原代胃癌干细胞CD44表达意义探讨  

Expression and significance of CD44 protein in primary gastric stem cells

作　　者：彭平[1] 汪蕾[1] 蒋继宗[1] 晁腾飞[1] 庄亮[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肿瘤中心,湖北武汉430030

出　　处：《中华肿瘤防治杂志》2017年第3期173-177,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81201820)

摘　　要：目的肿瘤干细胞(cancer stem cells,CSCs)在恶性肿瘤的发生发展中起着重要作用,且是肿瘤耐药及复发的根源。本研究探讨CSCs表面标记蛋白CD44在分离及鉴定胃癌干细胞中的重要作用。方法收集2015-11-06-2015-12-01华中科技大学同济医学院附属同济医院3例胃癌患者手术标本。首先采用胶原酶消化法获取胃癌细胞,部分细胞分别贴壁及悬浮培养,并采用免疫荧光及流式分析的方法检测胃癌球形体中CD44的表达情况。部分细胞采用流式细胞仪分选得到CD44^+及CD44^-胃癌细胞,并通过无血清悬浮培养法比较两群细胞的球形体形成能力。结果免疫荧光显示,CD44蛋白在球形体中高表达;流式分析发现,贴壁的原代胃癌细胞中CD44^+细胞的比例为(30.73±7.6)%,而球形体中CD44^+细胞的比例高达(56.4±5.58)%,差异有统计学意义,P=0.009。球形体形成实验结果表明,与CD44^-细胞相比,CD44^+胃癌细胞具有更强的球形体形成能力,1 000个CD44^+及CD44^-胃癌细胞分别形成(14.33±2.52)和(1.67±0.58)个球形体,差异有统计学意义,P=0.001。通过比较发现,球形体培养对胃癌干细胞的富集效率为0.3%,而CD44对胃癌干细胞的富集效率为1.4%。结论 CD44是胃癌干细胞的表面标记蛋白,其可能成为胃癌靶向治疗的可靠靶标。OBJECTIVE Cancer stem cell plays an important role in the incidence and development of malignant tumor,and it is the root of tumor drug resistance and relapse.The objective of this paper was to investigate the expression of cancer stem cell surface marker CD44 in gastric cancer and its clinical value.METHODS Primary gastric cancer cells were collected from three patients in Tongji Hospital between November 6,2015 and December 1,2015.Isolated tumor samples were disaggregated into single cells by collagenase digestion.Part of cells were cultured in adherent and suspension medium respectively,and the expression of CD44 protein in gastric cancer stem cells were tested by immunofluorescence and flow cytometry.Part of cells were sorted to CD44^＋and CD44^-cells by flow cytometry,then,the sphere formation capacity of the two populations were compared by the serum-free suspension culture method.RESULTS CD44 protein expressed higher in sphere by Immunofluorescence test.Compared with the attached gastric cancer cells,the proportion of CD44^＋cells in the spheroid cells was much higher[（30.73±7.6）% vs（56.4±5.58）%],the difference was statistically significant（n=3,P=0.009）.Compared with CD44^-cells,the sphere formation ability of CD44^＋cells were much more stronger,the number of sphere formed by 1 000CD44^＋cells were 14.33±2.52,while the number of sphere formed by CD44^-cells were only 1.67±0.58,the difference was statistically significant（n=3,P=0.001）.The enrichment efficiency of the gastric cancer stem cells was only 0.3% by sphere,while which was 1.4% by CD44^＋cells.CONCLUSION CD44 is the surface marker of gastric cancer stem cells,and it could become a reliable target for the gastric cancer targeted therapy.

关 键 词：胃癌 肿瘤干细胞 CD44 球形体 

分 类 号：R735.2[医药卫生—肿瘤]