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In situ allicin generation using targeted alliinase delivery for inhibition of MIA PaCa-2 cells via epigenetic changes, oxidative stress and cyclin-dependent kinase inhibitor (CDKI) expression

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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 specific 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. Specificity of anticancer activity of in situ generated allicin was demonstrated using a novel in vitro system—integrated 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 modifications 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.

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Acknowledgments

This work was supported in part by Mumbai University Grant (097). We appreciate Sophisticated Analytical Instrumentation Facility, Indian Institute of Technology, Bombay, for Flow cytometry analysis, BD Biosciences for providing Matrigel, and Dr. Vibha Verma for technical assistance.

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Authors and Affiliations

  1. Department of Biological Sciences, School of Science, SVKM’s NMIMS University, Vile Parle (W), Mumbai, 400056, India

    Sagar V. Chhabria

  2. Mahatma Gandhi-Doerenkamp Center (MGDC) for Alternatives, Bharathidasan University, Tiruchirappalli, Tamil Nadu, 620024, India

    Mohammad A. Akbarsha

  3. Department of Food Science and Nutrition, College of Food and Agriculture, King Saud University, Riyadh, 12372, Kingdom of Saudi Arabia

    Mohammad A. Akbarsha

  4. In Vitro ADMET Laboratories LLC, 9221 Rumsey Road, Suite 8, Columbia, MD, 21045, USA

    Albert P. Li

  5. Department of Pharmaceutical Chemistry, Shobhaben Pratapbhai Patel School of Pharmacy and Technology Management, SVKM’s NMIMS University, Vile Parle (W), Mumbai, 400056, India

    Prashant S. Kharkar

  6. Department of Microbiology, SVKM’s Mithibai College of Arts, Chauhan Institute of Science & Amrutben Jivanlal College of Commerce & Economics, Vile Parle (W), Mumbai, 400056, India

    Krutika B. Desai

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  1. Sagar V. Chhabria
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Correspondence to Krutika B. Desai.

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Dr. Albert P. Li is the inventor of IdMOC and CEO of AP Sciences Inc., Columbia, MD, USA which commercializes IdMOC culture plates.

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Chhabria, S.V., Akbarsha, M.A., Li, A.P. et al. In situ allicin generation using targeted alliinase delivery for inhibition of MIA PaCa-2 cells via epigenetic changes, oxidative stress and cyclin-dependent kinase inhibitor (CDKI) expression. Apoptosis 20, 1388–1409 (2015). https://doi.org/10.1007/s10495-015-1159-4

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