Skip to main content

Advertisement

SpringerLink
Log in

Differential effects of c-myc and ABCB1 silencing on reversing drug resistance in HepG2/Dox cells

  • Original Article
  • Published:
Tumor Biology

Abstract

Multidrug resistance (MDR) in various kinds of cancers represents a true obstacle which hinders the successes of most of current available chemotherapies. ATP-binding cassette (ABC) trasporter proteins have been shown to contribute to the majority of MDR in various types of malignancies. c-myc has recently been reported to participate, at least partly, in MDR to some types of cancers. This study aimed to test whether c-myc could play a role, solely or with coordination with other ABCs, in the resistance of HepG2 cells to doxorubicin (Dox). MDR has been induced in wild-type HepG2 and has been verified both on gene and protein levels. Various assays including efflux assays as well as siRNA targeting ABCB1 and c-myc have been employed to explore the role of both candidate molecules in MDR in HepG2. Results obtained, with regard to ABCB1 silencing on HepG2/Dox cells, have shown that ABCB1-deficient cells exhibited a significant reduction in ABCC1 expression as compared to ABCB1-sufficient cells. However, these cells did not show a significant reduction in other tested ABCs (ABCC5 and ABCC10) while c-myc silencing had no significant effect on any of the studied ABCs. Moreover, silencing of ABCB1 on HepG2 significantly increased fluorescent calcein retention in HepG2 cells as compared to the control cells while downregulation of c-myc did not have any effect on fluorescent calcein retention. Altogether, this work clearly demonstrates that c-myc has no role in MDR of HepG2 to Dox which has been shown to be ABCB1-mediated in a mechanism which might involve ABCC1.

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.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Gottesman MM, Fojo T, Bates SE. Multidrug resistance in cancer: role of ATP dependent transporters. Nat Rev Cancer. 2002;2:48–58.

    Article  CAS  PubMed  Google Scholar 

  2. Gong J, Jaiswal R, Mathys JM, Combes V, Grau G, Bebawy M. Microparticles and their emerging role in cancer multidrug resistance. Cancer Treat Rev. 2012;38:226–34.

    Article  CAS  PubMed  Google Scholar 

  3. Wang Z, Xu Y, Meng X, Watari F, Liu H. Suppression of c-Myc is involved in multi-walled carbon nanotubes’ down-regulation of ATP-binding cassette transporters in human colon adenocarcinoma cells. Toxicol Appl Pharmacol. 2015;282(1):42–51.

    Article  CAS  PubMed  Google Scholar 

  4. Kim D, Kim M, Kim H, Leea J, Baea J, Kim D, et al. Suppression of multidrug resistance by treatment with TRAIL in human ovarian and breast cancer cells with high level of c-Myc. Biochim Biophys Acta. 1812;2011:796–805.

    Google Scholar 

  5. Sumi TN, Tsuneyoshi N, Nakatsuji N, Suemori H. Apoptosis and differentiation of human embryonic stem cells induced by sustained activation of c-Myc. Oncogene. 2007;26:5564–76.

    Article  CAS  PubMed  Google Scholar 

  6. Hoffman B, Liebermann DA. Apoptotic signaling by c-MYC. Oncogene. 2008;27:6462–72.

    Article  CAS  PubMed  Google Scholar 

  7. Porro A, Iraci N, Soverini S, Diolaiti D, Gherardil S, Terragna C, et al. c-MYC oncoprotein dictates transcriptional profiles of ATP-binding cassette transporter genes in chronic myelogenous leukemia CD34 hematopoietic progenitor cells. Mol Cancer Res. 2011;9(8):1054–66.

    Article  CAS  PubMed  Google Scholar 

  8. Kugimiya N, Nishimoto A, Hosoyama T, Ueno K, Enoki T, Li TS, et al. The c-MYC-ABCB5 axis plays a pivotal role in 5-fluorouracil resistance in human colon cancer cells. J Cell Mol Med. 2015. doi:10.1111/jcmm.12531.

    PubMed  PubMed Central  Google Scholar 

  9. Tiwaria AK, Sodania K, Daia C, Abuznaitb AH, Singha S, Xiaoa Z, et al. Nilotinib potentiates anticancer drug sensitivity in murine ABCB1-, ABCG2-, and ABCC10-multidrug resistance xenograft models Amit K. Chena Z. Cancer Lett. 2013;328(2):307–17.

