Sulforaphane-induced apoptosis involves the type 1 IP3 receptor

In this study we show that anti-tumor effect of sulforaphane (SFN) is partially realized through the type 1 inositol 1,4,5-trisphosphate receptor (IP3R1). This effect was verified in vitro on three different stable cell lines and also in vivo on the model of nude mice with developed tumors. Early response (6 hours) of A2780 ovarian carcinoma cells to SFN treatment involves generation of mitochondrial ROS and increased transcription of NRF2 and its downstream regulated genes including heme oxygenase 1, NAD(P)H:quinine oxidoreductase 1, and KLF9. Prolonged SFN treatment (24 hours) upregulated expression of NRF2 and IP3R1. SFN induces a time-dependent phosphorylation wave of HSP27. Use of IP3R inhibitor Xestospongin C (Xest) attenuates both SFN-induced apoptosis and the level of NRF2 protein expression. In addition, Xest partially attenuates anti-tumor effect of SFN in vivo. SFN-induced apoptosis is completely inhibited by silencing of IP3R1 gene but only partially blocked by silencing of NRF2; silencing of IP3R2 and IP3R3 had no effect on these cells. Xest inhibitor does not significantly modify SFN-induced increase in the rapid activity of ARE and AP1 responsive elements. We found that Xest effectively reverses the SFN-dependent increase of nuclear content and decrease of reticular calcium content. In addition, immunofluorescent staining with IP3R1 antibody revealed that SFN treatment induces translocation of IP3R1 to the nucleus. Our results clearly show that IP3R1 is involved in SFN-induced apoptosis through the depletion of reticular calcium and modulation of transcription factors through nuclear calcium up-regulation.

[1]  M. Ananthanarayanan,et al.  Nuclear Factor, Erythroid 2-Like 2 Regulates Expression of Type 3 Inositol 1,4,5-Trisphosphate Receptor and Calcium Signaling in Cholangiocytes. , 2015, Gastroenterology.

[2]  Kisha Mitchell,et al.  Nuclear Factor Erythroid 2-like 2 Regulates Expression of Inositol 1,4,5-trisphosphate Receptor, Type 3 and Calcium Signaling in Cholangiocytes , 2015 .

[3]  D. Cholujová,et al.  Modulation of cisplatin sensitivity in human ovarian carcinoma A2780 and SKOV3 cell lines by sulforaphane. , 2014, Toxicology letters.

[4]  D. Llères,et al.  Monitoring Keap1–Nrf2 interactions in single live cells , 2014, Biotechnology advances.

[5]  Yung-Hyun Choi,et al.  Sulforaphane induces apoptosis in T24 human urinary bladder cancer cells through a reactive oxygen species-mediated mitochondrial pathway: the involvement of endoplasmic reticulum stress and the Nrf2 signaling pathway. , 2014, International journal of oncology.

[6]  L. Missiaen,et al.  Inositol 1,4,5-trisphosphate receptor-isoform diversity in cell death and survival. , 2014, Biochimica et biophysica acta.

[7]  D. Traver,et al.  Loss of IP3R-dependent Ca2+ signalling in thymocytes leads to aberrant development and acute lymphoblastic leukemia , 2014, Nature Communications.

[8]  S. Srivastava,et al.  Molecular targets of isothiocyanates in cancer: recent advances. , 2014, Molecular nutrition & food research.

[9]  Rivka L. Shoulson,et al.  Resveratrol inhibits estrogen-induced breast carcinogenesis through induction of NRF2-mediated protective pathways. , 2014, Carcinogenesis.

[10]  A. Kong,et al.  Apigenin Reactivates Nrf2 Anti-oxidative Stress Signaling in Mouse Skin Epidermal JB6 P + Cells Through Epigenetics Modifications , 2014, The AAPS Journal.

[11]  Conceição Santos,et al.  Sulforaphane Induces Oxidative Stress and Death by p53-Independent Mechanism: Implication of Impaired Glutathione Recycling , 2014, PloS one.

[12]  Jianmin Wang,et al.  Nrf2 amplifies oxidative stress via induction of Klf9. , 2014, Molecular cell.

[13]  S. Pervaiz,et al.  Small molecule sensitization to TRAIL is mediated via nuclear localization, phosphorylation and inhibition of chaperone activity of Hsp27 , 2013, Cell Death and Disease.

