RNAi-mediated knockdown of AURKB and EGFR shows enhanced therapeutic efficacy in prostate tumor regression

Abstract

Aurora B kinase (AURKB) and epidermal growth factor receptor 1 (EGFR) belong to serine/threonine and tyrosine kinase family of proteins. Both these kinases are found to overexpress in a large number of epithelial cancers, including hormonal refractory prostate cancer. In this communication, we present evidence for down-regulated expression of AURKB and EGFR, either alone or in combination, in prostate cancer cells. The results show that AURKB and EGFR were efficiently down-regulated in a dose-dependent manner. AURKB and EGFR siRNA in combination have shown enhanced therapeutic effect by inhibiting PC3 cell proliferation and inducing apoptosis in vitro, whereas androgen-dependent cancer cells LNCaP remain unaffected correlating the endogenous expression levels. Knockdown of AURKB and EGFR individually resulted in inhibition of prostate tumor growth in athymic nude mice and their combined knockdown resulted in tumor regression in which 40% of the treated mice were found to be tumor free, implicating the potential therapeutic benefits of AURKB-EGFR combination therapy.

VEGF-C differentially regulates VEGF-A expression in ocular and cancer cells; promotes angiogenesis via RhoA mediated pathway

“…Interferon response was detected by gene expression analysis using Interferon response detection kit (SI300A-1, SBI system Biosciences, Mountain view, CA, USA) following manufacturer's instructions. All the experiments were performed in triplicates, from independent transfections, and their average values (n = 3) were presented as detailed earlier [32].…”

siRNAs encapsulated in recombinant capsid protein derived from Dengue serotype 2 virus inhibits the four serotypes of the virus and proliferation of cancer cells

“…siRNA targeting an unrelated virus (Encephalomyocarditis virus, EMCV) was used as a negative control in all anti DEN viral assays. siRNAs targeting Rho A and AURK B were validated earlier for specific gene knockdown via the RNAi pathway and not by interferon stimulation (Addepalli et al, 2010;Kumar et al, 2011). In addition, a 27mer siRNA targeting EMCV was used as a negative control in all anti cancer assays.…”

“…DENV targeted siRNA cocktails contained 200 nM of each siRNA and equimolar concentration of the control siRNA was used in all antiviral experiments. 27mer siRNAs targeting AuRK B and Rho A, and the control siRNA targeting EMCV, were used at a pre-optimized concentration of 10 nM each as reported earlier (Addepalli et al, 2010;Kumar et al, 2011).…”

“…Evaluation of siRNA mediated knockdown of Aurorakinase B and RhoA was carried out by qRT-PCR, as described earler (Addepalli et al, 2010;Kumar et al, 2011), based on the qRTPCR relative quantitation method reported earlier (Kenneth and Thomas, 2001).…”

“…The DENV specific siRNAs that we have used in the study are targeted against conserved sites in the DENV genome. The anti cancer siRNAs that we have used have been reported earlier from our laboratory (Kumar et al, 2011;Addepalli et al, 2010;Chile et al, 2014). Furthermore, the reported yields of E. coli expressed flaviviral capsid proteins are extremely low (Jones et al, 2003).…”