Skip to main content

Advertisement

SpringerLink
Log in

Effects of increased accumulation of doxorubicin due to emodin on efflux transporter and LRP1 expression in lung adenocarcinoma and colorectal carcinoma cells

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Treatment with doxorubicin (dox) and emodin, separately and together, under normoxic and hypoxia-like conditions induced by CoCl2, led to greater intracellular compound accumulation over 10 h post-addition in the presence of CoCl2 in lung adenocarcinoma (A549) and colorectal carcinoma (HCT-15) cell lines. Confocal microscopy revealed that emodin, by itself, showed high cytosolic distribution in both cell lines, at 40 min post-addition but had entered the nuclei by 2 h, while dox entered the nuclei by 40 min. Both compounds modulated the expression of the efflux transporters (PgP, ABCG2, or MRP1-4) and the endocytic receptor, low-density lipoprotein receptor-related protein 1 (LRP1), to different extents under the study conditions. Efflux transporter upregulation was linked to lower intracellular compound levels due to greater efflux. Increased dox accumulation was accompanied by unaltered expression or upregulation of LRP1 in A549 cells. In both cell lines, increased accumulation of dox and emodin was observed whenever LRP1 and the efflux transporters known to transport dox and emodin were all up- or downregulated concomitantly. Increased growth inhibition was linked to co-treatment with dox and emodin and with increased ligand accumulation. The results presented in this study raise the hypothesis that higher production of LRP1 protein may be associated with higher endocytosis of upregulated transporter proteins at the cell surface, and hence, increased dox and emodin accumulation and growth inhibition. If so, elevation of LRP1 expression may be a useful target for interventions to promote the efficacy of these and other anticancer drugs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  1. Kim TH, Shin YJ, Won AJ, Lee BM, Choi WS, Jung JH, Chung HY, Kim HS (2014) Resveratrol enhances chemosensitivity of doxorubicin in multidrug-resistant human breast cancer cells via increased cellular influx of doxorubicin. Biochim Biophys Acta 1840:615–625

    Article  CAS  PubMed  Google Scholar 

  2. Minotti G, Menna P, Salvatorelli E, Cairo G, Gianni L (2004) Anthracyclines: molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacol Rev 56:185–229

    Article  CAS  PubMed  Google Scholar 

  3. Esteva FJ, Valero V, Pusztai L, Boehnke-Michaud L, Buzdar AU, Hortobagyi GN (2001) Chemotherapy of metastatic breast cancer: what to expect in 2001 and beyond. Oncologist 6:133–146

    Article  CAS  PubMed  Google Scholar 

  4. Walker J, Martin C, Callaghan R (2004) Inhibition of P-glycoprotein function by XR9576 in a solid tumour model can restore anticancer drug efficacy. Eur J Cancer Oxf Engl 40:594–605

    Article  CAS  Google Scholar 

  5. Argov M, Kashi R, Peer D, Margalit R (2009) Treatment of resistant human colon cancer xenografts by a fluoxetine-doxorubicin combination enhances therapeutic responses comparable to an aggressive bevacizumab regimen. Cancer Lett 274:118–125

    Article  CAS  PubMed  Google Scholar 

  6. Scharenberg CW, Harkey MA, Torok-Storb B (2002) The ABCG2 transporter is an efficient Hoechst 33342 efflux pump and is preferentially expressed by immature human hematopoietic progenitors. Blood 99:507–512

    Article  CAS  PubMed  Google Scholar 

  7. Torky A-RW, Stehfest E, Viehweger K, Taege C, Foth H (2005) Immuno-histochemical detection of MRPs in human lung cells in culture. Toxicology 207:437–450

    Article  CAS  PubMed  Google Scholar 

  8. van der Deen M, de Vries EG, Timens W, Scheper RJ, Timmer-Bosscha H, Postma DS (2005) ATP-binding cassette (ABC) transporters in normal and pathological lung. Respir Res 6:59

    Article  PubMed  PubMed Central  Google Scholar 

  9. Borst P, Evers R, Kool M, Wijnholds J (1999) The multidrug resistance protein family. Biochim Biophys Acta 1461:347–357

    Article  CAS  PubMed  Google Scholar 

  10. Thews O, Gassner B, Kelleher DK, Gekle M (2008) Activity of drug efflux transporters in tumor cells under hypoxic conditions. Adv Exp Med Biol 614:157–164

    Article  CAS  PubMed  Google Scholar 

  11. Bellamy WT, Dalton WS, Kailey JM, Gleason MC, McCloskey TM, Dorr RT, Alberts DS (1988) Verapamil reversal of doxorubicin resistance in multidrug-resistant human myeloma cells and association with drug accumulation and DNA damage. Cancer Res 48:6365–6370

    CAS  PubMed  Google Scholar 

  12. Cole SP, Sparks KE, Fraser K, Loe DW, Grant CE, Wilson GM, Deeley RG (1994) Pharmacological characterization of multidrug resistant MRP-transfected human tumor cells. Cancer Res 54:5902–5910

