Abstract
Pancreatic ductal adenocarcinoma is a devastating cancer and is the fourth-leading cause of cancer death in the USA. Zinc is abundant in the pancreas, but its role in pancreatic cancer remains elusive. The aim of this study is to determine effects of zinc chelators in pancreatic cancer. Pdx1Cre and LSL-KrasG12D mice expressing an oncogenic mutation of KRAS develop pancreatic intraepithelial neoplasia in the pancreas. We found that EPCAM + tumors developed in the mouse pancreas store zinc that is detectable by fluorescence-activated cell sorting using N-(6-methoxy-8-quinolyl)-p-toluenesulfonamide (TSQ), a fluorescence chelator. EPCAM + TSQ + tumor cells isolated from the mouse pancreas formed organoids in matrigel. Upon treatment with N,N,N′,N′-tetrakis(2-pyridinylmethyl)-1,2-ethanediamine (TPEN), a zinc chelator, the organoids degenerated and its negative effect was rescued by co-treatment with zinc, indicating that zinc is necessary for the growth and survival of tumor organoids. Different from TPEN, TSQ treatment did not affect the organoid growth and survival. Interestingly, co-treatment with TSQ and zinc resulted in strong emission of TSQ fluorescence in the organoid and its degeneration. The combination of zinc with TSQ, but not with TPEN, also induced cell death in PANC-1, a human pancreatic cancer cell line. These results suggest that a TSQ-zinc complex formed in pancreatic tumors induces cell death if zinc is overloaded.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Hezel AF, Kimmelman AC, Stanger BZ, Bardeesy N, Depinho RA (2006) Genetics and biology of pancreatic ductal adenocarcinoma. Genes Dev 20:1218–1249. https://doi.org/10.1101/gad.1415606
Liou GY, Doppler H, Necela B, Krishna M, Crawford HC, Raimondo M, Storz P (2013) Macrophage-secreted cytokines drive pancreatic acinar-to-ductal metaplasia through NF-kappaB and MMPs. J Cell Biol 202:563–577. https://doi.org/10.1083/jcb.201301001
Kopp JL, von Figura G, Mayes E, Liu FF, Dubois CL, Morris JP, Pan FC, Akiyama H, Wright CV, Jensen K, Hebrok M, Sander M (2012) Identification of Sox9-dependent acinar-to-ductal reprogramming as the principal mechanism for initiation of pancreatic ductal adenocarcinoma. Cancer Cell 22:737–750. https://doi.org/10.1016/j.ccr.2012.10.025
Asahina K, Balog S, Hwang E, Moon E, Wan E, Skrypek K, Chen Y, Fernandez J, Romo J, Yang Q, Lai K, French SW, Tsukamoto H (2020) Moderate alcohol intake promotes pancreatic ductal adenocarcinoma development in mice expressing oncogenic Kras. Am J Physiol Gastrointest Liver Physiol 318:G265-276. https://doi.org/10.1152/ajpgi.00218.2019
Morris JPt, Wang SC, Hebrok M, (2010) KRAS, Hedgehog, Wnt and the twisted developmental biology of pancreatic ductal adenocarcinoma. Nat Rev Cancer 10:683–695. https://doi.org/10.1038/nrc2899
Chiaravalli M, Reni M, O’Reilly EM (2017) Pancreatic ductal adenocarcinoma: state-of-the-art 2017 and new therapeutic strategies. Cancer Treat Rev 60:32–43. https://doi.org/10.1016/j.ctrv.2017.08.007
Siegel RL, Miller KD, Jemal A (2018) Cancer statistics, 2018. CA Cancer J Clin 68:7–30. https://doi.org/10.3322/caac.21442
Martin AM, Hidalgo M, Alvarez R, Arrazubi V, Martinez-Galan J, Salgado M, Macarulla T, Carrato A (2018) From first line to sequential treatment in the management of metastatic pancreatic cancer. J Cancer 9:1978–1988. https://doi.org/10.7150/jca.23716
