Expression of a dominant negative inhibitor of NF-kappaB protects MIN6 beta-cells from cytokine-induced apoptosis

The Journal of Surgical Research
M S BakerDixon B Kaufman

Abstract

Islet graft injury by cytokines released from inflammatory cells (macrophages) that infiltrate the transplant site is an important mechanism of early islet transplant dysfunction. This detrimental "cytokine effect" is thought to be mediated by NF-kappaB-dependent up-regulation of iNOS gene expression and increased nitric oxide (NO) production by the islet. We attempted to make a beta-cell resistant to cytokine-induced apoptosis by transfecting the parent line with a dominant negative inhibitor of NF-kappaB. A flag-tagged IkappaBalphaM cDNA subcloned into an SFFV-neo vector was used to transfect parent beta-Cell line MIN6. MIN6 and the resultant mutant (2Bm) were cultured for 24 h in a cytokine mixture including IL-1beta (50 units/mL), TNF-alpha (1000 units/mL), and IFN-gamma (750 units/mL) and cotreated with either the iNOS inhibitor L-NMMA (1 mM) or the caspase inhibitor Z-VAD (0.1 mM). NF-kappaB translocation was determined by gel shift. Nitrite production was determined by the Griess reaction. Apoptosis was determined by flow cytometry. When treated with cytokine 2Bm demonstrated significantly less NF-kappaB nuclear translocation, nitrite production, and apoptosis than parent MIN6. The rate of apoptosis in cytokine-treated 2...Continue Reading

References

Jun 1, 1984·Transplantation·R W SchwizerR Evans
Jan 1, 1994·Annual Review of Cell Biology·U SiebenlistK Brown
Sep 30, 1993·The New England Journal of Medicine·UNKNOWN Diabetes Control and Complications Trial Research GroupC Siebert
Mar 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·J A CorbettM L McDaniel
Apr 1, 1997·Seminars in Cancer Biology·G E Sonenshein
May 23, 1998·Annual Review of Immunology·S GhoshE B Kopp
Jun 8, 1999·Annual Review of Immunology·J C Rathmell, C B Thompson
Oct 19, 1999·The Journal of Experimental Medicine·S T GreyC Ferran
Feb 15, 2000·Annals of the New York Academy of Sciences·A Bal-PriceG C Brown
Feb 28, 2002·Science's STKE : Signal Transduction Knowledge Environment·D M Rothwarf, M Karin

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Citations

Mar 27, 2009·Apoptosis : an International Journal on Programmed Cell Death·Helen E ThomasThomas W Kay
Nov 16, 2004·Molecular and Cellular Biology·Jiang-Yan YangChristian Widmann
Mar 23, 2006·Proceedings of the National Academy of Sciences of the United States of America·R EldorD Melloul
Feb 3, 2007·Proceedings of the National Academy of Sciences of the United States of America·Sunshin KimMyung-Shik Lee
Oct 10, 2009·Pharmacological Reports : PR·Sandip Patel, Dev Santani
Mar 5, 2013·Biochemical and Biophysical Research Communications·Akira OgasawaraKazuo Umezawa
Jul 20, 2010·The Journal of Surgical Research·Tohru TakahashiSatoru Todo
Dec 18, 2003·Annals of the New York Academy of Sciences·Decio L EizirikAlessandra K Cardozo
Jan 26, 2006·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·D YinA S Chong
Oct 25, 2011·Transplant International : Official Journal of the European Society for Organ Transplantation·Xiao-Hong WangJin Zheng
Feb 20, 2007·Pharmacology & Therapeutics·Máire E Doyle, Josephine M Egan
Feb 27, 2017·Trends in Cell Biology·Leen Catrysse, Geert van Loo
Jun 3, 2004·Journal of Cellular Physiology·Hongxiang HuiRiccardo Perfetti
Apr 11, 2018·Journal of Molecular Endocrinology·Tarlliza R NardelliFernanda Ortis
Aug 16, 2013·The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology·Cha Kyung YounYoung Jin Jeon
Jul 25, 2007·Diabetes, Obesity & Metabolism·Michael DeckerAlan N Elias
Dec 7, 2007·Journal of Cellular and Molecular Medicine·L SusickA Kowluru
Aug 8, 2009·Journal of Huazhong University of Science and Technology. Medical Sciences = Hua Zhong Ke Ji Da Xue Xue Bao. Yi Xue Ying De Wen Ban = Huazhong Keji Daxue Xuebao. Yixue Yingdewen Ban·Yuzhen LiangLeping Feng
Jan 7, 2006·Diabetes/metabolism Research and Reviews·Cillian McCabeTimothy O'Brien
Dec 6, 2002·Drugs·Nikolai PetrovskyDesmond A Schatz
Mar 22, 2006·Pediatric Research·Juris J MeierPeter C Butler
Feb 24, 2010·Der Orthopäde·K-D ThomannM Rauschmann
Jul 18, 2014·Endocrine Journal·Atsushi YoshikawaIichiro Shimomura
Aug 22, 2014·Biomolecules & Therapeutics·Eun Jeong KimYoung Jin Jeon
Oct 28, 2019·Journal of Clinical Medicine·Hirofumi Noguchi
May 17, 2008·Biochemical Society Transactions·Danielle Melloul
Jan 22, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Yangming XiaoGuangming Zhong
Apr 17, 2003·The Journal of Surgical Research·Marshall S BakerDixon B Kaufman

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