    CAS  PubMed  Google Scholar 

  13. Folmer Y, Schneider M, Blum HE, Hafkemeyer P (2007) Reversal of drug resistance of hepatocellular carcinoma cells by adenoviral delivery of anti-ABCC2 antisense constructs. Cancer Gene Ther 14:875–884

    Article  CAS  PubMed  Google Scholar 

  14. Young LC, Campling BG, Cole SP, Deeley RG, Gerlach JH (2001) Multidrug resistance proteins MRP3, MRP1, and MRP2 in lung cancer: correlation of protein levels with drug response and messenger RNA levels. Clin Cancer Res Off J Am Assoc Cancer Res 7:1798–1804

    CAS  Google Scholar 

  15. Dandekar DS, Lokeshwar BL (2005) Emodin synergistically enhances docetaxel induced apoptosis in androgen-responsive prostate cancer cells. Cancer Res 65:723–723

    Google Scholar 

  16. Huang X, Wang J, Huang C, Chen Y, Shi G, Hu Q, Yi J (2008) Emodin enhances cytotoxicity of chemotherapeutic drugs in prostate cancer cells: the mechanisms involve ROS-mediated suppression of multidrug resistance and hypoxia inducible factor-1. Cancer Biol Ther 7:468–475

    Article  CAS  PubMed  Google Scholar 

  17. Wang W, Sun Y, Huang X, He M, Chen Y, Shi G, Li H, Yi J, Wang J (2010) Emodin enhances sensitivity of gallbladder cancer cells to platinum drugs via glutathion depletion and MRP1 downregulation. Biochem Pharmacol 79:1134–1140

    Article  CAS  PubMed  Google Scholar 

  18. Yu C-X, Zhang X-Q, Kang L-D, Zhang P-J, Chen W-W, Liu W-W, Liu Q-W, Zhang J-Y (2008) Emodin induces apoptosis in human prostate cancer cell LNCaP. Asian J Androl 10:625–634

    Article  CAS  PubMed  Google Scholar 

  19. Masaldan S, Iyer VV (2014) Exploration of effects of emodin in selected cancer cell lines: enhanced growth inhibition by ascorbic acid and regulation of LRP1 and AR under hypoxia-like conditions. J Appl Toxicol JAT 34:95–104

    Article  CAS  PubMed  Google Scholar 

  20. Kuo T-C, Yang J-S, Lin M-W, Hsu S-C, Lin J-J, Lin H-J, Hsia T-C, Liao C-L, Yang M-D, Fan M-J et al (2009) Emodin has cytotoxic and protective effects in rat C6 glioma cells: roles of Mdr1a and nuclear factor kappaB in cell survival. J Pharmacol Exp Ther 330:736–744

    Article  CAS  PubMed  Google Scholar 

  21. Li X, Wang H, Wang J, Chen Y, Yin X, Shi G, Li H, Hu Z, Liang X (2016) Emodin enhances cisplatin-induced cytotoxicity in human bladder cancer cells through ROS elevation and MRP1 downregulation. BMC Cancer 16:578

    Article  PubMed  PubMed Central  Google Scholar 

  22. Richard V, Nair MG, Kumar TRS, Pillai MR (2013) Side population cells as prototype of chemoresistant, tumor-initiating cells. BioMed Res Int 2013:e517237

    Article  Google Scholar 

  23. Huang Q, Shen H-M, Shui G, Wenk MR, Ong C-N (2006) Emodin inhibits tumor cell adhesion through disruption of the membrane lipid Raft-associated integrin signaling pathway. Cancer Res 66:5807–5815

    Article  CAS  PubMed  Google Scholar 

  24. Kawata K, Kubota S, Eguchi T, Aoyama E, Moritani NH, Kondo S, Nishida T, Takigawa M (2012) Role of LRP1 in transport of CCN2 protein in chondrocytes. J Cell Sci 125:2965–2972

    Article  CAS  PubMed  Google Scholar 

  25. Campana WM, Li X, Dragojlovic N, Janes J, Gaultier A, Gonias SL (2006) The low-density lipoprotein receptor-related protein is a pro-survival receptor in Schwann cells: possible implications in peripheral nerve injury. J Neurosci Off J Soc Neurosci 26:11197–11207

    Article  CAS  Google Scholar 

  26. Koong AC, Denko NC, Hudson KM, Schindler C, Swiersz L, Koch C, Evans S, Ibrahim H, Le QT, Terris DJ et al (2000) Candidate genes for the hypoxic tumor phenotype. Cancer Res 60:883–887

    CAS  PubMed  Google Scholar 

  27. Montel V, Gaultier A, Lester RD, Campana WM, Gonias SL (2007) The low-density lipoprotein receptor-related protein regulates cancer cell survival and metastasis development. Cancer Res 67:9817–9824

    Article  CAS  PubMed  Google Scholar 

  28. Newton CS, Loukinova E, Mikhailenko I, Ranganathan S, Gao Y, Haudenschild C, Strickland DK (2005) Platelet-derived growth factor receptor-β (PDGFR-β) activation promotes its association with the low density lipoprotein receptor-related protein (LRP) evidence for co-receptor function. J Biol Chem 280:27872–27878

