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目的:研究不同甲状腺肿瘤细胞中是否存在具备干细胞特性的侧群(side population,SP)细胞,同时比较侧群细胞和非侧群(non-SP)细胞生长、成瘤及侵袭性的差异。方法:以多种人甲状腺未分化癌细胞株为对象,利用干细胞高表达ABCG2转运体,可将荧光染料Hoechst泵出胞外的特性,运用双波长流式细胞仪从不同的甲状腺肿瘤细胞株中分选出具有干细胞特性的侧群细胞,进而通过半定量PCR及免疫荧光染色比较侧群和非侧群细胞的干细胞标志OCT4及肿瘤耐药基因ABCG2、MDR1表达的情况,采用克隆形成率及细胞侵袭迁移实验比较两组细胞群的克隆形成能力和侵袭转移能力。结果:人甲状腺未分化癌细胞株中存在具有干细胞特性的侧群细胞,该群细胞高表达干细胞标志OCT4及肿瘤耐药基因ABCG2和MDR1;同时在体外培养中,此侧群细胞可产生侧群细胞和非侧群细胞;侧群细胞比非侧群细胞具有更强的体外克隆形成能力和侵袭转移能力。结论:甲状腺肿瘤细胞中存在具有干细胞特性的侧群细胞,此群细胞可生成普通的瘤体细胞,提示该群细胞可能是肿瘤耐药和复发的根源。
OBJECTIVE: To investigate the presence or absence of stem cell-specific side population (SP) cells in different thyroid tumor cells and to compare the growth, tumorigenicity and invasiveness of side-cell and non-SP cells. Methods: A variety of human undifferentiated thyroid cancer cell lines as an object, the use of high expression of ABCG2 stem cell transporter, the fluorescence dye Hoechst pump extracellular characteristics of the use of dual-wavelength flow cytometry from different thyroid tumor cell lines Colony-forming cells with stem cell characteristics were sorted out. Then, the expression of OCT4 and ABCG2 and MDR1 were compared by semi-quantitative PCR and immunofluorescence staining. Invasion and migration experiments were compared between the two groups of cell cloning ability and invasion and metastasis. RESULTS: Side population cells with stem cell characteristics existed in human thyroid undifferentiated carcinoma cell lines, which highly expressed stem cell marker OCT4 and tumor drug resistance genes ABCG2 and MDR1. At the same time, in vitro culture, side population cells could produce side population Cells and non-lateral cells; lateral cells than non-lateral cells have a stronger ability in vitro cloning ability and invasion and metastasis. CONCLUSIONS: Thyroid tumor cells present side population cells with stem cell characteristics, which can generate normal tumor cells, suggesting that the group of cells may be the source of tumor resistance and relapse.