Cellular and Molecular Life Sciences

, Volume 76, Issue 3, pp 609–625 | Cite as

Endoplasmic reticulum-targeting doxorubicin: a new tool effective against doxorubicin-resistant osteosarcoma

  • Ilaria Buondonno
  • Elena Gazzano
  • Elisa Tavanti
  • Konstantin Chegaev
  • Joanna Kopecka
  • Marilù Fanelli
  • Barbara Rolando
  • Roberta Fruttero
  • Alberto Gasco
  • Claudia Hattinger
  • Massimo Serra
  • Chiara RigantiEmail author
Original Article


Doxorubicin is one of the most effective drugs for the first-line treatment of high-grade osteosarcoma. Several studies have demonstrated that the major cause for doxorubicin resistance in osteosarcoma is the increased expression of the drug efflux transporter ABCB1/P-glycoprotein (Pgp). We recently identified a library of H2S-releasing doxorubicins (Sdox) that were more effective than doxorubicin against resistant osteosarcoma cells. Here we investigated the molecular mechanisms of the higher efficacy of Sdox in human osteosarcoma cells with increasing resistance to doxorubicin. Differently from doxorubicin, Sdox preferentially accumulated within the endoplasmic reticulum (ER), and its accumulation was only modestly reduced in Pgp-expressing osteosarcoma cells. The increase in doxorubicin resistance was paralleled by the progressive down-regulation of genes of ER-associated protein degradation/ER-quality control (ERAD/ERQC), two processes that remove misfolded proteins and protect cell from ER stress-triggered apoptosis. Sdox, that sulfhydrated ER-associated proteins and promoted their subsequent ubiquitination, up-regulated ERAD/ERQC genes. This up-regulation, however, was insufficient to protect cells, since Sdox activated ER stress-dependent apoptotic pathways, e.g., the C/EBP-β LIP/CHOP/PUMA/caspases 12-7-3 axis. Sdox also promoted the sulfhydration of Pgp that was subsequently ubiquitinated: this process further enhanced Sdox retention and toxicity in resistant cells. Our work suggests that Sdox overcomes doxorubicin resistance in osteosarcoma cells by at least two mechanisms: it induces the degradation of Pgp following its sulfhydration and produces a huge misfolding of ER-associated proteins, triggering ER-dependent apoptosis. Sdox may represent the prototype of innovative anthracyclines, effective against doxorubicin-resistant/Pgp-expressing osteosarcoma cells by perturbing the ER functions.


Osteosarcoma P-glycoprotein H2S-releasing doxorubicin Endoplasmic reticulum-associated protein degradation Endoplasmic reticulum stress 





ATP binding cassette B1/P-glycoprotein


Hydrogen sulfide


Reactive oxygen species


H2S-releasing doxorubicin


Fetal bovine serum


Mean fluorescence intensity


Lactate dehydrogenase


Fluorescein isothiocyanate


Propidium iodide


Relative luminescence units


4′,6-Diamidino-2-phenylindole dihydrochloride


Green fluorescence protein


Endoplasmic reticulum


ER degradation enhancing α-mannosidase like protein 1


UDP-glucose glycoprotein glucosyltransferase 1


SEC62 homolog/preprotein translocation factor


Valosin-containing protein


Glucose-regulated protein 78/binding immunoglobulin protein


Inositol requiring kinase-1α


X-box binding protein 1


Protein kinase-like endoplasmic reticulum kinase


Eukariotic initiation factor-2α


Activating transcription factor 4


Activating transcription factor 6


CCAAT-enhancer-binding protein-β


C/EBP homologous protein/growth arrest and DNA damage 153


Tribbles homolog 3


p53 up-regulated modulator of apoptosis


TATA box binding protein antibodies




Relative fluorescence units


Unfolded protein response


Endoplasmic reticulum-associated protein degradation


Endoplasmic reticulum quality control



The work was supported by Italian Association for Cancer Research (IG15232 to CR); Italian Ministry of University and Research (RBFR12SOQ1 to C.R.); Istituto Ortopedico Rizzoli I.R.C.C.S. (5 x mille contributions to the Rizzoli Institute). We are grateful to Dr. Maria Alessandra Contino, Department of Pharmacy, University of Bari “Aldo Moro”, Bari, Italy, for the fruitful discussion, to Dr. Maria Pia Patrizio, Istituto Ortopedico Rizzoli I.R.C.C.S., for the help for the IC50 calculations and to Mr. Costanzo Costamagna, Department of Oncology, University of Torino, for the technical assistance.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

