Berberine Diminishes the Side Population and ABCG2 Transporter Expression in MCF-7 Breast Cancer Cells

Abstract

The plant alkaloid berberine has many biological activities including the ability to induce cell cycle arrest and apoptosis, making it a potentially useful agent for targeting cancer cells. We have analyzed the effects of berberine on MCF-7 breast cancer cells. Berberine was added to MCF-7 cells in culture, and proliferation, side population (SP) cells and expression of ABCG2 were examined. Berberine caused a dose-dependent reduction in proliferation. Hoechst 33 342 dye staining and FACS analysis revealed that berberine treatment caused a decrease in SP cells relative to untreated controls. In addition, berberine treatment was associated with a decrease in expression of ABCG2 relative to untreated controls. These results indicate that the growth inhibitory effects of berberine treatment on MCF-7 cells may be partly via effects on SP and ABCG2 expression. Further work is warranted to explore whether berberine may be a novel therapeutic drug useful for targeting breast cancer stem cells.

Key words

berberine - breast cancer - cancer stem cells - side population - ABCG2

References

• 1 Phillips T M, McBride W H, Pajonk F. The response of CD24(-/low)/CD44+ breast cancer initiating cells to radiation.  J Natl Cancer Inst. 2006;  98 1777-85
• 2 Burkert J, Otto W, Wright N. Side populations of gastrointestinal cancers are not enriched in stem cells.  J Pathol. 2008;  214 564-73
• 3 Ho M M, Ng A V, Lam S, Hung J Y. Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells.  Cancer Res. 2007;  67 4827-33
• 4 Scharenberg C W, Harkey M A, Torok-Storb B. The ABCG2 transporter is an efficient Hoechst 33 342 efflux pump and is preferentially expressed by immature human hematopoietic progenitors.  Blood. 2002;  99 507-12
• 5 Patrawala L, Calhoun T, Schneider-Broussard R, Zhou J, Claypool K, Tang D G. Side population is enriched in tumorigenic, stem-like cancer cells, whereas ABCG2+ and ABCG2- cancer cells are similarly tumorigenic.  Cancer Res. 2005;  65 6207-19
• 6 Lee S, Lim H J, Park H Y, Lee K S, Park J H, Jang Y. Berberine inhibits rat vascular smooth muscle cell proliferation and migration in vitro and improves neointima formation after balloon injury in vivo. Berberine improves neointima formation in a rat model.  Artherosclerosis. 2006;  186 29-37
• 7 Jantova S, Cipak L, Cernakova M, Kost'alova D. Effect of berberine on proliferation, cell cycle and apoptosis in HeLa and L1210 cells.  J Pharm Pharmacol. 2003;  55 1143-9
• 8 Leng S H, Lu F E, Xu L J. Therapeutic effects of berberine in impaired glucose tolerance rats and its influence on insulin secretion.  Acta Pharmacol Sin. 2004;  25 496-502
• 9 Abidi P, Zhou Y, Jiang J D, Liu J. Extracellular signal-regulated kinase-dependent stabilization of hepatic low-density lipoprotein receptor mRNA by herbal medicine berberine.  Arterioscler Thromb Vasc Biol. 2005;  25 2170-6
• 10 Mantena S K, Sharma S D, Katiyar S K. Berberine inhibits growth, induces G1 arrest and apoptosis in human epidermoid carcinoma A431 cells by regulating Cdki-Cdk-cyclin cascade, disruption of mitochondrial membrane potential and cleavage of caspase 3 and PARP.  Carcinogenesis. 2006;  27 2018-27
• 11 Letasiova S, Jantova S, Miko M, Ovadekova R, Horvathova M. Effect of berberine on proliferation, biosynthesis of macromolecules, cell cycle and induction of intercalation with DNA, dsDNA damage and apoptosis in Ehrlich ascites carcinoma cells.  J Pharm Pharmacol. 2006;  58 263-70
• 12 Orfila L, Rodriguez M, Colman T, Hasegawa M, Merentes E, Arvelo F. Structural modification of berberine alkaloids in relation to cytotoxic activity in vitro. .  J Ethnopharmacol. 2000;  71 449-56