    Article  Google Scholar 

  10. Zhang X, Yashiro M, Qiu H, Nishii T, Matsuzaki T, Hirakawa K. The role of c-myc in regulating mdr1 gene expression in tumor cell line KB. Anticancer Res. 2010;30:915–21.

    PubMed  Google Scholar 

  11. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-Dye binding. Anal Biochem. 1976;72:248–54.

    Article  CAS  PubMed  Google Scholar 

  12. Tiberghien F, Loor F. Ranking of P-glycoprotein substrates and inhibitors by a calcein-AM fluorometry screening assay. Anticancer Drug. 1996;7(5):568–78.

    Article  CAS  Google Scholar 

  13. Tiwari AK, Sodani K, Chen ZS. Current advances in modulation of ABC transporter-mediated multidrug resistance in cancer. Int J Toxicol Pharmcol Res. 2009;1:1–6.

    Google Scholar 

  14. Tsuruo T, Naito M, Tomida A, Fujita N, Mashima T, Sakamoto H, et al. Molecular targeting therapy of cancer: drug resistance, apoptosis and survival signal. Cancer Sci. 2003;94:15–21.

    Article  CAS  PubMed  Google Scholar 

  15. Wang X, Campos CR, Peart JC, Smith LK, Boni JL, Cannon RE, et al. Nrf2 upregulates ATP binding cassette transporter expression and activity at the blood–brain and blood–spinal cord barriers. J Neurosci. 2014;34(25):8585–93.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Lee CH. Reversing agents for ATP-binding cassette (ABC) transporters: application in modulating multidrug resistance (MDR). Curr Med Chem Anticancer Agents. 2004;4:43–52.

    Article  CAS  PubMed  Google Scholar 

  17. Leonessa F, Clarke R. ATP binding cassette transporters and drug resistance in breast cancer. Endocr Relat Cancer. 2003;10:43–73.

    Article  CAS  PubMed  Google Scholar 

  18. Albihn A, Johnsen JI, Henriksson MA. MYC in oncogenesis and as a target for cancer therapies. Adv Cancer Res. 2010;107:163–224.

    Article  CAS  PubMed  Google Scholar 

  19. Nieminen AI, Partanen JI, Klefstrom J. c-Myc blazing a trail of death: coupling of the mitochondrial and death receptor apoptosis pathways by c-Myc. Cell Cycle. 2007;6:2464–72.

    Article  CAS  PubMed  Google Scholar 

  20. He Y, Zhang J, et al. The role of c-myc in regulating mdr1 gene expression in tumor cell line KB. Chin Med J. 2000;113(9):848–51.

    CAS  PubMed  Google Scholar 

  21. Yang X, Cai H, et al. Inhibition of c-Myc by let-7b mimic reverses mutidrug resistance in gastric cancer cells. Oncol Rep. 2015;33(4):1723–30.

    CAS  PubMed  Google Scholar 

  22. Xia C, Tian C, et al. c-Myc plays part in drug resistance mediated by bone marrow stromal cells in acute myeloid leukemia. Leuk Res. 2014;39(1):92–9.

    Article  PubMed  Google Scholar 

  23. Borel F, Logtenstein R, Koornneef A, Maczuga P, Ritsema T, Petry H, et al. In vivo knock-down of multidrug resistance transporters ABCC1 and ABCC2 by AAV-delivered shRNAs and by artificial miRNAs. J RNAi Gene Silenc. 2011;7:434–42.

    CAS  Google Scholar 

  24. Qiu J, Zhang Y, Li Y, Zhao Y, Zhang W, Jiang Q, et al. Trametinib modulates cancer multidrug resistance by targeting ABCB1 transporter. Oncotargets. 2015;6(17):15495–509.

    Google Scholar 

  25. Cole SP, Bhardwaj G, Gerlach JH, Mackie JE, Grant CE, Almquist KC, et al. Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line. Science. 1992;258:1650–4.

