title: Bufalin inhibits the differentiation and proliferation of human osteosarcoma cell line hMG63-derived cancer stem cells.
abstract: Cancer stem cells (CSCs) play an important role in drug resistance of tumor and are responsible for high recurrence rates. Agents that can suppress the proliferation and differentiation of CSCs would provide new opportunity to fight against tumor recurrence. In this study, we developed a new strategy to enrich CSCs in human osteosarcoma cell line hMG63. Using these CSCs as model, we tested the effect of bufalin, a traditional Chinese medicine, on the proliferation and differentiation of CSCs. hMG63 cells were cultured in poly-HEMA-treated dish and cancer stem cell-specific medium. In this nonadhesive culture system, hMG63 formed spheres, which were then collected and injected into the immunodeficient mice. Cisplatin was administered every 3 days for five times. The enriched xenograft tumors were cultured in cancer stem cell-specific medium again to form tumor spheres. Expression of cancer stem cell markers of these cells was measured by flow cytometry. These cells were then treated with bufalin, and the proliferation and differentiation ability were indicated by the expression level of molecular markers and the formation of sphere again in vitro. We obtained a low CD133+/CD44 cell population with high-level stem cell marker. When treated with bufalin, the sphere could not get attached to the flask and failed to differentiate, which was indicated by the stable expression of stem cell marker CD133 and OCT-4 in the condition permissive to differentiation. Treatment of bufalin also suppressed the single cells isolated from the sphere to form sphere again in the nonadhesive culture system, and a decreased expression of proliferation marker Ki67 was also detected in these cells. Sphere-formed and chemoresistant colon xenograft tumors in immunodeficient mice could enrich cancer stem cell population. Bufalin could inhibit proliferation and differentiation of CSCs.
authors.affiliation: Department of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Room 1002, Road Yinghua, Shanghai, China, cyw188@yeah.net.
authors.forename: Yuewen
authors.initials: Y
authors.lastname: Chang
authors.forename: Yongfang
authors.initials: Y
authors.lastname: Zhao
authors.forename: Hongsheng
authors.initials: H
authors.lastname: Zhan
authors.forename: Xiaoen
authors.initials: X
authors.lastname: Wei
authors.forename: Tianjin
authors.initials: T
authors.lastname: Liu
authors.forename: Bo
authors.initials: B
authors.lastname: Zheng
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000235:genetics; D002018:Bufanolides / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002945:Cisplatin / Q000008:administration & dosage; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic; D006023:Glycoproteins / Q000235:genetics; D006801:Humans; D051379:Mice; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000473:pathology; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D012516:Osteosarcoma / Q000235:genetics* / Q000378:metabolism; D010455:Peptides / Q000235:genetics
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D002018:Bufanolides; D006023:Glycoproteins; D050814:Octamer Transcription Factor-3; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D002945:Cisplatin; C022777:bufalin