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Web End = Apoptosis (2015) 20:13881409 DOI 10.1007/s10495-015-1159-4
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Web End = In situ allicin generation using targeted alliinase deliveryfor inhibition of MIA PaCa-2 cells via epigenetic changes, oxidative stress and cyclin-dependent kinase inhibitor (CDKI) expression
Sagar V. Chhabria1 Mohammad A. Akbarsha2,3 Albert P. Li4 Prashant S. Kharkar5
Krutika B. Desai6
Published online: 19 August 2015 Springer Science+Business Media New York 2015
Abstract Allicin, an extremely active constituent of freshly crushed garlic, is produced upon reaction of substrate alliin with the enzyme alliinase (EC 4.4.1.4). Allicin has been shown to be toxic to several mammalian cells in vitro in a dose-dependent manner. In the present study this cytotoxicity was taken to advantage to develop a novel approach to cancer treatment, based on site directed generation of allicin. Alliinase was chemically conjugated to a monoclonal antibody (mAb) which was directed against a specic pancreatic cancer marker, CA19-9. After the CA19-9 mAb-alliinase conjugate was bound to targeted pancreatic cancer cells (MIA PaCa-2 cells), on addition of alliin, the cancer cell-localized alliinase produced allicin,
which effectively induced apoptosis in MIA PaCa-2 cells. Specicity of anticancer activity of in situ generated allicin was demonstrated using a novel in vitro systemintegrated discrete multiple organ co-culture technique. Further, allicin-induced caspase-3 expression, DNA fragmentation, cell cycle arrest, p21Waf1/Cip1 cyclin-dependent kinase inhibitor expression, ROS generation, GSH depletion, and led to various epigenetic modications which resulted in stimulation of apoptosis. This approach offers a new therapeutic strategy, wherein alliin and alliinase-bound antibody work together to produce allicin at targeted locations which would reverse gene silencing and suppress cancer cell growth, suggesting that combination of these targeted agents may improve pancreatic cancer therapy.
Keywords Allicin Apoptosis CA19-9 Cyclin-
dependent kinase inhibitor (CDKI) Integrated discrete
multiple organ co-culture (IdMOC) technique MIA PaCa-
2 cells
Introduction
Pancreatic adenocarcinoma is one of the most aggressive malignancies and has dismal prognosis. The overall 5-year survival rate of only 14 % is due to failure to detect this disease at an early stage, its aggressiveness, and the lack of effective conventional therapies [13]. The etiology...