Archives of Toxicology

, Volume 93, Issue 2, pp 533–546 | Cite as

sFRP1 has a biphasic effect on doxorubicin-induced cardiotoxicity in a cellular location-dependent manner in NRCMs and Rats

  • Yuehuai Hu
  • Zhen Guo
  • Jing LuEmail author
  • Panxia Wang
  • Shuya Sun
  • Yiqiang Zhang
  • Jingyan Li
  • Qiyao Zheng
  • Kaiteng Guo
  • Junjian Wang
  • Jianmin JiangEmail author
  • Peiqing LiuEmail author
Organ Toxicity and Mechanisms


Doxorubicin (Dox) is an effective anticancer drug, however, its clinical application is restricted by the life-threatening cardiotoxic effects. Secreted Frizzled-related protein 1 (sFRP1) has been reported to participate in both the cancer and cardiovascular diseases and was one of the differential expression genes in normal hearts compared with Dox-treated hearts. Thus, it is important to reveal the potential role of sFRP1 in Dox-induced cardiotoxicity. Here, we show that sFRP1 has a biphasic effect on Dox-induced cardiotoxicity in a location-dependent manner. The secretion of sFRP1 was significantly increased in Dox-treated neonatal rat cardiomyocytes (NRCMs) (1 µM) and SD rats (5 mg/kg/injection at day 1, 5, and 9, i.p.). Adding the anti-sFRP1 antibody (0.5 µg/ml) and inhibiting sFRP1 secretion by caffeine (5 mM) both relieved Dox-induced cardiotoxicity through activating Wnt/β-catenin signaling, whereas increasing the secretion of sFRP1 by heparin (100 µg/ml) had the opposite effect. The intracellular level of sFRP1 was significantly decreased after Dox treatment both in vitro and in vivo. Knockdown of sFRP1 by sgRNA aggravated Dox-induced cardiotoxicity, while moderate overexpression of sFRP1 by Ad-sFRP1 exhibited protective effect. Besides, poly(ADP-ribosyl) polymerase-1 (PARP1) was screened as an interacting partner of sFRP1 in NRCMs by mass spectrometry. Our results suggested that the intracellular sFRP1 protected NRCMs from Dox-induced cardiotoxicity by interacting with PARP1. Thus, our results provide a novel evidence that sFRP1 has a biphasic effect on Dox-induced cardiotoxicity. In addition, the oversecretion of sFRP1 might be used as a biomarker to indicate the occurrence of cardiotoxicity induced by Dox treatment.


sFRP1 Doxorubicin Cardiotoxicity Wnt/β-catenin PARP1 



This research was supported by Grants from the 111 Project (No. B16047), National Natural Science Foundation of China (30670837, 81473205, 81673433, 81803521, 81872860), the Natural Science Foundation of Guangdong Province, China (No. 2016A030313211), Indigenous Innovative Research Team of Guangdong Province (2017BT01Y093), National Major Special Projects for the Creation and Manufacture of New Drugs (2018ZX09301031-001), Major Project of Platform Construction Education Department of Guangdong Province (No. 2014GKPT002), and Special Program for Applied Science and Technology of Guangdong Province (No. 2015B020232009 and 2014B020210003), National Engineering and Technology Research Center for New drug Druggability Evaluation (Seed Program of Guangdong Province, 2017B090903004), Guangzhou Science and Technology Program Project (No. 201604020121), Medical Scientific Research Foundation of Guangdong Province (No. A2018078).

Compliance with ethical standards

Conflict of interest

All authors declare that there is no conflict of interest.

Ethics statement

The animal experimental procedures were passed by the Research Ethics Committee of Sun Yat-sen University, and were conducted in accordance with the Guide for the Care and Use of Laboratory Animals (NIH Publication No. 85-23, revised 1996).