Kunk PR, Bauer TW, Slingluff CL, Rahma OE (2016) From bench to bedside a comprehensive review of pancreatic cancer immunotherapy. J Immunother Cancer 4:14. https://doi.org/10.1186/s40425-016-0119-z
Kleeff J, Korc M, Apte M, La Vecchia C, Johnson CD, Biankin AV, Neale RE, Tempero M, Tuveson DA, Hruban RH, Neoptolemos JP (2016) Pancreatic cancer. Nat Rev Dis Primers 2:16022. https://doi.org/10.1038/nrdp.2016.22
Kanda M, Matthaei H, Wu J, Hong SM, Yu J, Borges M, Hruban RH, Maitra A, Kinzler K, Vogelstein B, Goggins M (2012) Presence of somatic mutations in most early-stage pancreatic intraepithelial neoplasia. Gastroenterology 142:730–733. https://doi.org/10.1053/j.gastro.2011.12.042
Hingorani SR, Petricoin EF, Maitra A, Rajapakse V, King C, Jacobetz MA, Ross S, Conrads TP, Veenstra TD, Hitt BA, Kawaguchi Y, Johann D, Liotta LA, Crawford HC, Putt ME, Jacks T, Wright CV, Hruban RH, Lowy AM, Tuveson DA (2003) Preinvasive and invasive ductal pancreatic cancer and its early detection in the mouse. Cancer Cell 4:437–450
Sekler I, Sensi SL, Hershfinkel M, Silverman WF (2007) Mechanism and regulation of cellular zinc transport. Mol Med 13:337–343. https://doi.org/10.2119/2007-00037.Sekler
Kelleher SL, McCormick NH, Velasquez V, Lopez V (2011) Zinc in specialized secretory tissues: roles in the pancreas, prostate, and mammary gland. Adv Nutr 2:101–111. https://doi.org/10.3945/an.110.000232
Andrews JC, Nolan JP, Hammerstedt RH, Bavister BD (1995) Characterization of N-(6-methoxy-8-quinolyl)-p-toluenesulfonamide for the detection of zinc in living sperm cells. Cytometry 21:153–159. https://doi.org/10.1002/cyto.990210207
Meeusen JW, Tomasiewicz H, Nowakowski A, Petering DH (2011) TSQ (6-methoxy-8-p-toluenesulfonamido-quinoline), a common fluorescent sensor for cellular zinc, images zinc proteins. Inorg Chem 50:7563–7573. https://doi.org/10.1021/ic200478q
Baltaci AK, Mogulkoc R, Baltaci SB (2019) Review: the role of zinc in the endocrine system. Pak J Pharm Sci 32:231–239
Jayaraman AK, Jayaraman S (2011) Increased level of exogenous zinc induces cytotoxicity and up-regulates the expression of the ZnT-1 zinc transporter gene in pancreatic cancer cells. J Nutr Biochem 22:79–88. https://doi.org/10.1016/j.jnutbio.2009.12.001
Donadelli M, DallaPozza E, Costanzo C, Scupoli MT, Scarpa A, Palmieri M (2008) Zinc depletion efficiently inhibits pancreatic cancer cell growth by increasing the ratio of antiproliferative/proliferative genes. J Cell Biochem 104:202–212. https://doi.org/10.1002/jcb.21613
Li M, Zhang Y, Liu Z, Bharadwaj U, Wang H, Wang X, Zhang S, Liuzzi JP, Chang SM, Cousins RJ, Fisher WE, Brunicardi FC, Logsdon CD, Chen C, Yao Q (2007) Aberrant expression of zinc transporter ZIP4 (SLC39A4) significantly contributes to human pancreatic cancer pathogenesis and progression. Proc Natl Acad Sci U S A 104:18636–18641. https://doi.org/10.1073/pnas.0709307104
Frederickson CJ, Kasarskis EJ, Ringo D, Frederickson RE (1987) A quinoline fluorescence method for visualizing and assaying the histochemically reactive zinc (bouton zinc) in the brain. J Neurosci Methods 20:91–103. https://doi.org/10.1016/0165-0270(87)90042-2
Nitzan YB, Sekler I, Silverman WF (2004) Histochemical and histofluorescence tracing of chelatable zinc in the developing mouse. J Histochem Cytochem 52:529–539. https://doi.org/10.1177/002215540405200411
Horton TM, Allegretti PA, Lee S, Moeller HP, Smith M, Annes JP (2019) Zinc-chelating small molecules preferentially accumulate and function within pancreatic β cells. Cell Chem Biol 26:213–222. https://doi.org/10.1016/j.chembiol.2018.10.019