18_2018_2967_MOESM1_ESM.tif (7.4 mb)
Supplementary material 1 (TIFF 7569 kb) Supplementary Fig. 1 (Online Resource 1) Representative dot plots of apoptotic/necrotic cells. Doxorubicin-sensitive U-2OS and Saos-2 human osteosarcoma cell lines and their resistant variants (DX30, DX100 and DX580) were incubated 24 h with drug-free medium, 5 µM doxorubicin (dox) or H2S-releasing doxorubicin (Sdox), then the percentage of cells positively stained for Annexin V-FITC or propidium iodide (PI) was measured by flow cytometry in duplicates. Dot plots are representative of 1 out of 3 experiments
18_2018_2967_MOESM2_ESM.docx (17 kb)
Supplementary material 2 (DOCX 17 kb)
18_2018_2967_MOESM3_ESM.tif (1.9 mb)
Supplementary material 3 (TIFF 1895 kb) Supplementary Fig. 2 (Online Resource 3) Dose–response effect of doxorubicin and H2S-releasing doxorubicin on cell viability. Human doxorubicin-sensitive U-2OS cells and Saos-2 cells and their resistant sublines DX580 were incubated for 72 h with increasing concentrations (from 10 nM to 0.5 mM) of doxorubicin (dox) or H2S-releasing doxorubicin (Sdox). Cell viability measured by a chemiluminescence-based assay in quadruplicates. Data are mean ± SD (n = 6 independent experiments)
18_2018_2967_MOESM4_ESM.tif (1.2 mb)
Supplementary material 4 (TIFF 1229 kb) Supplementary Fig. 3 (Online Resource 4). Effects of doxorubicin and H2S-releasing doxorubicin on non-transformed cells. Human osteoblasts, human fibroblasts and rat H9c2 cardiomyocytes were grown in drug-free medium (ctrl) or in medium containing 5 µM doxorubicin (dox) or H2S-releasing doxorubicin (Sdox) for 24 h (panels a, c, e) or 72 h (panels b, d, f). a, c, e Extracellular release of LDH measured spectrophotometrically in triplicates. Data are mean ± SD (n = 3 independent experiments). *p < 0.001 for treated vs. untreated cells; °p < 0.001 for Sdox vs. dox. b, d, f Cell viability measured with a chemiluminescence-based method in quadruplicates. Data are mean ± SD (n = 3 independent experiments). *p < 0.002 for treated vs. untreated cells; °p < 0.05 for Sdox vs. dox
18_2018_2967_MOESM5_ESM.tif (1.5 mb)
Supplementary material 5 (TIFF 1530 kb) Supplementary Fig. 4 (Online Resource 5) Effects of verapamil on H2S-releasing doxorubicin accumulation and cytotoxicity. Doxorubicin-sensitive U-2OS human osteosarcoma cell line and its U-2OS/DX580 resistant variant were cultured for 6 h (panels a-b), 24 h (panels c-d) or 72 h (panels e–f) in drug-free medium (ctrl) or in medium containing 5 µM doxorubicin (dox) or H2S-releasing doxorubicin (Sdox), in the absence (-) or presence (+) of 50 µM verapamil (ver). a-b. Intracellular drug accumulation, measured by flow cytometry in duplicates, and expressed as mean fluorescence intensity (MFI). Data are mean ± SD (n = 6 independent experiments). *p < 0.001 for treated vs. untreated cells; °p < 0.001 for verapamil-treated vs. verapamil-untreated cells. c-d Extracellular release of LDH measured spectrophotometrically in triplicates. Data are mean ± SD (n = 4 independent experiments). *p < 0.001 for treated vs. untreated cells; °p < 0.01 for verapamil-treated vs. verapamil-untreated cells. e–f Cell viability measured with a chemiluminescence-based method in quadruplicates. Data are mean ± SD (n = 3 independent experiments). *p < 0.001 for treated vs. untreated cells; °p < 0.001 for verapamil-treated vs. verapamil-untreated cells
