Molecular and Cellular Biochemistry

, Volume 394, Issue 1–2, pp 209–215 | Cite as

Berberine diminishes side population and down-regulates stem cell-associated genes in the pancreatic cancer cell lines PANC-1 and MIA PaCa-2

  • S. H. Park
  • J. H. Sung
  • N. ChungEmail author


Cancer stem cells play an important role in metastasis and the relapse of drug resistant cancers. Side-population (SP) cells are capable of effluxing Hoechst 33342 dye and are referred to as cancer stem cells. We investigated the effect of berberine on pancreatic cancer stem cells of PANC-1 and MIA PaCa-2. For both cell lines, the proportions of SP cells in the presence of berberine were investigated and compared to the proportions in the presence of gemcitabine, a standard pancreatic anti-cancer drug. The proportions of SP cells in the PANC-1 and MIA PaCa-2 cell lines were about 9 and <0.1 %, respectively. After berberine and gemcitabine treatments, the SP cell proportion of PANC-1 decreased to 5.7 ± 2.0 and 6.8 ± 0.8 %, respectively, which compares to the control proportion of (9.7 ± 1.7). After berberine and gemcitabine treatment of PANC-1, of the four stem cell-associated genes (SOX2, POU5F1, NANOG, and NOTCH1), all but NOTCH1 were down-regulated. Unfortunately, the effect of berberine and gemcitabine treatments on MIA PaCa-2 SP cells could not be clearly observed because SP cells represented only a very small proportion of MIA PaCa-2 cells. However, SOX2, POU5F1, and NANOG genes were shown to be effectively down-regulated in the MIA PaCa-2 cell line as a whole. Taken together, these results indicate that berberine is as effective at targeting pancreatic cancer cell lines as gemcitabine. Therefore, we believe that POU5F1, SOX2, and NANOG can serve as potential markers, and berberine may be an effective anti-cancer agent when targeting human pancreatic cancer cells and/or their cancer stem cells.


Pancreatic cancer Cancer stem cells Side population Berberine POU5F1 NANOG 



This study was supported by a Grant (B110053) from the Korea Health Technology R&D Project, Ministry of Health and Welfare, and a grant from Korea University.

Conflict of interest

The authors declare that no conflicts of interest exist.