[14]  J. Sedlák,et al.  Sulphide signalling potentiates apoptosis through the up‐regulation of IP3 receptor types 1 and 2 , 2013, Acta physiologica.

[15]  K. Mikoshiba,et al.  IP3R2 levels dictate the apoptotic sensitivity of diffuse large B-cell lymphoma cells to an IP3R-derived peptide targeting the BH4 domain of Bcl-2 , 2013, Cell Death and Disease.

[16]  D. Dobrota,et al.  Effect of aging on formation of reactive oxygen species by mitochondria of rat heart. , 2013, General physiology and biophysics.

[17]  Kyong-Tai Kim,et al.  Apoptotic Phosphorylation of Histone H3 on Ser-10 by Protein Kinase Cδ , 2012, PloS one.

[18]  L. Ignarro,et al.  Sensing endoplasmic reticulum stress by protein kinase RNA‐like endoplasmic reticulum kinase promotes adaptive mitochondrial DNA biogenesis and cell survival via heme oxygenase‐1/carbon monoxide activity , 2012, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[19]  O. Krizanova,et al.  IP(3) receptors, stress and apoptosis. , 2012, General physiology and biophysics.

[20]  Yi-Chiang Hsu,et al.  Effect of Sulforaphane on Growth Inhibition in Human Brain Malignant Glioma GBM 8401 Cells by Means of Mitochondrial- and MEK/ERK-Mediated Apoptosis Pathway , 2012, Cell Biochemistry and Biophysics.

[21]  J. McCarron,et al.  Mitochondrial regulation of cytosolic Ca2+ signals in smooth muscle , 2012, Pflügers Archiv - European Journal of Physiology.

[22]  S. Pastoreková,et al.  Apoptosis induced clustering of IP3R1 in nuclei of non‐differentiated PC12 cells , 2011, Journal of cellular physiology.

[23]  L. Missiaen,et al.  IP3 Receptors, Mitochondria, and Ca2+ Signaling: Implications for Aging , 2011, Journal of aging research.

[24]  Alexander D. MacKerell,et al.  Characterization of ERK Docking Domain Inhibitors that Induce Apoptosis by Targeting Rsk-1 and Caspase-9 , 2011, BMC Cancer.

[25]  S. Jang,et al.  Sulforaphane suppresses TARC/CCL17 and MDC/CCL22 expression through heme oxygenase-1 and NF-κB in human keratinocytes , 2010, Archives of pharmacal research.

[26]  C. Garrido,et al.  Sulforaphane Activates Heat Shock Response and Enhances Proteasome Activity through Up-regulation of Hsp27* , 2010, The Journal of Biological Chemistry.

[27]  G. Fiskum,et al.  Neuroprotection through stimulation of mitochondrial antioxidant protein expression. , 2010, Journal of Alzheimer's disease : JAD.

[28]  Jian Peng,et al.  Induction of Ca2+ signal mediated apoptosis and alteration of IP3R1 and SERCA1 expression levels by stress hormone in differentiating C2C12 myoblasts. , 2010, General and comparative endocrinology.

[29]  A. Zahradníková,et al.  Type 2 IP(3) receptors are involved in uranyl acetate induced apoptosis in HEK 293 cells. , 2009, Toxicology.

[30]  Paul Talalay,et al.  Direct and indirect antioxidant properties of inducers of cytoprotective proteins. , 2008, Molecular nutrition & food research.

[31]  S. Yoo,et al.  Presence of secretogranin II and high-capacity, low-affinity Ca2+ storage role in nucleoplasmic Ca2+ store vesicles. , 2007, Biochemistry.

[32]  Jing Zhao,et al.  Enhancing Expression of Nrf2-Driven Genes Protects the Blood–Brain Barrier after Brain Injury , 2007, The Journal of Neuroscience.

[33]  G. Hajnóczky,et al.  IP3 receptors in cell survival and apoptosis: Ca2+ release and beyond , 2007, Apoptosis.

[34]  Shyam Biswal,et al.  Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway. , 2007, Annual review of pharmacology and toxicology.

[35]  A. Kong,et al.  Gene expression profiles induced by cancer chemopreventive isothiocyanate sulforaphane in the liver of C57BL/6J mice and C57BL/6J/Nrf2 (-/-) mice. , 2006, Cancer letters.

