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《Chinese Journal of Cellular and Molecular Immunology》 2012-05
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Biological characteristics of CD55~(hig) side population in breast cancer cell line MCF-7

YANG Chao1,CUI Zhi-chao2,LIU Yun-jiang1,WANG Shi-jie3 1Department of Breast Center,the Fourth Hospital,Hebei Medical University,Shijiazhuang 050011;2Department of the First Mammary,the People' s Hospital of Tangshan,Tangshan 063000;3Department of Endocrinology,the Fourth Hospital,Hebei Medical University,Shijiazhuang 050011,China  
AIM: To determine whether the CD55hig expression(CD55hig) cells in side population(SP) of the cell line MCF-7 possess characteristics of cancer stem cells.METHODS: Breast cancer cell line MCF-7 was cultured and the nucleic acid dye Hoechst33342 and Verapami were added.Flow cytometry(FCM) was employed to isolate the cells of SP and mian population(MP),the cells were then labeled with CD55 mAb;mean values of fluorescence intensity of CD55 in SP and MP cells were measured.CD55 mAb was used to mark the unsorted MCF-7 cells,the proportion of CD55hig cells was determined,and then CD55hig cells were sorted and collected,to observe biological characteristics,such as cell morphology,adherence rate,colony formation and cell cycle distribution as well as nude mice implantation.RESULTS: Mean value of fluorescence intensity of CD55 in SP cells was 100.85±4.57,and mean value of fluorescence intensity of CD55 in MP cells was 50.51±4.75;the proportion of CD55hig cell in MCF-7 cells was 2.12%;adherent rate of CD55hig cells in 24 h was lower than that in CD55low cells and 24 h after inoculation adherent rates of both cells had no significant difference(P0.05);CD55hig cells were mostly spherical and kept in a suspended state at 12 hours after inoculation and culture;CD55hig cells had the ability of colony formation,the clone-forming rate of CD55hig cells was(20.04±1.07)% when the cells were cultured for one week,it was lower than the rate of(27.14±1.07)% in CD55low cells(P0.01);the ratio of G0/G1 resting cells in CD55hig cells was(85.4±3.37)% which was higher than that in CD55low cells(58.6±2.55)% and in MCF-7 cells(70.73±4.21)%,which had a statistically significant difference(P0.05).All nude mice implanted with CD55hig cells developed the tumor,and the pathological examination of the transplanted tumor had the properties of malignant cells.CONCLUSION: A few CD55hig cells were found in breast cancer MCF-7 cells,and most of CD55hig cells were quiescent,non-adherent as well as being in a suspended state and being hereditary after cloning,so CD55hig cells had biological characteristics of cancer stem cells.
【Fund】: 河北省强势特色学科(2005);; 河北省自然基金(C2009001210)
【CateGory Index】: R737.9
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1 YANG Chao;CUI Zhichao;GUO Qingjun;LIU Yunjiang;WANG Shijie;Department of Breast Center,the Fourth Hospital,Hebei Medical University;Department of the First mammary,the People ’s Hospital of Tang Shan;Department of Educational Administration,Hebei Medical University;Department of Endoscope,the Fourth Hospital,Hebei Medical University;;The activity of special CTLs in diminishing CD55~(high) subgroup in human breast cancer MCF-7 cells[J];Chinese Journal of Cellular and Molecular Immunology;2013-11
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