18_2018_2967_MOESM6_ESM.tif (4.5 mb)
Supplementary material 6 (TIFF 4630 kb) Supplementary Fig. 5 (Online Resource 6) Early intracellular localization of doxorubicin within sensitive osteosarcoma cells. U-2OS cells were incubated for 24 h with the GFP-KDEL-calreticulin expression vector to label endoplasmic reticulum (ER), then treated with 5 µM doxorubicin (dox) for 10 min, 20 min, 30 min, 1 h, 3 h, 6 h, 24 h. The intracellular localization of the drug was analyzed by fluorescence microscopy. Magnification: 63 × objective lens (1.42 numerical aperture); 10 × ocular lens Bar: 7.5 μm. The micrographs are representative of the dox localization after 20 min, corresponding to the time point with the highest accumulation within the ER, and are representative of 3 experiments with similar results
18_2018_2967_MOESM7_ESM.docx (32 kb)
Supplementary material 7 (DOCX 31 kb)
18_2018_2967_MOESM8_ESM.tif (1.2 mb)
Supplementary material 8 (TIFF 1184 kb) Supplementary Fig. 6 (Online Resource 8) Expression of ERAD/ERQC and UPR-related genes in doxorubicin-sensitive and doxorubicin-resistant osteosarcoma cells. a-c Hitmap of unfolded protein response (UPR)-related genes, cell death/survival related genes, ER-associated degradation/endoplasmic reticulum quality control (ERAD/ERQC)-related genes in U-2OS/DX30, U-2OS/DX100 and U-2OS/DX580 cells. The figure reports genes up-or down-regulated at least twofold, in at least one cell line, compared to untreated U-2OS cells (n = 6 independent experiments). The expression of each gene in U-2OS cells was considered 1 (not shown). The whole list of genes analyzed is reported in Supplementary Table 2 (Online Resource 7)
18_2018_2967_MOESM9_ESM.docx (28 kb)
Supplementary material 9 (DOCX 28 kb)
18_2018_2967_MOESM10_ESM.docx (29 kb)
Supplementary material 10 (DOCX 29 kb)
18_2018_2967_MOESM11_ESM.tif (338 kb)
Supplementary material 11 (TIFF 338 kb) Supplementary Fig. 7 (Online Resource 11) Expression of genes related to cell death and survival in doxorubicin-sensitive and doxorubicin-resistant osteosarcoma cells. Hitmap of genes related to cell death/survival in U-2OS and U-2OS/DX580 cells, after 24 h treatment with drug-free medium, 5 µM doxorubicin (dox) or H2S-releasing doxorubicin (Sdox). The figure reports genes up-or down-regulated at least twofold, in at least one cell line, compared to untreated U-2OS cells (n = 6 independent experiments). The expression of each gene in U-2OS cells was considered 1 (not shown). The whole list of genes analyzed is reported in Supplementary Tables 3-4 (Online Resources 9-10)


