title: A novel approach for enriching cancer stem cells from the human SW-13 adrenocortical carcinoma cell line.
abstract: The present study was undertaken to develop a new method for enriching cancer stem cells (CSCs) from the human adrenal cortical carcinoma (ACC) cell line SW-13. Given that the existence of CSCs in ACC causes resistance to conventional chemotherapies, treatment with cyclophosphamide was used for in vivo selection of CSCs in a BALB/c nude mouse tumor xenograft model established using the ACC cell line SW-13. The characteristics of CSCs in three generations of tumor xenografts were assessed for single-cell colony formation, flat colony formation, and cell sphere formation in serum-free suspension culture. The formation rates of single-cell colonies, flat colonies, and cell spheres were significantly higher for tumor xenograft cells treated with cyclophosphamide than for untreated engrafted tumor cells. Flow cytometry to examine expression of the CSC markers C-X-C chemokine receptor type-4 (CXCR4; CD184) and ATP-binding cassette sub-family G member-2 (ABCG2; CDw338) revealed markedly higher levels of CXCR4 and ABCG2 in cyclophosphamide-treated xenograft tumor cells compared to untreated tumor cells. Together, these results indicate that cyclophosphamide treatment of tumor xenograft cells caused enrichment of CSCs with a strong capability for self-renewal and proliferation. In this method, the administration of cyclophosphamide selectively kills cancer cells without toxicity to CSCs and thereby provides a practical approach for achieving the enrichment of CSCs in ACC.
authors.affiliation: National Institutes of Health, Bethesda, MD 20892, U.S.A. E-mail: liquenti@mail.nih.gov or Dr. Zuojie Luo, Department of Endocrinology, First Hospital of Guangxi Medical University, Nanning 530021, China. zluo888@163.com.
authors.forename: Wenqing
authors.initials: W
authors.lastname: Zeng
authors.forename: Xiaozhou
authors.initials: X
authors.lastname: Chen
authors.forename: Yan
authors.initials: Y
authors.lastname: Ma
authors.forename: Zhenxing
authors.initials: Z
authors.lastname: Huang
authors.forename: Yuan
authors.initials: Y
authors.lastname: Qin
authors.forename: Fengping
authors.initials: F
authors.lastname: Wu
authors.forename: Lili
authors.initials: L
authors.lastname: Wu
authors.forename: Xinghuan
authors.initials: X
authors.lastname: Liang
authors.forename: Yingfen
authors.initials: Y
authors.lastname: Qin
authors.forename: Jia
authors.initials: J
authors.lastname: Zhou
authors.forename: Decheng
authors.initials: D
authors.lastname: Lu
authors.forename: Xiaocong
authors.initials: X
authors.lastname: Kuang
authors.forename: Qingdi Quentin
authors.initials: QQ
authors.lastname: Li
authors.forename: Zuojie
authors.initials: Z
authors.lastname: Luo
mesh_terms: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000306:Adrenal Cortex Neoplasms / Q000188:drug therapy / Q000473:pathology*; D018268:Adrenocortical Carcinoma / Q000188:drug therapy / Q000473:pathology*; D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology; D002469:Cell Separation / Q000379:methods*; D003520:Cyclophosphamide / Q000494:pharmacology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D019718:Receptors, CXCR4 / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay
chemical_list: D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; C501412:Abcg2 protein, mouse; D018906:Antineoplastic Agents, Alkylating; C514936:CXCR4 protein, mouse; D019718:Receptors, CXCR4; D003520:Cyclophosphamide
keywords: ABCG2; Adrenocortical carcinoma; CXCR4; cancer stem cell; chemotherapy; cyclophosphamide