Biological characteristics of side population cells in multiple myeloma U266 cell line in relation to drug resistance

doi:10.3969/j.issn.2095-4344.2017.29.010

Abstract

Abstract:

BACKGROUND: Studies have shown that tumor stem cells are the basis and the main cause of tumor recurrence and chemotherapy failure. Therefore, the research on the drug resistance of tumor stem cells has become a hotspot in the field of stem cell research.
OBJECTIVE: To analyze the biological characteristics of side population (SP) cells in the U266 cell lines in relation to drug resistance.
METHODS: Flow cytometry was used to monitor the percentage of SP cells in the U266 cell lines. Fluorescence-activated cell sorter (FACS) analysis was used to isolate SP and non-SP (NSP) cells from the U266 cell lines. Further analyses of the cell cycle, multidrug resistant protein, methyl cellulose cloning assay of SP cells and NSP cells were performed. Real-time quantitative PCR was used to determine the expression of ABCG2 and MDR1 genes.
RESULTS AND CONCLUSION: There were (2.46±0.35)% SP cells in U266 cells, which were most in the G₀ phase. The ratio of G₀/G₁ in SP cells was (81.50±5.42)%, which was significantly higher than that in NSP cells [(39.85±3.21)%; P < 0.05]. The positive expression rates of P-gp and ABCG2 in SP cells were signficantly higher than those in NSP cells (P < 0.05). The cloning efficiency of SP cells was significantly higher than that of NSP cells (P < 0.05). The mRNA expression of ABCG2 and MDR1 was also significantly higher in SP cells than in NSP cells (P < 0.05). To conclude, a small subpopulation of the isolates of U266 cell lines belong to tumor stem cell-like SP cell subset, most of which are at the G₀ phase. ABCG2 and MDR1 genes are highly expressed in SP cells, which particularly plays a important role in the multidrug resistance of multiple myeloma stem cell lines.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Neoplastic Stem Cells, Multiple Myeloma, Multidrug Resistance-Associated Proteins, Tissue Engineering

CLC Number:

R394.2

Bao Qiu-ping, Li Hui-min, Li Xiao-jin, Yan Yan-hong. Biological characteristics of side population cells in multiple myeloma U266 cell line in relation to drug resistance[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(29): 4648-4653.

Figures/Tables 1

References

[1] Buehrer BM, Cheatham B. Isolation and characterization of human adipose-derived stem cells for use in tissue engineering. Methods Mol Biol. 2013;1001:1-11.

[2] Spath L, Rotilio V, Alessandrini M, et al. Explant-derived human dental pulp stem cells enhance differentiation and proliferation potentials. J Cell Mol Med. 2010;14(6B): 1635-1644.

[3] Matsui W, Huff CA, Wang Q, et al. Characterization of clonogenic multiple myeloma cells. Blood. 2004;103(6):2332- 2336.

[4] Huff CA, Matsui W. Multiple myeloma cancer stem cells. J Clin Oncol. 2008;26(17):2895-2900.

[5] Ghosh N, Matsui W. Cancer stem cells in multiple myeloma. Cancer Lett. 2009;277(1):1-7.

[6] Gottesman MM, Fojo T, Bates SE. Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer. 2002;2(1):48-58.

[7] Nguyen LV, Vanner R, Dirks P,et al. Cancer stem cells: an evolving concept.Nat Rev Cancer. 2012;12(2):133-143.

[8] Wang M, Wang Y, Zhong J. Side population cells and drug resistance in breast cancer. Mol Med Rep. 2015;11(6): 4297-4302.

[9] López J, Poitevin A, Mendoza-Martínez V, et al. Cancer-initiating cells derived from established cervical cell lines exhibit stem-cell markers and increased radioresistance. BMC Cancer. 2012;12:48.

[10] Harris MA, Yang H, Low BE, et al. Cancer stem cells are enriched in the side population cells in a mouse model of glioma. Cancer Res. 2008;68(24):10051-10059.

[11] Connell PP, Kron SJ, Weichselbaum RR. Relevance and irrelevance of DNA damage response to radiotherapy. DNA Repair (Amst). 2004;3(8-9):1245-1251.

[12] Jemal A, Bray F, Center MM, et al. Global cancer statistics. CA Cancer J Clin. 2011;61(2):69-90.

[13] Priya N, Sarcar S, Majumdar AS, et al. Explant culture: a simple, reproducible, efficient and economic technique for isolation of mesenchymal stromal cells from human adipose tissue and lipoaspirate. J Tissue Eng Regen Med. 2014;8(9): 706-716.

[14] Ishige I, Nagamura-Inoue T, Honda MJ, et al. Comparison of mesenchymal stem cells derived from arterial, venous, and Wharton's jelly explants of human umbilical cord. Int J Hematol. 2009;90(2):261-269.

[15] Miranville A, Heeschen C, Sengenès C, et al. Improvement of postnatal neovascularization by human adipose tissue-derived stem cells. Circulation. 2004;110(3):349-355.

[16] Yoshimura K, Shigeura T, Matsumoto D, et al. Characterization of freshly isolated and cultured cells derived from the fatty and fluid portions of liposuction aspirates. J Cell Physiol. 2006;208(1):64-76.

