Epoxyazadiradione, a neem-derived limonoid exhibits activities against pancreatic cancer through modulation of inflammatory molecules, lncRNAs, ROS, and EMT.

Pancreatic cancer is the seventh most prevalent cancer worldwide, with persistently low life expectancy that hasn't changed for many decades. Agents derived from Mother Nature has been emphasised for drug development due to their safety, affordability, and ability to modulate multiple cell signalling pathways. Among such agents, azadiradione (AZA) and epoxyazadiradione (EPA) are two limonoids derived from the neem plant (Azadirachta indica). Both of these limonoids are reported to exhibit cytotoxic activities against some cancer types. However, the potential of EPA and AZA against pancreatic cancer and the underlying mechanism has not been reported earlier. In this study, we investigated the potential of EPA and AZA against pancreatic cancer cells. Our results demonstrated that EPA possess greater cytotoxic potency against pancreatic cancer compared to AZA while non-malignant epithelial cells were only minimally affected by the limonoid. Further, EPA triggered apoptosis in pancreatic cancer cells as revealed by morphological changes, DNA fragmentation, cell cycle arrest, and annexin-V staining. EPA also induced ROS generation and altered the mitochondrial membrane potential. EPA inhibited the translocation of NF-κB-p65 and downregulated the expression of proteins related to cell survival (Bcl-xL, Bcl-2, survivin) and invasion (MMP-9), while inducing the proapoptotic protein (Bax). The expression of lncRNAs was modulated, and the gene silencing of GAS5 reversed the effects of EPA in pancreatic cancer lines. It also enhanced the sensitivity of cell lines to paclitaxel, decreased the spheroid formation and modulated the expressions of EMT markers. These cumulative findings revealed that EPA exhibits anti-cancer activity against pancreatic cancer.