  1. 1.
    Hattinger CM, Fanelli M, Tavanti E, Vella S, Ferrari S, Picci P, Serra M (2015) Advances in emerging drugs for osteosarcoma. Expert Opin Emerg Drugs 20:495–514. CrossRefGoogle Scholar
  2. 2.
    Hattinger CM, Fanelli M, Tavanti E, Vella S, Riganti C, Picci P, Serra M (2017) Doxorubicin-resistant osteosarcoma: novel therapeutic approaches in sight? Future Oncol. 13:673–677. CrossRefGoogle Scholar
  3. 3.
    Gottesman MM, Fojo T, Bates SE (2002) Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2:48–58. CrossRefGoogle Scholar
  4. 4.
    Fanelli M, Hattinger CM, Vella S, Tavanti E, Michelacci F, Gudeman B, Barnett D, Picci P, Serra M (2016) Targeting ABCB1 and ABCC1 with their specific inhibitor CBT-1® can overcome drug resistance in osteosarcoma. Curr Cancer Drug Targets 16:261–274. CrossRefGoogle Scholar
  5. 5.
    Baldini N, Scotlandi K, Barbanti-Bròdano G et al (1995) Expression of P-glycoprotein in high-grade osteosarcomas in relation to clinical outcome. N Engl J Med 333:1380–1385. CrossRefGoogle Scholar
  6. 6.
    Serra M, Scotlandi K, Reverter-Branchat G, Ferrari S, Manara MC, Benini S, Incaprera M, Bertoni F, Mercuri M, Briccoli A, Bacci G, Picci P (2003) Value of P-glycoprotein and clinicopathologic factors as the basis for new treatment strategies in high-grade osteosarcoma of the extremities. J Clin Oncol 21:536–542. CrossRefGoogle Scholar
  7. 7.
    Serra M, Pasello M, Manara MC, Scotlandi K, Ferrari S, Bertoni F, Mercuri M, Alvegard TA, Picci P, Bacci G, Smeland S (2006) May P-glycoprotein status be used to stratify high-grade osteosarcoma patients? Results from the Italian/Scandinavian Sarcoma Group 1 treatment protocol. Int J Oncol 29:1459–1468. Google Scholar
  8. 8.
    Lipshultz SE, Karnik R, Sambatakos P, Franco VI, Ross SW, Miller TL (2014) Anthracycline-related cardiotoxicity in childhood cancer survivors. Curr Opin Cardiol 29:103–112. CrossRefGoogle Scholar
  9. 9.
    Callaghan R, Luk F, Bebawy M (2014) Inhibition of the multidrug resistance P-glycoprotein: time for a change of strategy? Drug Metab Dispos 42:623–631. CrossRefGoogle Scholar
  10. 10.
    Szakács G, Hall MD, Gottesman MM, Boumendjel A, Kachadourian R, Day BJ, Baubichon-Cortay H, Di Pietro A (2014) Targeting the Achilles heel of multidrug-resistant cancer by exploiting the fitness cost of resistance. Chem Rev 114:5753–5774. CrossRefGoogle Scholar
  11. 11.
    Tsouris V, Joo MK, Kim SH, Kwon IC, Won YY (2014) Nano carriers that enable co-delivery of chemotherapy and RNAi agents for treatment of drug-resistant cancers. Biotechnol Adv 32:1037–1050. CrossRefGoogle Scholar
  12. 12.
    Doublier S, Belisario DC, Polimeni M, Annaratone L, Riganti C, Allia E, Ghigo D, Bosia A, Sapino A (2012) HIF-1 activation induces doxorubicin resistance in MCF7 3-D spheroids via P-glycoprotein expression: a potential model of the chemo-resistance of invasive micropapillary carcinoma of the breast. BMC Cancer 12:e4. CrossRefGoogle Scholar
  13. 13.
    Roncuzzi L, Pancotti F, Baldini N (2014) Involvement of HIF-1α activation in the doxorubicin resistance of human osteosarcoma cells. Oncol Rep 32:389–394. CrossRefGoogle Scholar
  14. 14.