    Article  CAS  PubMed  Google Scholar 

  26. Kubota T, Furukawa H, Tanino A, Suto Y, Otan M, Watanabe T, et al. Resistant mechanisms of anthracyclines: pirarubicin might partly break through the P-glycoprotein-mediated drugresistance of human breast cancer tissues. Breast Cancer. 2001;8:333–8.

    Article  CAS  PubMed  Google Scholar 

  27. Nooter K, Westerman AM, Flens MJ, Zaman GJ, Scheper RJ, van Wingerden KE, et al. Expression of the multidrug resistance- associated protein (MRP) gene in human cancers. Clin Cancer Res. 1995;1:1301–10.

    CAS  PubMed  Google Scholar 

  28. Flens MJ, Zaman GJ, van der Valk P, Izquierdo MA, Schroeijers AB, Scheffer LG, et al. Tissue distribution of the multidrug resistance protein. Am J Pathol. 1996;148:1237–47.

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Chen ZS, Tiwari AK. Multidrug resistance proteins (MRPs/ABCCs) in cancer chemotherapy and genetic diseases. FEBS J. 2011;278:3226–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Deeley RG, Westlake C, Cole SP. Transmembrane transport of endo- and xenobiotics by mammalian ATP-binding cassette multidrug resistance proteins. Physiol Rev. 2006;86:849–899.29.

    Article  CAS  PubMed  Google Scholar 

  31. Sau A, PellizzariTregno F, Valentino F, Federici G, Caccuri AM. Glutathione transferases and development of new principles to overcome drug resistance. Arch Biochem Biophys. 2010;500:116–22.

    Article  CAS  PubMed  Google Scholar 

  32. Mao Q, Deeley RG, Cole SP. Functional reconstitution of substrate transport by purified multidrug resistance protein MRP1 (ABCC1) in phospholipid vesicles. J Biol Chem. 2000;275:34166–72.

    Article  CAS  PubMed  Google Scholar 

  33. Wang L, Deng Q, Wang J, Bai X, Xiao X, Hai-Rong LV, et al. Effect of CIK on multidrug-resistance reversal and increasing the sensitivity of ADR in K562/ADR cells. Oncol Lett. 2014;8(4):1778–82.

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the Science and Technology Development Fund (STDF), Egypt, Basic and Applied Research Grant (Project ID 4361). We thank Ghada H. El-sayed, Heba K. Nabih, Eman M. Abdel Motaleb, and Alaa Shahin for technical help.

Author information

Authors and Affiliations

  1. Hormones Department, Meical Division, National Research Centre, 33 El Bohouth st. (Former El Tahrir st.) Dokki, P.O. 12622, Giza, Egypt

    Shaymaa M. M. Yahya

  2. Pharmaceutical Research Group, Center of Excellence for Advanced Sciences and Phytochemistry Department National Research Centre, 33 El Bohouth st. (Former El Tahrir st) Dokki, P.O. 12622, Giza, Egypt

    Ahmed R. Hamed & Maha M. Soltan

  3. Department of Microbiology and Immunology, Faculty of Pharmacy, Helwan University, Ein Helwan, P.O.X: 11795, Cairo, Egypt

    Mohamed Emara

  4. Pharmaceutical and Drug Industries Research Division, Therapeutic Chemistry Department, National Research Center, 33 El Bohouth st. (Former El Tahrir st) Dokki, P.O. 12622, Giza, Egypt

    Gamal Eldein F. Abd-Ellatef

  5. Microbial Biotechnology Department, National Research Center, 33 El Bohouth st. (Former El Tahrir st) Dokki, P.O. 12622, Giza, Egypt

    Salma M. Abdelnasser

Authors
  1. Shaymaa M. M. Yahya
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ahmed R. Hamed
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mohamed Emara
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maha M. Soltan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gamal Eldein F. Abd-Ellatef
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Salma M. Abdelnasser
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shaymaa M. M. Yahya.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yahya, S.M.M., Hamed, A.R., Emara, M. et al. Differential effects of c-myc and ABCB1 silencing on reversing drug resistance in HepG2/Dox cells. Tumor Biol. 37, 5925–5932 (2016). https://doi.org/10.1007/s13277-015-4426-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13277-015-4426-7

Keywords

Search

Navigation