Epoxyazadiradione Purified from the Azadirachta indica Seed Induced Mitochondrial Apoptosis and Inhibition of NFκB Nuclear Translocation in Human Cervical Cancer Cells

Abstract

Epoxyazadiradione (EAD) is an important limonoid present in Neem (Azadirachta indica) plant. In the present study, we have purified EAD from Neem seed and studied its anticancer potential in human cervical cancer (HeLa) cells. Cell proliferation inhibition studies indicated that the GI value of EAD is 7.5 ± 0.0092 μM in HeLa cells, whereas up to 50 μM concentrations EAD did not affect the growth of normal H9C2 cells. The control drug cisplatin inhibited the growth of both HeLa and H9C2 cells with a GI value of 2.92 ± 1.192 and 4.22 ± 1.568 μM, respectively. Nuclear DNA fragmentation, cell membrane blebbing, phosphatidylserine translocation, upregulation of Bax, caspase 3 activity and poly (ADP ribose) polymerase cleavage and downregulation of BCl2 in HeLa cells on treatment with EAD indicated the apoptotic cell death. Increase in caspase 9 activity and release of active cytochrome c to the cytoplasm on treatment with EAD confirmed that the apoptosis was mediated through the mitochondrial pathway. Epoxyazadiradione also inhibited the nuclear translocation of nuclear factor κB in HeLa cells. Thus, our studies demonstrated EAD as a potent and safe chemotherapeutic agent when compared with the standard drug cisplatin that is toxic to both cancer and normal cells equally. Copyright © 2017 John Wiley & Sons, Ltd.

A comprehensive review of phytochemical profile, bioactives for pharmaceuticals, and pharmacological attributes of Azadirachta indica

“…Increase in activity of caspase IX and release of active cytochrome-c with inhibition of nuclear translocation factor in HeLa cells was noted. The results were comparable to standard drug, cis-platin which suggested epoxyazadiradione as an alternative chemotherapeutic agent (Shilpa et al, 2017). Seed extract showed significant activity against chronic and acute inflammatory diseases.…”

Nimbolide induces cell death in T lymphoma cells: Implication of altered apoptosis and glucose metabolism

“…It is isolated from the leaves and flowers of a traditional medicinal plant, Azadirachta indica (Neem). Besides nimbolide, other limonoids such as azadirachtin, nimbidin, and epoxyazadiradione have also shown various beneficial therapeutic activities, including anti‐inflammatory, and anticancer 3‐11 . Notably, available reports suggest that among all the limonoids, nimbolide is the most potent anticancer agent 12 …”

“…Besides nimbolide, other limonoids such as azadirachtin, nimbidin, and epoxyazadiradione have also shown various beneficial therapeutic activities, including anti-inflammatory, and anticancer. [3][4][5][6][7][8][9][10][11] Notably, available reports suggest that among all the limonoids, nimbolide is the most potent anticancer agent. 12 Numerous in vitro and in vivo studies have demonstrated the promising therapeutic efficacy of nimbolide against breast, colon, prostate, pancreatic, cervical, renal, and oral cancers.…”

Epoxyazadiradione induced apoptosis/anoikis in triple‐negative breast cancer cells, MDA‐MB‐231, by modulating diverse cellular effects

“…Isolation and characterization of EAD were done as described in our previous report. [15] Briefly, EAD was isolated from the neem seed cake using repeated petroleum ether cum acetone extraction, chromatography, and crystallization process to yield 1.895 g/kg of cake. The compound was later characterized by comparing its spectroscopic data (Fourier-transform infrared spectroscopy, 1 H nuclear magnetic resonance [ 1 HNMR], 13 C NMR) and high-performance liquid chromatography profile with the existing literature.…”

“…Previously, we have reported the purification of EAD from the petroleum ether extract of neem seeds and their anticancer effects/ mechanism in human cervical cancer cells. [15] In the present paper, Fluorometric Assay Kit (BioVision).…”