Skip to main content

Advertisement

SpringerLink
Log in

Schisandrin A reverses doxorubicin-resistant human breast cancer cell line by the inhibition of P65 and Stat3 phosphorylation

  • Original Article
  • Published:
Breast Cancer Aims and scope Submit manuscript

Abstract

Background

Multidrug resistance (MDR) in breast cancer therapy occurs frequently. Thus, anti-MDR agents from natural products or synthetic compounds were tested extensively. We have also explored the reverse effect and mechanism of Schisandrin A (Sch A), a natural product, on MCF-7 breast cancer doxorubicin (DOX)-resistant subline MCF-7/DOX.

Methods

MTT assay was performed to measure the viability of MCF-7 cells to assess the reverse effect of Sch A. Western blot analysis was used to study the protein levels. Laser scanning confocal microscopy was performed to detect the intercellular DOX and Rhodamine 123 accumulation. The qRT-PCR was used to analysis the target gene expression. Dual-luciferase reporter assay was performed to test the transcriptional activity of P-glycoprotein (P-gp).

Results

Sch A, at the concentration of 20 µM, showed selective reverse effect (better than the positive control, verapamil at 5 µM) on MCF-7/DOX cell line but not on BEL-7402/DOX, Hep G2/DOX, and K-562/DOX cells. In addition, Sch A enhanced DOX-induced cleavage of Caspase-9 and PARP levels by increasing intracellular DOX accumulation and inhibiting P-gp function. Furthermore, Sch A selectively suppressed P-gp at gene and protein levels in MCF-7/DOX cells which express high level of MDR1 but not MRP1, MRP3, or BCRP. Besides, Sch A showed inhibitory effect on P-gp transcriptional activity. Sch A significantly reduced p-IκB-α (Ser32) and p-Stat3 (Tyr705) levels which mediate P-gp expression. In addition, Stat3 knockdown enhanced the reverse effect of siP65. The combined effect of siStat3 and siP65 was better than Sch A single treatment in MCF-7/DOX cells.

Conclusion

Sch A specifically reverses P-gp-mediated DOX resistance in MCF-7/DOX cells by blocking P-gp, NF-κB, and Stat3 signaling. Inhibition of P65 and Stat3 shows potent anti-MDR effect on MCF-7/DOX cells.

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

Similar content being viewed by others

References

  1. Huang SD, Holzel M, Knijnenburg T, Schlicker A, Roepman P, McDermott U, et al. MED12 controls the response to multiple cancer drugs through regulation of TGF-beta receptor signaling. Cell. 2012;151:937–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Lee C, Park S, Kim JH, Lim SM, Park HS, Kim SI, et al. Expression of T-lymphocyte markers in human epidermal growth factor receptor 2-positive breast cancer. J Breast Cancer. 2016;19:385–93.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Xia YZ, Ni K, Guo C, Zhang C, Geng YD, Wang ZD, et al. Alopecurone B reverses doxorubicin-resistant human osteosarcoma cell line by inhibiting P-glycoprotein and NF-kappa B signaling. Phytomedicine. 2015;22:344–51.

    Article  CAS  PubMed  Google Scholar 

  4. Zhao XQ, Xie JD, Chen XG, Sim HM, Zhang X, Liang YJ, et al. Neratinib reverses ATP-binding cassette B1-mediated chemotherapeutic drug resistance in vitro, in vivo, and ex vivo. Mol Pharmacol. 2012;82:47–58.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Gillet JP, Gottesman MM. Overcoming multidrug resistance in cancer: 35 years after the discovery of ABCB1. Drug Resist Updat. 2012;15:2–4.

    Article  PubMed  Google Scholar 

  6. Kim SJ, Min HY, Lee EJ, Kim YS, Bae K, Kang SS, et al. Growth inhibition and cell cycle arrest in the G0/G1 by schizandrin, a dibenzocyclooctadiene lignan isolated from Schisandra chinensis, on T47D human breast cancer cells. Phytother Res. 2010;24:193–7.

    CAS  PubMed  Google Scholar 

  7. Xia YZ, Yang L, Wang ZD, Guo C, Zhang C, Geng YD, et al. Schisandrin A enhances the cytotoxicity of doxorubicin by the inhibition of nuclear factor-kappa B signaling in a doxorubicin-resistant human osteosarcoma cell line. RSC Adv. 2015;5:13972–84.

    Article  CAS  Google Scholar 

  8. Wang LJ, Meng Q, Wang CY, Liu Q, Peng JY, Huo XK, et al. Dioscin restores the activity of the anticancer agent adriamycin in multidrug-resistant human leukemia K562/adriamycin cells by down-regulating MDR1 via a mechanism involving NF-kappa B signaling inhibition. J Nat Prod. 2013;76:909–14.

    Article  CAS  PubMed  Google Scholar 

  9. Lu Y, Li F, Xu T, Sun J. Tetrandrine prevents multidrug resistance in the osteosarcoma cell line, U-2OS, by preventing Pgp overexpression through the inhibition of NF-kappaB signaling. Int J Mol Med. 2017;39:993–1000.

    Article  PubMed  Google Scholar 

  10. Assef Y, Rubio F, Colo G, del Monaco S, Costas MA, Kotsias BA. Imatinib resistance in multidrug-resistant K562 human leukemic cells. Leuk Res. 2009;33:710–6.