[36]  A. Kong,et al.  Cancer chemoprevention of intestinal polyposis in ApcMin/+ mice by sulforaphane, a natural product derived from cruciferous vegetable. , 2006, Carcinogenesis.

[37]  D. Mueller,et al.  p300/Cyclic AMP-Responsive Element Binding-Binding Protein Mediates Transcriptional Coactivation by the CD28 T Cell Costimulatory Receptor1 , 2006, The Journal of Immunology.

[38]  D. Sawyer,et al.  Heat shock proteins in cancer: chaperones of tumorigenesis. , 2006, Trends in biochemical sciences.

[39]  J. Jakubikova,et al.  Effect of isothiocyanates on nuclear accumulation of NF-kappaB, Nrf2, and thioredoxin in caco-2 cells. , 2006, Journal of agricultural and food chemistry.

[40]  Moira A. Taylor,et al.  Glutathione S-transferase M1 polymorphism and metabolism of sulforaphane from standard and high-glucosinolate broccoli. , 2005, The American journal of clinical nutrition.

[41]  M. Mattson,et al.  Suppression of Calcium Release from Inositol 1,4,5-Trisphosphate-sensitive Stores Mediates the Anti-apoptotic Function of Nuclear Factor-κB* , 2005, Journal of Biological Chemistry.

[42]  A. Kong,et al.  Suppression of NF-kappaB and NF-kappaB-regulated gene expression by sulforaphane and PEITC through IkappaBalpha, IKK pathway in human prostate cancer PC-3 cells. , 2005, Oncogene.

[43]  M. Bootman,et al.  Cell Signalling: IP3 Receptors Channel Calcium into Cell Death , 2004, Current Biology.

[44]  J. Goris,et al.  Caspase-3-induced Truncation of Type 1 Inositol Trisphosphate Receptor Accelerates Apoptotic Cell Death and Induces Inositol Trisphosphate-independent Calcium Release during Apoptosis* , 2004, Journal of Biological Chemistry.

[45]  P. Karplus,et al.  A Novel Mechanism of Chemoprotection by Sulforaphane , 2004, Cancer Research.

[46]  H. Beere.  `The stress of dying': the role of heat shock proteins in the regulation of apoptosis , 2004, Journal of Cell Science.

[47]  K. Singletary,et al.  Sulforaphane: a naturally occurring mammary carcinoma mitotic inhibitor, which disrupts tubulin polymerization. , 2003, Carcinogenesis.

[48]  R. Dhir,et al.  Sulforaphane induces caspase-mediated apoptosis in cultured PC-3 human prostate cancer cells and retards growth of PC-3 xenografts in vivo. , 2003, Carcinogenesis.

[49]  S. Biswal,et al.  Identification of Nrf2-regulated genes induced by the chemopreventive agent sulforaphane by oligonucleotide microarray. , 2002, Cancer research.

[50]  F. Chung,et al.  Isothiocyanates as cancer chemopreventive agents: their biological activities and metabolism in rodents and humans. , 2002, Current drug metabolism.

[51]  H. Bartsch,et al.  Nuclear Factor κB Is a Molecular Target for Sulforaphane-mediated Anti-inflammatory Mechanisms* , 2001, The Journal of Biological Chemistry.

[52]  R. Hipskind,et al.  Induction-independent Recruitment of CREB-binding Protein to the c-fos Serum Response Element through Interactions between the Bromodomain and Elk-1* , 2001, The Journal of Biological Chemistry.

[53]  H. Bartsch,et al.  Nuclear factor kappa B is a molecular target for sulforaphane-mediated anti-inflammatory mechanisms. , 2001, The Journal of biological chemistry.

[54]  Y. Zhang,et al.  Role of glutathione in the accumulation of anticarcinogenic isothiocyanates and their glutathione conjugates by murine hepatoma cells. , 2000, Carcinogenesis.

[55]  R. Kraft,et al.  MAPKAP Kinase 2 Phosphorylates Serum Response Factor in Vitro and in Vivo* , 1999, The Journal of Biological Chemistry.

[56]  J. Fahey,et al.  Broccoli sprouts: an exceptionally rich source of inducers of enzymes that protect against chemical carcinogens. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[57]  C. Cho,et al.  A major inducer of anticarcinogenic protective enzymes from broccoli: isolation and elucidation of structure. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[58]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.