    Kopecka J, Rankin GM, Salaroglio IC, Poulsen SA, Riganti C (2016) P-glycoprotein-mediated chemoresistance is reversed by carbonic anhydrase XII inhibitors. Oncotarget 7:85861–85875. Google Scholar
  15. 15.
    Guo R, Lin J, Xu W, Shen N, Mo L, Zhang C, Feng J (2013) Hydrogen sulfide attenuates doxorubicin-induced cardiotoxicity by inhibition of the p38 MAPK pathway in H9c2 cells. Int J Mol Med 31:644–650. CrossRefGoogle Scholar
  16. 16.
    Sen S, Kawahara B, Gupta D, Tsai R, Khachatryan M, Roy-Chowdhuri S, Bose S, Yoon A, Faull K, Farias-Eisner R, Chaudhuri G (2015) Role of cystathionine β-synthase in human breast cancer. Free Radic Biol Med 86:228–238. CrossRefGoogle Scholar
  17. 17.
    Chegaev K, Rolando B, Cortese D, Gazzano E, Buondonno I, Lazzarato L, Fanelli M, Hattinger CM, Serra M, Riganti C, Fruttero R, Ghigo D, Gasco A (2016) H2S-donating doxorubicins may overcome cardiotoxicity and multidrug resistance. J Med Chem 59(10):4881–4889. CrossRefGoogle Scholar
  18. 18.
    Szabo C, Coletta C, Chao C, Módis K, Szczesny B, Papapetropoulos A, Hellmich MR (2013) Tumor-derived hydrogen sulfide, produced by cystathionine-β-synthase, stimulates bioenergetics, cell proliferation, and angiogenesis in colon cancer. Proc Natl Acad Sci USA 110:12474–12479. CrossRefGoogle Scholar
  19. 19.
    Hellmich MR, Coletta C, Chao C, Szabo C (2015) The therapeutic potential of cystathionine β-synthetase/hydrogen sulfide inhibition in cancer. Antioxid Redox Signal 22:424–448. CrossRefGoogle Scholar
  20. 20.
    Ma K, Liu Y, Zhu Q, Liu CH, Duan JL, Tan BK, Zhu YZ (2011) H2S donor, S-propargylcysteine, increases CSE in SGC-7901 and cancer-induced mice: evidence for a novel anti-cancer effect of endogenous H2S? PLoS One 6:e20525. CrossRefGoogle Scholar
  21. 21.
    Lu S, Gao Y, Huang X, Wang X (2014) GYY4137, a hydrogen sulfide (H2S) donor, shows potent anti-hepatocellular carcinoma activity through blocking the STAT3 pathway. Int J Oncol 44:1259–1267. CrossRefGoogle Scholar
  22. 22.
    Lv M, Li Y, Ji MH, Zhuang M, Tang JH (2014) Inhibition of invasion and epithelial-mesenchymal transition of human breast cancer cells by hydrogen sulfide through decreased phospho-p38 expression. Mol Med Rep 10:341–346. CrossRefGoogle Scholar
  23. 23.
    Kashfi K, Olson KR (2013) Biology and therapeutic potential of hydrogen sulfide and hydrogen sulfide-releasing chimeras. Biochem Pharmacol 85:689–703. CrossRefGoogle Scholar
  24. 24.
    Granados-Principal S, Quiles JL, Ramirez-Tortosa CL, Sanchez-Rovira P, Ramirez-Tortosa MC (2010) New advances in molecular mechanisms and the prevention of adriamycin toxicity by antioxidant nutrients. Food Chem Toxicol 48:1425–1438. CrossRefGoogle Scholar
  25. 25.
    Serra M, Scotlandi K, Manara MC, Maurici D, Lollini PL, De Giovanni C, Toffoli G, Baldini N (1993) Establishment and characterization of multidrug-resistant human osteosarcoma cell lines. Anticancer Res 13(2):323–329Google Scholar
  26. 26.
    Buondonno I, Gazzano E, Jean SR, Audrito V, Kopecka J, Fanelli M, Salaroglio IC, Costamagna C, Roato I, Mungo E, Hattinger CM, Deaglio S, Kelley SO, Serra M, Riganti C (2016) Mitochondria-targeted doxorubicin: a new therapeutic strategy against doxorubicin-resistant osteosarcoma. Mol Cancer Ther 15:2640–2652. CrossRefGoogle Scholar
  27. 27.