  1. 1.
    Burkert J, Wright NA, Alison MR (2006) Stem cells and cancer: an intimate relationship. J Pathol 209:287–297. doi: 10.1002/path.2016 PubMedCrossRefGoogle Scholar
  2. 2.
    Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE (2007) Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci USA 104:973–978. doi: 10.1073/pnas.0610117104 PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Dalerba P, Cho RW, Clarke MF (2007) Cancer stem cells: models and concepts. Annu Rev Med 58:267–284. doi: 10.1146/ PubMedCrossRefGoogle Scholar
  4. 4.
    Zhou S, Schuetz JD, Bunting KD, Colapietro AM, Sampath J, Morris JJ, Lagutina I, Grosveld GC, Osawa M, Nakauchi H, Sorrentino BP (2001) The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. Nat Med 7:1028–1034. doi: 10.1038/nm0901-1028 PubMedCrossRefGoogle Scholar
  5. 5.
    Wu C, Alman BA (2008) Side population cells in human cancers. Cancer Lett 268:1–9. doi: 10.1016/j.canlet.2008.03.048 PubMedCrossRefGoogle Scholar
  6. 6.
    Challen GA, Little MH (2006) A side order of stem cells: the SP phenotype. Stem Cells 24:3–12. doi: 10.1634/stemcells.2005-0116 PubMedCrossRefGoogle Scholar
  7. 7.
    Donnenberg VS, Landreneau RJ, Donnenberg AD (2007) Tumorigenic stem and progenitor cells: implications for the therapeutic index of anti-cancer agents. J Control Release 122:385–391. doi: 10.1016/j.jconrel.2007.05.005 PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Kondo T, Setoguchi T, Taga T (2004) Persistence of a small subpopulation of cancer stem-like cells in the C6 glioma cell line. Proc Natl Acad Sci USA 101:781–786. doi: 10.1073/pnas.0307618100 PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Wang J, Guo LP, Chen LZ, Zeng YX, Lu SH (2007) Identification of cancer stem cell-like side population cells in human nasopharyngeal carcinoma cell line. Cancer Res 67:3716–3724. doi: 10.1158/0008-5472.CAN-06-4343 PubMedCrossRefGoogle Scholar
  10. 10.
    Li C, Heidt DG, Dalerba P, Burant CF, Zhang L, Adsay V, Wicha M, Clarke MF, Simeone DM (2007) Identification of pancreatic cancer stem cells. Cancer Res 67:1030–1037. doi: 10.1158/0008-5472.CAN-06-2030 PubMedCrossRefGoogle Scholar
  11. 11.
    Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ (2009) Cancer statistics, 2009. CA Cancer J Clin 59:225–249. doi: 10.3322/caac.20006 PubMedCrossRefGoogle Scholar
  12. 12.
    Wang SJ, Gao Y, Chen H, Kong R, Jiang HC, Pan SH, Xue DB, Bai XW, Sun B (2010) Dihydroartemisinin inactivates NF-kappaB and potentiates the anti-tumor effect of gemcitabine on pancreatic cancer both in vitro and in vivo. Cancer Lett 293:99–108. doi: 10.1016/j.canlet.2010.01.001 PubMedCrossRefGoogle Scholar
  13. 13.
    O’’eilly EM, Abou-Alfa GK (2007) Cytotoxic therapy for advanced pancreatic adenocarcinoma. Semin Oncol 34:347–353. doi: 10.1053/j.seminoncol.2007.05.009 CrossRefGoogle Scholar
  14. 14.
    Bezakova L, Misik V, Malekova L, Svajdlenka E, Kostalova D (1996) Lipoxygenase inhibition and antioxidant properties of bisbenzylisoqunoline alkaloids isolated from Mahonia aquifolium. Pharmazie 51:758–761PubMedGoogle Scholar
  15. 15.
    Misik V, Bezakova L, Malekova L, Kostalova D (1995) Lipoxygenase inhibition and antioxidant properties of protoberberine and aporphine alkaloids isolated from Mahonia aquifolium. Planta Med 61:372–373PubMedCrossRefGoogle Scholar
  16. 16.
    Chen J, Zhao H, Wang X, Lee FS, Yang H, Zheng L (2008) Analysis of major alkaloids in Rhizoma coptidis by capillary electrophoresis–electrospray–time of flight mass spectrometry with different background electrolytes. Electrophoresis 29:2135–2147. doi: 10.1002/elps.200700797 PubMedCrossRefGoogle Scholar
  17. 17.
    Zhu XZ, Li XY, Liu J (2004) Recent pharmacological studies on natural products in China. Eur J Pharmacol 500:221–230. doi: 10.1016/j.ejphar.2004.07.027 PubMedCrossRefGoogle Scholar
  18. 18.
    Mantena SK, Sharma SD, Katiyar SK (2006) Berberine, a natural product, induces G1-phase cell cycle arrest and caspase-3-dependent apoptosis in human prostate carcinoma cells. Mol Cancer Ther 5:296–308. doi: 10.1158/1535-7163.MCT-05-0448 PubMedCrossRefGoogle Scholar
  19. 19.
    Kim JB, Ko E, Han W, Shin I, Park SY, Noh DY (2008) Berberine diminishes the side population and ABCG2 transporter expression in MCF-7 breast cancer cells. Planta Med 74:1693–1700. doi: 10.1055/s-0028-1088313 PubMedCrossRefGoogle Scholar
  20. 20.
    Siegel R, Naishadham D, Jemal A (2012) Cancer statistics for Hispanics/Latinos, 2012. CA Cancer J Clin 62. doi: 10.3322/caac.21153
  21. 21.
    Zhou J, Wulfkuhle J, Zhang H, Gu P, Yang Y, Deng J, Margolick JB, Liotta LA, Petricoin E 3rd, Zhang Y (2007) Activation of the PTEN/mTOR/STAT3 pathway in breast cancer stem-like cells is required for viability and maintenance. Proc Natl Acad Sci USA 104:16158–16163. doi: 10.1073/pnas.0702596104 PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Patrawala L, Calhoun T, Schneider-Broussard R, Zhou J, Claypool K, Tang DG (2005) Side population is enriched in tumorigenic, stem-like cancer cells, whereas ABCG2+ and ABCG2− cancer cells are similarly tumorigenic. Cancer Res 65:6207–6219. doi: 10.1158/0008-5472.CAN-05-0592 PubMedCrossRefGoogle Scholar
  23. 23.
    Wu C, Wei Q, Utomo V, Nadesan P, Whetstone H, Kandel R, Wunder JS, Alman BA (2007) Side population cells isolated from mesenchymal neoplasms have tumor initiating potential. Cancer Res 67:8216–8222. doi: 10.1158/0008-5472.CAN-07-0999 PubMedCrossRefGoogle Scholar
  24. 24.
    Tang Y, Kitisin K, Jogunoori W, Li C, Deng CX, Mueller SC, Ressom HW, Rashid A, He AR, Mendelson JS, Jessup JM, Shetty K, Zasloff M, Mishra B, Reddy EP, Johnson L, Mishra L (2008) Progenitor/stem cells give rise to liver cancer due to aberrant TGF-beta and IL-6 signaling. Proc Natl Acad Sci USA 105:2445–2450. doi: 10.1073/pnas.0705395105 PubMedCentralPubMedCrossRefGoogle Scholar
  25. 25.
    Miao (2010) Side population in the pancreatic cancer cell lines SW1990 and CFPAC-1 is enriched with cancer stem-like cells. Oncol Rep 23. doi: 10.3892/or_00000774
  26. 26.
    Sung JH, Kim JB, Park SH, Park SY, Lee JK, Lee H-S, Chung N (2012) Berberine decreases cell growth but increases the side population fraction of H460 lung cancer cells. J Korean Soc Appl Biol Chem 55:491–495. doi: 10.1007/s13765-012-2119-0
  27. 27.
    Rizzino A (2009) Sox2 and Oct-3/4: a versatile pair of master regulators that orchestrate the self-renewal and pluripotency of embryonic stem cells. Wiley Interdiscip Rev Syst Biol Med 1:228–236. doi: 10.1002/wsbm.12 PubMedCentralPubMedCrossRefGoogle Scholar
  28. 28.
    Takeda J, Seino S, Bell GI (1992) Human Oct3 gene family: cDNA sequences, alternative splicing, gene organization, chromosomal location, and expression at low levels in adult tissues. Nucleic Acids Res 20:4613–4620PubMedCentralPubMedCrossRefGoogle Scholar
  29. 29.
    Chambers I, Colby D, Robertson M, Nichols J, Lee S, Tweedie S, Smith A (2003) Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113:643–655PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Biosystems and Biotechnology, College of Life Sciences and BiotechnologyKorea UniversitySeoulKorea
  2. 2.Biomedical Research InstituteSeoul National University HospitalSeoulKorea

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