Supplementary material

204_2018_2342_MOESM1_ESM.doc (11.5 mb)
Supplementary material 1 (DOC 11736 KB)


  1. Barandon L, Couffinhal T, Ezan J et al (2003) Reduction of infarct size and prevention of cardiac rupture in transgenic mice overexpressing FrzA. Circulation 108(18):2282–2289. CrossRefGoogle Scholar
  2. Barandon L, Dufourcq P, Costet P et al (2005) Involvement of FrzA/sFRP-1 and the Wnt/frizzled pathway in ischemic preconditioning. Circ Res 96(12):1299–1306. CrossRefGoogle Scholar
  3. Bartha E, Solti I, Szabo A et al (2011) Regulation of kinase cascade activation and heat shock protein expression by poly(ADP-ribose) polymerase inhibition in doxorubicin-induced heart failure. J Cardiovasc Pharmacol 58(4):380–391. CrossRefGoogle Scholar
  4. Berthiaume JM, Oliveira PJ, Fariss MW, Wallace KB (2005) Dietary vitamin E decreases doxorubicin-induced oxidative stress without preventing mitochondrial dysfunction. Cardiovasc Toxicol 5(3):257–267CrossRefGoogle Scholar
  5. Bharathi Priya L, Baskaran R, Huang CY, Vijaya Padma V (2017) Neferine modulates IGF-1R/Nrf2 signaling in doxorubicin treated H9c2 cardiomyoblasts. J Cell Biochemi 119(2):1441–1452. CrossRefGoogle Scholar
  6. Cardinale D, Colombo A, Sandri MT et al (2006) Prevention of high-dose chemotherapy-induced cardiotoxicity in high-risk patients by angiotensin-converting enzyme inhibition. Circulation 114(23):2474–2481. CrossRefGoogle Scholar
  7. Carvalho FS, Burgeiro A, Garcia R, Moreno AJ, Carvalho RA, Oliveira PJ (2014) Doxorubicin-induced cardiotoxicity: from bioenergetic failure and cell death to cardiomyopathy. Med Res Rev 34(1):106–135. CrossRefGoogle Scholar
  8. Chiarugi A, Moskowitz MA (2002) Cell biology. PARP-1—a perpetrator of apoptotic cell death? Science 297(5579):200–201. CrossRefGoogle Scholar
  9. Chung MT, Lai HC, Sytwu HK et al (2009) SFRP1 and SFRP2 suppress the transformation and invasion abilities of cervical cancer cells through Wnt signal pathway. Gynecol Oncol 112(3):646–653. CrossRefGoogle Scholar
  10. Coelho AR, Martins TR, Couto R et al (2017) Berberine-induced cardioprotection and Sirt3 modulation in doxorubicin-treated H9c2 cardiomyoblasts. Biochim Biophys Acta 1863(11):2904–2923. CrossRefGoogle Scholar
  11. Dahl E, Veeck J, An H et al (2005) Epigenetic inactivation of the WNT antagonist SFRP1 in breast cancer. Verh Dtsch Ges Pathol 89:169–177Google Scholar
  12. Efremova AS, Shram SI, Myasoedov NF (2015) Doxorubicin causes transient activation of protein poly(ADP-ribosyl)ation in H9c2 cardiomyocytes. Dokl Biochem Biophys 464:333–337. CrossRefGoogle Scholar
  13. Elzi DJ, Song M, Hakala K, Weintraub ST, Shiio Y (2012) Wnt antagonist SFRP1 functions as a secreted mediator of senescence. Mol Cell Biol 32(21):4388–4399. CrossRefGoogle Scholar
  14. Gao S, Li H, Feng XJ et al (2015) alpha-Enolase plays a catalytically independent role in doxorubicin-induced cardiomyocyte apoptosis and mitochondrial dysfunction. J Mol Cell Cardiol 79:92–103. CrossRefGoogle Scholar
  15. Ghoshal A, Ghosh SS (2015) Expression, purification, and therapeutic implications of recombinant sFRP1. Appl Biochem Biotechnol 175(4):2087–2103. CrossRefGoogle Scholar
  16. Hang P, Zhao J, Sun L et al (2017) Brain-derived neurotrophic factor attenuates doxorubicin-induced cardiac dysfunction through activating Akt signalling in rats. J Cell Mol Med 21(4):685–696. CrossRefGoogle Scholar
