中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (6): 1070-1074.doi: 10.3969/j.issn.1673-8225.2011.06.026

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

人淋巴瘤Raji细胞系中的边缘细胞:淋巴瘤干细胞存在之可能

谢  健1,李晓进2,金从国3,喻美佳2,李惠民1   

  1. 1昆明医学院第一附属医院血液科,云南省昆明市   650021
    2昆明医学院第四附属医院血液科,云南省昆明市 650031
    3昆明医学院第三附属医院肿瘤所,云南省昆明市  650106
  • 收稿日期:2010-07-07 修回日期:2010-10-18 出版日期:2011-02-05 发布日期:2011-02-05
  • 通讯作者: 李惠民,教授,主任医师,昆明医学院第一附属医院血液科,云南省昆明市 650021 lihuimin@medmail.com.cn
  • 作者简介:谢健★,男,1982年生,四川省乐山市人,汉族,2008年昆明医学院毕业,硕士,医师,现工作于四川省宜宾市第一人民医院血液科,主要从事恶性血液病的诊治。 ls_xiejian@163.com
  • 基金资助:

    云南省自然科学基金资助项目(2006C0076M)。课题名称:白血病细胞株边缘细胞分离、鉴定及生物学特性研究。

Side population cells in human lymphoma Raji cell line: Possibility of lymphoma stem cells

Xie Jian1, Li Xiao-jin2, Jin Cong-guo3, Yu Mei-jia2, Li Hui-min1   

  1. 1Department of Hematology, First Affiliated Hospital, Kunming Medical University, Kunming  650021, Yunnan Province, China
    2Department of Hematology, Fourth Affiliated Hospital, Kunming Medical University, Kunming  650031, Yunnan Province, China
    3Institute of Tumor, Third Affiliated Hospital, Kunming Medical University, Kunming  650106, Yunnan Province, China
  • Received:2010-07-07 Revised:2010-10-18 Online:2011-02-05 Published:2011-02-05
  • Contact: Li Hui-min, Professor, Chief physician, Department of Hematology, First Affiliated Hospital, Kunming Medical University, Kunming 650021, Yunnan Province, China lihuimin@medmail.com.cn
  • About author:Xie Jian★, Master, Physician, Department of Hematology, First Affiliated Hospital, Kunming Medical University, Kunming 650021, Yunnan Province, China ls_xiejian@163.com
  • Supported by:

    the Natural Science Foundation of Yunnan Province, No. 2006C0076M*

摘要:

背景:已有研究提示急性白血病、多发性骨髓瘤中肿瘤干细胞富集于SP细胞中,且免疫表型及生物学特性相比于主群细胞存在明显的异质性。而关于淋巴瘤淋巴瘤干细胞的研究报道甚少。
目的:检测人淋巴瘤Raji细胞系中是否存在SP细胞,分选该群细胞后观察其与非SP细胞在分化抗原和P-糖蛋白表达以及细胞周期方面的差异,探讨淋巴瘤干细胞存在的可能性。
方法:Hoechst-33342染色细胞后运用带紫外激发光的流式细胞仪进行Raji细胞系SP细胞的检测及分选。观察不同质量浓度维拉帕米对SP细胞抑制的程度,选择最佳抑制浓度。分选后分别检测SP和非SP细胞P-糖蛋白、分化抗原CD34,CD19,CD20,CD5表达以及细胞周期,并进行对比。
结果与结论:Raji细胞系中存在SP细胞,比例在2%左右,当维拉帕米质量浓度为50 mg/L时能被完全抑制。SP和非SP细胞CD20阳性表达率分别为18.2%和93.6%,CD5阳性表达率分别为78.0%和22.2%;P-糖蛋白表达差异无显著性意义,但SP中阳性细胞的平均荧光强度强于非SP细胞。提示Raji细胞系中SP与非SP细胞在分化程度、多药耐药性方面具有明显的差异,SP细胞是具有明显“异质性”的细胞亚群。据此可初步推测,Raji细胞系的构成也类似肿瘤干细胞分级组成模式,而淋巴瘤干细胞则有可能富集于SP亚群中。

关键词: 淋巴瘤, SP细胞, 异质性, 分化抗原, 肿瘤干细胞

Abstract:

BACKGROUND: Previous studies have demonstrated that cancer stem cells enriched SP cells in the acute leukemia and multiple myeloma. Moreover, immunophenotype and biological characteristics have significant heterogeneity compared with main population cells. However, there is few study concerning lymphoma stem cells. 
OBJECTIVE: To examine whether the human lymphoma cell line-Raji contains SP cells, to investigate the difference in differentiation antigen, P-glycoprotein expression and cell cycle between SP cells and non-SP cells, and to explore the existing possibility of lymphoma stem cells.
METHODS: After Hoechst-33342 staining, Raji cell line SP cells were detected and sorted using flow cytometry with ultraviolet. Inhibitory effects of various mass concentrations of verapamil on SP cells were observed and an optimal inhibitory concentration was selected. After sorting, P-glycoprotein, differentiation antigen CD34, CD20, CD19, CD5 and cell cycle were determined and compared between SP and non-SP cells.
RESULTS AND CONCLUSION: SP cells accounted for about 2% of viable cells in Raji, and could be blocked when verapamil was 50 mg/L. SP expressed 18.2% CD20+ cells compared with non-SP (93.6% CD20+), 78.0% CD5+cells compared with non-SP (22.2% CD5+). No significant difference in P-glycoprotein expression was detected between the two populations, but the P-glycoprotein (+) cells in SP displayed more intensive fluorescence compared with non-SP. These indicated that there are significant differences in differentiation degree and multidrug resistance between SP cells and non-SP cells in Raji cell line. SP cells are heterogeneous subset. We supposed that Raji cell line might be organized similarly as cancer stem cell hierarchy, and it is possible that lymphoma stem cells are enriched in SP cells.

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