摘要
目的:确定MCF-7细胞系侧群(SP)部分的CD55高表达(CD55hig)细胞是否具有肿瘤干细胞特性。方法:培养乳腺癌MCF-7细胞系,加入核酸染料Hoechst33342和Verapami,用流式细胞术(FCM)分离SP亚群和MP亚群细胞,以CD55mAb分别标记,测定侧群(SP)和主群(MP)细胞的CD55平均荧光强度值,再以CD55 mAb标记未分选MCF-7细胞,测定CD55hig细胞所占比例,并分选收集CD55hig细胞,观察细胞形态、贴壁率、克隆形成及细胞周期分布以及裸鼠体内种植等生物学特性。结果:SP细胞CD55的平均荧光强度值为100.85±4.57,MP细胞CD55的平均荧光值为50.51±4.75;MCF-7细胞中CD55hig细胞比例为2.12%;24 h内CD55hig细胞细胞贴壁率低于CD55low细胞,接种24 h后,两者贴壁率无明显差异(P>0.05);接种培养12 h后,CD55hig细胞多为球形呈悬浮状态;CD55hig细胞都具有克隆形成能力,CD55hig细胞在培养1周后克隆形成率为(20.04±1.07)%,低于CD55low细胞(27.14±1.07)%(P<0.01);CD55hig细胞中G0/G1静止期细胞占(85.4±3.37)%,高于CD55low细胞(58.6±2.55)%及MCF-7细胞(70.73±4.21)%,有统计学显著差异(P<0.05)。所有裸鼠种植CD55hig细胞后均成瘤,移植瘤病理检验具有恶性肿瘤细胞的特性。结论:乳腺癌MCF-7细胞中存在少量的CD55hig细胞,CD55hig细胞大多数处于静止期、不贴壁呈悬浮状态和克隆遗传性,CD55hig细胞具有肿瘤干细胞生物学特性。
AIM： To determine whether the CD55^hig expression （CD55^hig） 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 add- ed. 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 inten- sity of CD55 in SP and MP cells were measured. CD55 mAb was used to mark the unsorted MCF-7 cells, the proportion of CD55^hig cells was determined, and then CD55^hig cells were sorted and collected, to observe biological characteris- tics, such as cell morphology, adherence rate, colony for- mation 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 flu- orescence intensity of CD55 in MP cells was 50.51 ± 4.75 ; the proportion of 55^hig cell in MCF-7 cells was 2.12% ; ad- herent rate of CD55^hig cells in 24 h was lower than that in CD55^hig cells and 24 h after inoculation adherent rates of both cells had no significant difference （P 〉 0.05）; CD55^higcells were mostly spherical and kept in a suspended state at 12 hours after inoculation and culture; CD55^hig cells had the ability of colony formation, the clone-forming rate of CD55^hig 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 CD55^hig cells （P〈0.0]） ; the ratio of G0/G] resting cells in CD55^hig cells was （85.4 ± 3.37）% which was higher than that in CD55^hig w cells （58.6± 2.55 ） % and in MCF-7 cells （70. 73±4.21 ） %, which had a statistically significant differ ence （P〈0.05）. All nude mice implanted with CD55^hig cells developed the tumor, and the pathological examination of the transplanted tumor had the properties of malignant cells. CONCLUSION： A few CD55^higcells were found
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2012年第5期492-496,共5页
Chinese Journal of Cellular and Molecular Immunology
基金
河北省强势特色学科(2005)
河北省自然基金(C2009001210)
