and E7 expressions by increasing the apoptotic rates to 12.4% and 16%, respectively, after 48 hours. Also, using anti-miR-182 increased apoptotic rate to 12.7% 48 hours after transfection of cervical cancer cells. The combinational use of either E6-1 or E7-2 siRNAs with anti-miR-182 resulted in a rise in apoptosis to 19.3% and 26%, respectively, higher than those obtained from the individual application of either without anti-miR-182. The simultaneous use of siRNA E6-1 and siRNA E7-2 with cisplatin increased sensitivity to cisplatin and reduced the viability of the cancer cells as compared to the use of cisplatin alone. The simultaneous use of cisplatin and anti-miR-182 had no considerable effect on viability or apoptosis rate compared to cisplatin alone.

Inhibition of SF3b1 by pladienolide B evokes cycle arrest, apoptosis induction and p73 splicing in human cervical carcinoma cells

“…Cervical cancer is one of the most common cancers and the second leading cause of cancer-related death in women worldwide [1,2], with 274,000 deaths annually [3], which is also a complex disease involving the abnormal expression of many oncogenes and tumour suppressor genes [4]. The discovery of novel therapeutics requires a comprehensive understanding of the disease biology.…”

siRNA-E6 sensitizes HPV-16-related cervical cancer through Oxaliplatin: an in vitro study on anti-cancer combination therapy

“…In 2018, H. Javadi et al during a study on CaSki cancer cells showed that following combination therapy with E6-siRNA and cisplatin, the cells’ sensitivity to cisplatin and subsequent induction of apoptosis increases [ 19 ]. In the current study, the results of the MTT assay were also confirmed by flow cytometry assay with Annexin V/PI staining.…”

RIPOR2 Expression Decreased by HPV-16 E6 and E7 Oncoproteins: An Opportunity in the Search for Prognostic Biomarkers in Cervical Cancer

“…However, other studies evidence a controversy on the possible role of FOXO1 in cervical cancer, since its overexpression has been associated with a poor prognosis [ 57 ]. Moreover, it has been shown that the inhibition of the expression of E6 and E7 in Ca Ski cells recovers the expression of FOXO1, leading to apoptosis and to a reduction in the proliferation of cancer cells [ 58 ]. Interestingly, our RNAseq data showed a decrease in FOXO1 expression of −0.49 and −0.41 log2FC in cells with E6 and E7, respectively ( Tables S2 and S3 ).…”