    Article  CAS  PubMed  Google Scholar 

  11. Watanabe A, Yamamoto K, Ioroi T, Hirata S, Harada K, Miyake H, et al. Association of single nucleotide polymorphisms in STAT3, ABCB1, and ABCG2 with stomatitis in patients with metastatic renal cell carcinoma treated with sunitinib: a retrospective analysis in Japanese patients. Biol Pharm Bull. 2017;40:458–64.

    Article  CAS  PubMed  Google Scholar 

  12. Zhang F, Wang Z, Fan Y, Xu Q, Ji W, Tian R, et al. Elevated STAT3 signaling-mediated upregulation of MMP-2/9 confers enhanced invasion ability in multidrug-resistant breast cancer cells. Int J Mol Sci. 2015;16:24772–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Johnston PA, Grandis JR. STAT3 signaling: anticancer strategies and challenges. Mol Interv. 2011;11:18–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Chiu LY, Ko JL, Lee YJ, Yang TY, Tee YT, Sheu GT. L-type calcium channel blockers reverse docetaxel and vincristine-induced multidrug resistance independent of ABCB1 expression in human lung cancer cell lines. Toxicol Lett. 2010;192:408–18.

    Article  CAS  PubMed  Google Scholar 

  15. Shi Z, Liang YJ, Chen ZS, Wang XW, Wang XH, Ding Y, et al. Reversal of MDR1/P-glycoprotein-mediated multidrug resistance by vector-based RNA interference in vitro and in vivo. Cancer Biol Ther. 2006;5:39–47.

    Article  CAS  PubMed  Google Scholar 

  16. Xia YZ, Yang L, Xue GM, Zhang C, Guo C, Yang YW, et al. Combining GRP78 suppression and MK2206-induced Akt inhibition decreases doxorubicin-induced P-glycoprotein expression and mitigates chemoresistance in human osteosarcoma. Oncotarget. 2016;7:56371–82.

    PubMed  PubMed Central  Google Scholar 

  17. Zhu FF, Dai CY, Fu YF, Loo JFC, Xia DJ, Gao SP, et al. Physalin A exerts anti-tumor activity in non-small cell lung cancer cell lines by suppressing JAK/STAT3 signaling. Oncotarget. 2016;7:9463–77.

    Google Scholar 

  18. Berger W, Setinek U, Hollaus P, Zidek T, Steiner E, Elbling L, et al. Multidrug resistance markers P-glycoprotein, multidrug resistance protein 1, and lung resistance protein in non-small cell lung cancer: prognostic implications. J Cancer Res Clin Oncol. 2005;131:355–63.

    Article  CAS  PubMed  Google Scholar 

  19. Bruhn O, Cascorbi I. Polymorphisms of the drug transporters ABCB1, ABCG2, ABCC2 and ABCC3 and their impact on drug bioavailability and clinical relevance. Expert Opin Drug Metab Toxicol. 2014;10:1337–54.

    Article  CAS  PubMed  Google Scholar 

  20. Robey RW, Ierano C, Zhan Z, Bates SE. The challenge of exploiting ABCG2 in the clinic. Curr Pharm Biotechnol. 2011;12:595–608.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Guo Y, Ding Y, Zhang T, An H. Sinapine reverses multi-drug resistance in MCF-7/dox cancer cells by downregulating FGFR4/FRS2alpha-ERK1/2 pathway-mediated NF-kappaB activation. Phytomedicine. 2016;23:267–73.

    Article  CAS  PubMed  Google Scholar 

  22. Liu J, Zhou F, Chen Q, Kang A, Lu M, Liu W, et al. Chronic inflammation up-regulates P-gp in peripheral mononuclear blood cells via the STAT3/Nf-kappab pathway in 2,4,6-trinitrobenzene sulfonic acid-induced colitis mice. Sci Rep. 2015;5:13558.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Krishna R, Mayer LD. Multidrug resistance (MDR) in cancer. Mechanisms, reversal using modulators of MDR and the role of MDR modulators in influencing the pharmacokinetics of anticancer drugs. Eur J Pharm Sci. 2000;11:265–83.

    Article  CAS  PubMed  Google Scholar 

  24. Gill J, Ahluwalia MK, Geller D, Gorlick R. New targets and approaches in osteosarcoma. Pharmacol Ther. 2013;137:89–99.

    Article  CAS  PubMed  Google Scholar 

  25. Campbell KJRS, Perkins ND. Active repression of antiapoptotic gene expression by RelA(p65) NF-kappa B. Mol Cell. 2004;13:853–65.

    Article  CAS  PubMed  Google Scholar 

  26. Narayanan PD, Nandabalan SK, Baddireddi LS. Role of STAT3 phosphorylation in ethanol-mediated proliferation of breast cancer cells. J Breast Cancer. 2016;19:122–32.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pharmacy, Linyi People’s Hospital, Linyi, Shandong, China

    Zong-Lin Zhang

  2. Department of Pharmacy, First People’s Hospital of Tancheng County, Tancheng, Shandong, China

    Qing-Cheng Jiang

  3. Department of Gynecology and Obstetrics, Linyi People’s Hospital, 27# Jie fang lu dong duan, Linyi, Shandong, China

    Su-Rong Wang

Authors
  1. Zong-Lin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qing-Cheng Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Su-Rong Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Su-Rong Wang.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, ZL., Jiang, QC. & Wang, SR. Schisandrin A reverses doxorubicin-resistant human breast cancer cell line by the inhibition of P65 and Stat3 phosphorylation. Breast Cancer 25, 233–242 (2018). https://doi.org/10.1007/s12282-017-0821-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12282-017-0821-9

Keywords

Search

Navigation