    Riganti C, Miraglia E, Viarisio D, Costamagna C, Pescarmona G, Ghigo D, Bosia A (2005) Nitric oxide reverts the resistance to doxorubicin in human colon cancer cells by inhibiting the drug efflux. Cancer Res 65:516–525Google Scholar
  28. 28.
    Ikeda M, Kurose A, Takatori E, Sugiyama T, Traganos F, Darzynkiewicz Z, Sawai T (2010) DNA damage detected with gammaH2AX in endometrioid adenocarcinoma cell lines. Int J Oncol 36:1081–1088Google Scholar
  29. 29.
    Sen N, Paul BD, Gadalla MM, Mustafa AK, Sen T, Xu R, Kim S, Snyder SH (2012) Hydrogen sulfide-linked sulfhydration of NF-κB mediates its antiapoptotic actions. Mol Cell 45:13–24. CrossRefGoogle Scholar
  30. 30.
    Kim I, Xu W, Reed J (2008) Cell death and endoplasmic reticulum stress: disease relevance and therapeutic opportunities. Nat Rev Drug Discov 7:1013–1030. CrossRefGoogle Scholar
  31. 31.
    Chevet E, Hetz C, Samali A (2015) Endoplasmic reticulum stress-activated cell reprogramming in oncogenesis. Cancer Discov 5:586–597. CrossRefGoogle Scholar
  32. 32.
    Li L, Rose P, Moore PK (2011) Hydrogen sulfide and cell signaling. Annu Rev Pharmacol Toxicol 51:169–187. CrossRefGoogle Scholar
  33. 33.
    Pan L, Aller SG (2015) Equilibrated atomic models of outward-facing P-glycoprotein and effect of ATP binding on structural dynamics. Sci Rep 5:e7880. CrossRefGoogle Scholar
  34. 34.
    Chiribau C, Gaccioli F, Huang C, Yuan C, Hatzoglou M (2010) Molecular symbiosis of chop and c/ebp beta isoform lip contributes to endoplasmic reticulum stress-induced apoptosis. Mol Cell Biol 30:3722–3731. CrossRefGoogle Scholar
  35. 35.
    Riganti C, Kopecka J, Panada E, Barak S, Rubinstein M (2015) The role of C/EBP-β LIP in multidrug resistance. J Natl Cancer Inst 107(5):djv046. CrossRefGoogle Scholar
  36. 36.
    Li T, Su L, Zhong N, Hao X, Zhong D, Singhal S, Liu X (2013) Salinomycin induces cell death with autophagy through activation of endoplasmic reticulum stress in human cancer cells. Autophagy 9:1057–1068. CrossRefGoogle Scholar
  37. 37.
    Cazanave SC, Elmi NA, Akazawa Y, Bronk SF, Mott JL, Gores GJ (2010) CHOP and AP-1 cooperatively mediate PUMA expression during lipoapoptosis. Am J Physiol Gastrointest Liver Physiol 299:G236–G243. CrossRefGoogle Scholar
  38. 38.
    Li J, Lee B, Lee AS (2006) Endoplasmic reticulum stress-induced apoptosis: multiple pathways and activation of p53-up-regulated modulator of apoptosis (PUMA) and NOXA by p53. J Biol Chem 281:7260–7270. CrossRefGoogle Scholar
  39. 39.
    Maraldi NM, Zini N, Santi S, Scotlandi K, Serra M, Baldini N (1999) P-glycoprotein subcellular localization and cell morphotype in MDR1 gene-transfected human osteosarcoma cells. Biol Cell 91:17–28CrossRefGoogle Scholar
  40. 40.
    Simunek T, Sterba M, Popelova O, Adamcova M, Hrdina R, Gersl V (2009) Anthracycline-induced cardiotoxicity: overview of studies examining the roles of oxidative stress and free cellular iron. Pharmacol Rep 61:154–171CrossRefGoogle Scholar
  41. 41.
    Bruce King S (2013) Potential biological chemistry of hydrogen sulfide (H2S) with the nitrogen oxides. Free Radic Biol Med 55:1–7. CrossRefGoogle Scholar
  42. 42.