[17] Brzoska M, Geiger H, Gauer S, et al. Epithelial differentiation of human adipose tissue-derived adult stem cells. Biochem Biophys Res Commun. 2005;330(1):142-150.

[18] Sotiropoulou PA, Perez SA, Salagianni M, et al. Characterization of the optimal culture conditions for clinical scale production of human mesenchymal stem cells. Stem Cells. 2006;24(2):462-471.

[19] Ou Y, Yuan XD, Cai YN, et al. A novel ethanol-based method to induce differentiation of adipose-derived stromal cells into astrocytes. Neural Regen Res. 2011;6(10): 738-743.

[20] Baer PC, Griesche N, Luttmann W, et al. Human adipose-derived mesenchymal stem cells in vitro: evaluation of an optimal expansion medium preserving stemness. Cytotherapy. 2010;12(1):96-106.

[21] Campagnoli C, Roberts IA, Kumar S, et al. Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood. 2001;98(8): 2396-2402.

[22] Cai YN, Yuan XD, Ou Y, et al. Apoptosis during β-mercaptoethanol-induced differentiation of adult adipose-derived stromal cel ls into neurons. Neural Regen Res. 2011;6(10):750-755.

[23] Gronthos S, Franklin DM, Leddy HA, et al. Surface protein characterization of human adipose tissue-derived stromal cells. J Cell Physiol. 2001;189(1):54-63.

[24] Gu F, Gao JH, Lu F. Culture of the single cell clones of human adipose-derived stem cells and identification of their surface antigen expressions. Nan Fang Yi Ke Da Xue Xue Bao. 2008; 28(6):1067-1069.

[25] Rowlands AS, George PA, Cooper-White JJ. Directing osteogenic and myogenic differentiation of MSCs: interplay of stiffness and adhesive ligand presentation. Am J Physiol Cell Physiol. 2008;295(4):C1037-1044.

[26] Chong KY, Hsu CJ, Hung TH, et al. Wnt pathway activation and ABCB1 expression account for attenuation of proteasome inhibitor-mediated apoptosis in multidrug-resistant cancer cells.Cancer Biol Ther. 2015;16(1): 149-159.

[27] Tsubaki M, Komai M, Itoh T, et al. By inhibiting Src, verapamil and dasatinib overcome multidrug resistance via increased expression of Bim and decreased expressions of MDR1 and survivin in human multidrug-resistant myeloma cells. Leuk Res. 2014;38(1):121-130.

[28] Drain S, Flannely L, Drake MB, et al. Multidrug resistance gene expression and ABCB1 SNPs in plasma cell myeloma. Leuk Res. 2011;35(11):1457-1463.

[29] Kumar V, Varma N, Varma S, et al. Flow cytometric analysis of DNA indices, expression of p53 and multidrug resistance genes in multiple myeloma patients. Anal Quant Cytol Histol. 2004;26(5):271-277.

[30] Jian Z, Chen F, Xie Q, et al. Expression level of MDR1, and MRP in patients with malignant blood disease and its clinical significance. Hunan Yi Ke Da Xue Xue Bao. 1997;22(4): 344-346.

[31] Mimeault M, Batra SK. Functions of tumorigenic and migrating cancer progenitor cells in cancer progression and metastasis and their therapeutic implications. Cancer Metastasis Rev. 2007;26(1):203-214.

[32] Matsui W, Wang Q, Barber JP, et al. Clonogenic multiple myeloma progenitors, stem cell properties, and drug resistance. Cancer Res. 2008;68(1):190-197.

[33] Goodell MA, Brose K, Paradis G, et al. Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo. J Exp Med. 1996;183(4):1797-1806.

[34] Morrow CS, Peklak-Scott C, Bishwokarma B, et al. Multidrug resistance protein 1 (MRP1, ABCC1) mediates resistance to mitoxantrone via glutathione-dependent drug efflux. Mol Pharmacol. 2006;69(4):1499-1505.

[35] Al-Hajj M, Clarke MF. Self-renewal and solid tumor stem cells. Oncogene. 2004;23(43):7274-7282.

[36] Peacock CD, Wang Q, Gesell GS, et al. Hedgehog signaling maintains a tumor stem cell compartment in multiple myeloma. Proc Natl Acad Sci U S A. 2007;104(10):4048-4053.

[37] Clarke RB, Anderson E, Howell A, et al. Regulation of human breast epithelial stem cells. Cell Prolif. 2003;36 Suppl 1: 45-58.

[38] Domen J, Weissman IL. Hematopoietic stem cells need two signals to prevent apoptosis; BCL-2 can provide one of these, Kitl/c-Kit signaling the other. J Exp Med. 2000;192(12): 1707-1718.

[39] Wang K, Wu X, Wang J, et al. Cancer stem cell theory: therapeutic implications for nanomedicine. Int J Nanomedicine. 2013;8:899-908.

[40] Unno K, Jain M, Liao R. Cardiac side population cells: moving toward the center stage in cardiac regeneration. Circ Res. 2012;110(10):1355-1363.