  17. Hosseini A, Bakhtiari E, Mousavi SH (2017) Protective effect of Hibiscus sabdariffa on doxorubicin-induced cytotoxicity in H9c2 cardiomyoblast cells. Iran J Pharm Res IJPR 16(2):708–713Google Scholar
  18. Hullin R, Metrich M, Sarre A et al (2017) Diverging effects of enalapril or eplerenone in primary prevention against doxorubicin-induced cardiotoxicity. Cardiovasc Res 114(2):272–281. CrossRefGoogle Scholar
  19. Ichikawa Y, Ghanefar M, Bayeva M et al (2014) Cardiotoxicity of doxorubicin is mediated through mitochondrial iron accumulation. J Clin Investig 124(2):617–630. CrossRefGoogle Scholar
  20. Kalay N, Basar E, Ozdogru I et al (2006) Protective effects of carvedilol against anthracycline-induced cardiomyopathy. J Am Coll Cardiol 48(11):2258–2262. CrossRefGoogle Scholar
  21. Klopocki E, Kristiansen G, Wild PJ et al (2004) Loss of SFRP1 is associated with breast cancer progression and poor prognosis in early stage tumors. Int J Oncol 25(3):641–649Google Scholar
  22. Lebrecht D, Setzer B, Ketelsen UP, Haberstroh J, Walker UA (2003) Time-dependent and tissue-specific accumulation of mtDNA and respiratory chain defects in chronic doxorubicin cardiomyopathy. Circulation 108(19):2423–2429. CrossRefGoogle Scholar
  23. Li K, Sung RY, Huang WZ et al (2006a) Thrombopoietin protects against in vitro and in vivo cardiotoxicity induced by doxorubicin. Circulation 113(18):2211–2220. CrossRefGoogle Scholar
  24. Li L, Takemura G, Li Y et al (2006b) Preventive effect of erythropoietin on cardiac dysfunction in doxorubicin-induced cardiomyopathy. Circulation 113(4):535–543. CrossRefGoogle Scholar
  25. Liu J, Wang Y, Du W et al (2013) Wnt1 inhibits hydrogen peroxide-induced apoptosis in mouse cardiac stem cells. PLoS One 8(3):e58883. CrossRefGoogle Scholar
  26. Lou H, Danelisen I, Singal PK (2005) Involvement of mitogen-activated protein kinases in adriamycin-induced cardiomyopathy. Am J Physiol Heart Circ Physiol 288(4):H1925–H1930. CrossRefGoogle Scholar
  27. Moulin M, Piquereau J, Mateo P et al (2015) Sexual dimorphism of doxorubicin-mediated cardiotoxicity: potential role of energy metabolism remodeling. Circ Heart Fail 8(1):98–108. CrossRefGoogle Scholar
  28. Pacher P, Liaudet L, Bai P et al (2002) Activation of poly(ADP-ribose) polymerase contributes to development of doxorubicin-induced heart failure. J Pharmacol Exp Ther 300(3):862–867CrossRefGoogle Scholar
  29. Rajagopalan S, Politi PM, Sinha BK, Myers CE (1988) Adriamycin-induced free radical formation in the perfused rat heart: implications for cardiotoxicity. Cancer Res 48(17):4766–4769Google Scholar
  30. Shi R, Huang CC, Aronstam RS, Ercal N, Martin A, Huang YW (2009) N-acetylcysteine amide decreases oxidative stress but not cell death induced by doxorubicin in H9c2 cardiomyocytes. BMC Pharmacol 9:7. CrossRefGoogle Scholar
  31. Shi Y, Moon M, Dawood S, McManus B, Liu PP (2011) Mechanisms and management of doxorubicin cardiotoxicity. Herz 36(4):296–305. CrossRefGoogle Scholar
  32. Shulewitz M, Soloviev I, Wu T, Koeppen H, Polakis P, Sakanaka C (2006) Repressor roles for TCF-4 and Sfrp1 in Wnt signaling in breast cancer. Oncogene 25(31):4361–4369. CrossRefGoogle Scholar
  33. Sterba M, Popelova O, Vavrova A et al (2013) Oxidative stress, redox signaling, and metal chelation in anthracycline cardiotoxicity and pharmacological cardioprotection. Antioxid Redox Signal 18(8):899–929. CrossRefGoogle Scholar