    Article  CAS  PubMed  Google Scholar 

  29. Goldberg MA, Schneider TJ (1994) Similarities between the oxygen-sensing mechanisms regulating the expression of vascular endothelial growth factor and erythropoietin. J Biol Chem 269:4355–4359

    CAS  PubMed  Google Scholar 

  30. Shoemaker RH (2006) The NCI60 human tumour cell line anticancer drug screen. Nat Rev Cancer 6:813–823

    Article  CAS  PubMed  Google Scholar 

  31. Goren D, Horowitz AT, Tzemach D, Tarshish M, Zalipsky S, Gabizon A (2000) Nuclear delivery of doxorubicin via folate-targeted liposomes with bypass of multidrug-resistance efflux pump. Clin Cancer Res Off J Am Assoc Cancer Res 6:1949–1957

    CAS  Google Scholar 

  32. Al-Nuri MA, Za’tar NA, Abu-Eid MA, Hannoun MA, Al-Jondi WJ, Hussein AI, Ali-Shtayeh MS (1996) Emodin, a naturally occuring anthraquinone: its isolation and spectrophotometric determination in rumex cyprius plant. Spectrosc Lett 29:1539–1543

    Article  CAS  Google Scholar 

  33. Barenolz Y, Haran G (1993). Method of amphiphatic drug loading in liposomes by pH gradient (Google Patents). http://www.google.co.in/patents/US5192549

  34. Hartree EF (1972) Determination of protein: a modification of the Lowry method that gives a linear photometric response. Anal Biochem 48:422–427

    Article  CAS  PubMed  Google Scholar 

  35. Ying Y, Qingwu L, Mingming X, Zhenju S, Chaoyang T, Zhengang T (2017) Emodin: one main ingredient of Shufeng Jiedu capsule reverses chemoresistance of lung cancer cells through inhibition of EMT. Cell Physiol Biochem Int J Exp Cell Physiol Biochem Pharmacol 42:1063–1072

    Article  Google Scholar 

  36. Li X, Dong Y, Wang W, Wang H, Chen Y, Shi G, Yi J, Wang J (2013) Emodin as an effective agent in targeting cancer stem-like side population cells of gallbladder carcinoma. Stem Cells Dev 22:554–566

    Article  CAS  PubMed  Google Scholar 

  37. Wei W-T, Lin S-Z, Liu D-L, Wang Z-H (2013) The distinct mechanisms of the antitumor activity of emodin in different types of cancer (Review). Oncol Rep 30:2555–2562

    Article  CAS  PubMed  Google Scholar 

  38. Drug combinations to overcome treatment resistance. National Cancer Institute. https://www.cancer.gov/about-cancer/treatment/research/drug-combo-resistance. Accessed 15 Feb 2018

  39. Mokhtari RB, Homayouni TS, Baluch N, Morgatskaya E, Kumar S, Das B, Yeger H (2017) Combination therapy in combating cancer. Oncotarget 8:38022–38043

    Article  PubMed Central  Google Scholar 

  40. Acton QA (2012) ATP-binding cassette transporters: advances in research and application. 2011 Edition: ScholarlyBrief (ScholarlyEditions). https://books.google.co.in/books?id=8ExP72uUt-kC

Download references

Acknowledgements

The authors thank Dr. K.G. Thirumurugaan (Translational Research Platform for Veterinary Biologicals, Tamil Nadu Veterinary and Animal Sciences University) for his support in confocal microscopy. We are also grateful to Ms. Kavitha Murugan for her help with the revision of figures. This study and YP (as Junior Research Fellow) were supported financially through SB/YS/LS-138/2013 from Science Engineering and Research Board (SERB), Government of India.

Author information

Author notes

  1. Vidhya V. Iyer

    Present address: Department of Chemical Engineering, Indian Institute of Technology Bombay Powai, Mumbai, Maharashtra, 400076, India

  2. P. Yoga Priya and Jeipreeti Kangeyavelu have contributed equally to the work.

Authors and Affiliations

  1. School of Biosciences and Technology, Vellore Institute of Technology, Vellore, Tamil Nadu, 632 014, India

    Vidhya V. Iyer, P. Yoga Priya & Jeipreeti Kangeyavelu

  2. Madurai, India

    P. Yoga Priya

  3. Chennai, India

    Jeipreeti Kangeyavelu

Authors
  1. Vidhya V. Iyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Yoga Priya
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jeipreeti Kangeyavelu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conceived and designed the experiments: VI; performed the experiments: JK (GI, qPCR), YP (accumulation, qPCR); analyzed the data: VI, JK, YP; wrote the paper: VI.

Corresponding author

Correspondence to Vidhya V. Iyer.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest in this study.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Iyer, V.V., Priya, P.Y. & Kangeyavelu, J. Effects of increased accumulation of doxorubicin due to emodin on efflux transporter and LRP1 expression in lung adenocarcinoma and colorectal carcinoma cells. Mol Cell Biochem 449, 91–104 (2018). https://doi.org/10.1007/s11010-018-3346-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-018-3346-4

Keywords

Search

Navigation