• 13 Kim J B, Lee K M, Ko E, Han W, Lee J E, Shin I. et al . Berberine inhibits growth of the breast cancer cell lines MCF-7 and MDA-MB-231.  Planta Med. 2008;  74 39-42
• 14 Yin L, Castagnino P, Assoian R K. ABCG2 expression and side population abundance regulated by a transforming growth factor beta-directed epithelial-mesenchymal transition.  Cancer Res. 2008;  68 800-7
• 15 Goodell M A. Multipotential stem cells and ”side population” cells.  Cytotherapy. 2002;  4 507-8
• 16 Szotek P P, Pieretti-Vanmarcke R, Masiakos P T, Dinulescu D M, Connolly D, Foster R. et al . Ovarian cancer side population defines cells with stem cell-like characteristics and Mullerian inhibiting substance responsiveness.  Proc Natl Acad Sci U S A. 2006;  103 11 154-9
• 17 Wu C, Wei Q, Utomo V, Nadesan P, Whetstone H, Kandel R. et al . Side population cells isolated from mesenchymal neoplasms have tumor initiating potential.  Cancer Res. 2007;  67 8216-22
• 18 Zhou J, Wulfkuhle J, Zhang H, Gu P, Yang Y, Deng J. et al . Activation of the PTEN/mTOR/STAT3 pathway in breast cancer stem-like cells is required for viability and maintenance.  Proc Natl Acad Sci U S A. 2007;  104 16 158-63
• 19 Tang Y, Kitisin K, Jogunoori W, Li C, Deng C X, Mueller S C. et al . Progenitor/stem cells give rise to liver cancer due to aberrant TGF-beta and IL-6 signaling.  Proc Natl Acad Sci U S A. 2008;  105 2445-50
• 20 Loontiens F G, Regenfuss P, Zechel A, Dumortier L, Clegg R M. Binding characteristics of Hoechst 33 258 with calf thymus DNA, poly[d(A-T)], and d(CCGGAATTCCGG): multiple stoichiometries and determination of tight binding with a wide spectrum of site affinities.  Biochemistry. 1990;  29 9029-39
• 21 Arndt-Jovin D J, Jovin T M. Analysis and sorting of living cells according to deoxyribonucleic acid content.  J Histochem Cytochem. 1977;  25 585-9
• 22 Gregoire M, Hernandez-Verdun D, Bouteille M. Visualization of chromatin distribution in living PTO cells by Hoechst 33 342 fluorescent staining.  Exp Cell Res. 1984;  152 38-46
• 23 Kang M R, Chung I K. Down-regulation of DNA topoisomerase IIalpha in human colorectal carcinoma cells resistant to a protoberberine alkaloid, berberrubine.  Mol Pharmacol. 2002;  61 879-84
• 24 Kim S A, Kwon Y, Kim J H, Muller M T, Chung I K. Induction of topoisomerase II-mediated DNA cleavage by a protoberberine alkaloid, berberrubine.  Biochemistry. 1998;  37 16 316-24
• 25 Scharff O, Foder B. Effect of trifluoperazine, compound 48/80, TMB-8 and verapamil on the rate of calmodulin binding to erythrocyte Ca²⁺-ATPase.  Biochim Biophys Acta. 1984;  772 29-36
• 26 Wu S N, Yu H S, Jan C R, Li H F, Yu C L. Inhibitory effects of berberine on voltage- and calcium-activated potassium currents in human myeloma cells.  Life Sci. 1998;  62 2283-94
• 27 Shitan N, Tanaka M, Terai K, Ueda K, Yazaki K. Human MDR1 and MRP1 recognize berberine as their transport substrate.  Biosci Biotechnol Biochem. 2007;  71 242-5
• 28 Kim M, Turnquist H, Jackson J, Sgagias M, Yan Y, Gong M. et al . The multidrug resistance transporter ABCG2 (breast cancer resistance protein 1) effluxes Hoechst 33 342 and is overexpressed in hematopoietic stem cells.  Clin Cancer Res. 2002;  8 22-8
• 29 Zhou S, Schuetz J D, Bunting K D, Colapietro A M, Sampath J, Morris J J. et al . 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. 2001;  7 1028-34
• 30 Christgen M, Ballmaier M, Bruchhardt H, von Wasielewski R, Kreipe H, Lehmann U. Identification of a distinct side population of cancer cells in the Cal-51 human breast carcinoma cell line.  Mol Cell Biochem. 2007;  306 201-12