  34. Tao J, Chen BD, Ma YT et al (2015) FrzA gene protects cardiomyocytes from H2O2-induced oxidative stress through restraining the Wnt/Frizzled pathway. Lipids Health Dis 14:90. CrossRefGoogle Scholar
  35. Tao J, Abudoukelimu M, Ma YT et al (2016) Secreted frizzled related protein 1 protects H9C2 cells from hypoxia/re-oxygenation injury by blocking the Wnt signaling pathway. Lipids Health Dis 15:72. CrossRefGoogle Scholar
  36. Tokarska-Schlattner M, Zaugg M, Zuppinger C, Wallimann T, Schlattner U (2006) New insights into doxorubicin-induced cardiotoxicity: the critical role of cellular energetics. J Mol Cell Cardiol 41(3):389–405. CrossRefGoogle Scholar
  37. Uren A, Reichsman F, Anest V et al (2000) Secreted frizzled-related protein-1 binds directly to Wingless and is a biphasic modulator of Wnt signaling. J Biol Chem 275(6):4374–4382CrossRefGoogle Scholar
  38. Vandenwijngaert S, Swinnen M, Walravens AS et al (2017) Decreased soluble guanylate cyclase contributes to cardiac dysfunction induced by chronic doxorubicin treatment in mice. Antioxid Redox Signal 26(4):153–164. CrossRefGoogle Scholar
  39. Wu F, Li J, Guo N, Wang XH, Liao YQ (2017) MiRNA-27a promotes the proliferation and invasion of human gastric cancer MGC803 cells by targeting SFRP1 via Wnt/beta-catenin signaling pathway. Am J Cancer Res 7(3):405–416Google Scholar
  40. Xavier CP, Melikova M, Chuman Y, Uren A, Baljinnyam B, Rubin JS (2014) Secreted Frizzled-related protein potentiation versus inhibition of Wnt3a/beta-catenin signaling. Cell Signal 26(1):94–101. CrossRefGoogle Scholar
  41. Zhang Y, Tu C, Zhang D et al (2015) Wnt/beta-catenin and Wnt5a/Ca pathways regulate proliferation and apoptosis of keratinocytes in psoriasis lesions. Cell Physiol Biochem 36(5):1890–1902 CrossRefGoogle Scholar
  42. Zhang Y, Wang Y, Ma Z et al (2017) Ginsenoside Rb1 inhibits doxorubicin-triggered H9C2 cell apoptosis via aryl hydrocarbon receptor. Biomol Ther 25(2):202–212. CrossRefGoogle Scholar
  43. Zhang N, Zhang Y, Zhao S, Sun Y (2018) Septin4 as a novel binding partner of PARP1 contributes to oxidative stress induced human umbilical vein endothelial cells injure. Biochem Biophys Res Commun 496(2):621–627. CrossRefGoogle Scholar
  44. Zhou SG, Zhou SF, Huang HQ, Chen JW, Huang M, Liu PQ (2006) Proteomic analysis of hypertrophied myocardial protein patterns in renovascularly hypertensive and spontaneously hypertensive rats. J Proteome Res 5(11):2901–2908. CrossRefGoogle Scholar
  45. Zu G, Guo J, Che N, Zhou T, Zhang X (2016) Protective effects of ginsenoside Rg1 on intestinal ischemia/reperfusion injury-induced oxidative stress and apoptosis via activation of the Wnt/beta-catenin pathway. Sci Rep 6:38480. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.School of Pharmaceutical Sciences, Guangzhou Higher Education Mega CenterSun Yat-sen UniversityGuangzhouPeople’s Republic of China
  2. 2.School of Pharmaceutical SciencesGuangzhou University of Chinese MedicineGuangzhouPeople’s Republic of China
  3. 3.Division of Cardiology, Department of Medicine, Center for Cardiovascular Biology, Institute for Stem Cell and Regenerative MedicineUniversity of WashingtonSeattleUSA

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