    Tesei A, Brigliadori G, Carloni S (2012) Organosulfur derivatives of the HDAC inhibitor valproic acid sensitize human lung cancer cell lines to apoptosis and to cisplatin cytotoxicity. J Cell Physiol 227:3389–3396. CrossRefGoogle Scholar
  43. 43.
    Chamberlain GR, Tulumello DV, Kelley SO (2013) Targeted delivery of doxorubicin to mitochondria. ACS Chem Biol 8:1389–1395. CrossRefGoogle Scholar
  44. 44.
    Salaroglio IC, Panada E, Moiso E, Buondonno I, Provero P, Rubinstein M, Kopecka J, Riganti C (2017) PERK induces resistance to cell death elicited by endoplasmic reticulum stress and chemotherapy. Mol Cancer 16:e91. CrossRefGoogle Scholar
  45. 45.
    Meir O, Dvash E, Werman A, Rubinstein M (2010) C/ebp-beta regulates endoplasmic reticulum stress-triggered cell death in mouse and human models. PLoS One 5:e9516. CrossRefGoogle Scholar
  46. 46.
    Hetz C (2012) The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nat Rev Mol Cell Biol 13:89–102. CrossRefGoogle Scholar
  47. 47.
    Bastola P, Neums L, Schoenen FJ, Chien J (2016) VCP inhibitors induce endoplasmic reticulum stress, cause cell cycle arrest, trigger caspase-mediated cell death and synergistically kill ovarian cancer cells in combination with Salubrinal. Mol Oncol 10:1559–1574. CrossRefGoogle Scholar
  48. 48.
    Panaretakis T, Kepp O, Brockmeier U, Tesniere A, Bjorklund AC, Chapman DC, Durchschlag M, Joza N, Pierron G, van Endert P, Yuan J, Zitvogel L, Madeo F, Williams DB, Kroemer G (2009) Mechanisms of pre-apoptotic calreticulin exposure in immunogenic cell death. Embo J 28:578–590. CrossRefGoogle Scholar
  49. 49.
    Swartz DJ, Mok L, Botta SK, Singh A, Altenberg GA, Urbatsch IL (2014) Directed evolution of P-glycoprotein cysteines reveals site-specific, non-conservative substitutions that preserve multidrug resistance. Biosci Rep 34:e00116. CrossRefGoogle Scholar
  50. 50.
    Galluzzi L, Buqué A, Kepp O, Zitvogel L, Kroemer G (2017) Immunogenic cell death in cancer and infectious disease. Nat Rev Immunol 17:97–111. CrossRefGoogle Scholar
  51. 51.
    Krishnan N, Fu C, Pappin DJ, Tonks NK (2011) H2S-Induced sulfhydration of the phosphatase PTP1B and its role in the endoplasmic reticulum stress response. Sci Signal 4:ra86. CrossRefGoogle Scholar
  52. 52.
    Gao XH, Krokowski D, Guan BJ, Bederman I, Majumder M, Parisien M, Diatchenko L, Kabil O, Willard B, Banerjee R, Wang B, Bebek G, Evans CR, Fox PL, Gerson SL, Hoppel CL, Liu M, Arvan P, Hatzoglou M (2015) Quantitative H2S-mediated protein sulfhydration reveals metabolic reprogramming during the integrated stress response. Elife 4:e10067. CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  • Ilaria Buondonno
    • 1
  • Elena Gazzano
    • 1
  • Elisa Tavanti
    • 2
  • Konstantin Chegaev
    • 3
  • Joanna Kopecka
    • 1
  • Marilù Fanelli
    • 2
  • Barbara Rolando
    • 3
  • Roberta Fruttero
    • 3
  • Alberto Gasco
    • 3
  • Claudia Hattinger
    • 2
  • Massimo Serra
    • 2
  • Chiara Riganti
    • 1
    Email author
  1. 1.Department of OncologyUniversity of TorinoTorinoItaly
  2. 2.Laboratory of Experimental Oncology, Pharmacogenomics and Pharmacogenetics Research UnitOrthopaedic Rizzoli Institute I.R.C.C.SBolognaItaly
  3. 3.Department of Drug Science and TechnologyUniversity of TorinoTorinoItaly

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