Boj SF, Hwang CI, Baker LA, Chio II, Engle DD, Corbo V, Jager M, Ponz-Sarvise M, Tiriac H, Spector MS, Gracanin A, Oni T, Yu KH, van Boxtel R, Huch M, Rivera KD, Wilson JP, Feigin ME, Ohlund D, Handly-Santana A, Ardito-Abraham CM, Ludwig M, Elyada E, Alagesan B, Biffi G, Yordanov GN, Delcuze B, Creighton B, Wright K, Park Y, Morsink FH, Molenaar IQ, BorelRinkes IH, Cuppen E, Hao Y, Jin Y, Nijman IJ, Iacobuzio-Donahue C, Leach SD, Pappin DJ, Hammell M, Klimstra DS, Basturk O, Hruban RH, Offerhaus GJ, Vries RG, Clevers H, Tuveson DA (2015) Organoid models of human and mouse ductal pancreatic cancer. Cell 160:324–338. https://doi.org/10.1016/j.cell.2014.12.021
Skrypek K, Balog S, Eriguchi Y, Asahina K (2021) Inhibition of stearoyl-CoA desaturase induces the unfolded protein response in pancreatic tumors and suppresses their growth. Pancreas 50:219–226. https://doi.org/10.1097/mpa.0000000000001737
Huch M, Bonfanti P, Boj SF, Sato T, Loomans CJ, van de Wetering M, Sojoodi M, Li VS, Schuijers J, Gracanin A, Ringnalda F, Begthel H, Hamer K, Mulder J, van Es JH, de Koning E, Vries RG, Heimberg H, Clevers H (2013) Unlimited in vitro expansion of adult bi-potent pancreas progenitors through the Lgr5/R-spondin axis. EMBO J 32:2708–2721. https://doi.org/10.1038/emboj.2013.204
Deer EL, Gonzalez-Hernandez J, Coursen JD, Shea JE, Ngatia J, Scaife CL, Firpo MA, Mulvihill SJ (2010) Phenotype and genotype of pancreatic cancer cell lines. Pancreas 39:425–435. https://doi.org/10.1097/MPA.0b013e3181c15963
Zhang Y, Yang J, Cui X, Chen Y, Zhu VF, Hagan JP, Wang H, Yu X, Hodges SE, Fang J, Chiao PJ, Logsdon CD, Fisher WE, Brunicardi FC, Chen C, Yao Q, Fernandez-Zapico ME, Li M (2013) A novel epigenetic CREB-miR-373 axis mediates ZIP4-induced pancreatic cancer growth. EMBO Mol Med 5:1322–1334. https://doi.org/10.1002/emmm.201302507
Liu M, Zhang Y, Yang J, Zhan H, Zhou Z, Jiang Y, Shi X, Fan X, Zhang J, Luo W, Fung KA, Xu C, Bronze MS, Houchen CW, Li M (2021) Zinc-dependent regulation of ZEB1 and YAP1 co-activation promotes epithelial-mesenchymal transition plasticity and metastasis in pancreatic cancer. Gastroenterology 160:1771–1783. https://doi.org/10.1053/j.gastro.2020.12.077
Rybarczyk P, Vanlaeys A, Brassart B, Dhennin-Duthille I, Chatelain D, Sevestre H, Ouadid-Ahidouch H, Gautier M (2017) The transient receptor potential melastatin 7 channel regulates pancreatic cancer cell invasion through the Hsp90α/uPA/MMP2 pathway. Neoplasia 19:288–300. https://doi.org/10.1016/j.neo.2017.01.004
Xie J, Cheng CS, Zhu XY, Shen YH, Song LB, Chen H, Chen Z, Liu LM, Meng ZQ (2019) Magnesium transporter protein solute carrier family 41 member 1 suppresses human pancreatic ductal adenocarcinoma through magnesium-dependent Akt/mTOR inhibition and bax-associated mitochondrial apoptosis. Aging 11:2681–2698. https://doi.org/10.18632/aging.101940
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This study was supported by the pilot project from the Southern California Research Center for ALPD & Cirrhosis (National Institutes of Health P50AA11999) and USC Dean’s pilot project program.
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Asahina, K. Induction of Cell Death in Pancreatic Tumors by Zinc and Its Fluorescence Chelator TSQ. Biol Trace Elem Res 200, 1667–1676 (2022). https://doi.org/10.1007/s12011-021-02770-7
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DOI: https://doi.org/10.1007/s12011-021-02770-7