CD44作为筛选下咽癌肿瘤干细胞分子标志物的价值  

CD44 as a molecular marker to screen tumor stem cells in hypopharyngeal carcinoma

作　　者：沈晨凌[1] 向明亮[1] 聂琛 胡海霞 马衍[1] 吴皓[1] 

机构地区：[1]上海交通大学医学院附属新华医院耳鼻咽喉头颈外科,上海交通大学医学院耳科学研究所,上海200092

出　　处：《中国耳鼻咽喉颅底外科杂志》2013年第5期388-392,398,共6页Chinese Journal of Otorhinolaryngology-skull Base Surgery

基　　金：国家自然科学基金资助项目(81271088);上海市自然科学基金资助项目(11ZR1423600)

摘　　要：目的探讨CD44作为筛选下咽癌肿瘤干细胞分子标记物的价值。方法应用流式细胞仪检测下咽癌FADU细胞系中CD44的表达;采用免疫磁珠分选技术(MACS)分选CD44+细胞,并用流式细胞仪检测其纯度;运用四甲基偶氮唑蓝(MTT)比色法检测CD44+、CD44-细胞的体外增殖能力,将相同数量级(1×106、1×105)的CD44+、CD44-细胞分别注射于雄性非肥胖糖尿病型/重症联合免疫缺陷(non-obese diabetic/severe combined immunodefi ciency,NOD/SCID)小鼠皮下,观察其体内致瘤能力的差异。结果下咽癌FADU细胞系中CD44+细胞占(21.1±1.56)%,免疫磁珠分选后的CD44+细胞纯度达(99.4±0.29)%。体外增殖能力研究显示CD44+细胞增殖速度明显快于CD44-细胞。NOD/SCID鼠体内致瘤实验结果显示:1×105CD44+细胞致瘤率为12.5%(1/8),1×105 CD4 4-细胞致瘤率为0(0/8);1×106 CD44+细胞致瘤率为100%(8/8),1×106 CD4 4-细胞致瘤率为5 0%(4/8);相同数量级的CD44+与CD44-两组细胞致瘤率差异具有统计学意义(P<0.05)。此外,CD44+细胞组比CD44-细胞组更早成瘤,潜伏期明显缩短。CD44+细胞组所致肿瘤体积平均为(2017.81±538.50)mm3,而CD44-细胞组所致肿瘤体积平均为(1153.25±503.18)mm3,两组差异具有统计学意义(t=2.67,P<0.05)。结论下咽癌肿瘤中CD44+细胞比CD44-细胞具有更强的体外增殖能力和体内致瘤能力,提示肿瘤干细胞可能存在于CD44+肿瘤细胞内,CD44可作为筛选下咽癌肿瘤干细胞的一个重要分子标记物。Objective To study the significance of CD44 as a molecular marker for screening tumor stem cells in hypopharyngeal carcinoma. Methods The CD44 expression levels in the hypopharyngeal carcinoma cell line FaDu were analyzed using flow eytometry. The CD44 ＋ and CD44-eell populations were sorted by using magnetic-associated cell sorting （ MACS ） technology. After separation, the purity of the CD44 ＋ cells was detected using flow eytometry. The MTT method was used to detect the different proliferation capability of the CD44 ＋ and CD44-cells in vitro. The tumorigenicity of the CD44 ＋ and CD44-cells was determined by injecting CD44 ＋ or CD44-cells （ 1 × 10^6 and 1 × 10^5 ） into the body of NOD/SCID mice. Results CD44 expressed in （21. 1±1.56）% of the FaDu cells. The CD44 ＋ population was efficiently sorted by MACS with the purity of （ 99.4 ± 0.29 ） %. The MTT assay indicated that the sorted CD44 ＋ cells had a stronger proliferative capacity than the CD44-eells. The tumorigenicity study showed that all the mice injected with 1 × 10^6 CD44 ＋ cells developed tumors （8/8） , half of the mice injected with 1 × 10^6 CD44-cells developed tumors （4/8）, 1 of the 8 mice injected with 1×10^5 CD44 ＋ cells developed tumors （12.5%）, but none of the mice injected with 1 ×10^5 CD44-cells developed any tumors （0/8）. At the same concentration, the difference in tumorigenic rates between the CD44 ＋ and CD44-groups was statistically significant （Fisher＇ s exact test, P 〈 0.05）. In addition, the CIM4 ＋ group had a shorter incubation period than the CD44- group. Furthermore, the average tumor volume of the CD44 ＋ group was （2017.81 ± 538.50） mm3, while that of the CD44-group was （ 1153.25±503. 18 ） mm3, and the difference was statistically significant （ t = 2. 67, P 〈 0.05 ）. Conclusions The CD44 ＋ cells have stronger proliferative capacity and higher tumorigenic potential than the CD44-cells, which suggests that the tumor stem cells of hypopharyngeal ca

关 键 词：肿瘤干细胞 分子标记物 CD44 下咽癌 

分 类 号：R34[医药卫生—基础医学] R739.91