结肠癌干细胞的肿瘤形成特征及其临床价值  

Tumorigenic features of colon cancer stem cells

作　　者：邓超[1] 陈豪[1] 金黑鹰[2,3] 王水明[2,3] 

机构地区：[1]南京中医药大学第一临床学院,江苏省南京市210068 [2]南京中医药大学第三附属医院 [3]南京市中医院肛肠科六病区 全国肛肠医疗中心 江苏省中西医结合结直肠癌诊疗中心,江苏省南京市210001

出　　处：《世界华人消化杂志》2015年第3期363-372,共10页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;Nos.30973837;81273944~~

摘　　要：目的:研究结肠癌干细胞的肿瘤形成价值及其可能的临床价值.方法:对结肠癌干细胞进行无血清培养,以流式细胞仪检测CD133和CD44表达,以5-氟尿嘧啶(5-flurouracil,5-Fu)处理结肠癌干细胞并观察其增殖抑制,对结肠癌干细胞进行动物实验以确定其成瘤特征.结果:Hoechest33342染色后,HT-29干细胞相对于HT-29细胞有明显的核拒染现象;流式细胞仪发现HT-29细胞CD133阳性表达比率为44.6%,C D44阳性表达比率为0.6%;而经过培养提纯后的HT-29干细胞CD133阳性表达比率为92.6%,CD44阳性表达比率为97.8%,C D 1 3 3与C D 4 4阳性表达比率显著提高;HT-29干细胞相对于HT-29细胞对5-Fu有明显的耐药作用,HT-29细胞IC50值为1.394μg/m L,HT-29干细胞的IC50值为13.087μg/m L,IC50值有明显的增高在恢复血清培养后,体外增殖活性明显增强;HT-29干细胞体内生长速度比H T-29细胞体内生长速度显著要缓慢,H T-29细胞的肿瘤体积是HT-29干细胞肿瘤体积的两倍多;HT-29干细胞在小鼠富血管区(腋下和腹股沟淋巴结)的生长速率比皮下生长速率有明显的提高,但与HT-29细胞的生长速率还有明显差距.结论:在结肠癌细胞株存在结肠癌干细胞,通过无血清培养能分离出具有干细胞特征的结肠癌干细胞株,结肠癌干细胞对5-Fu具有耐药性,在适当微环境下其能快速生长,转化为成熟的结肠癌细胞,提示了改变结肠癌干细胞微环境可能是防止肿瘤复发的一个途径.AIM： To explore the tumorigenic features of colon cancer stem cells and to analyze their clinical significance. METHODS： HT-29 colon cancer cells were cultured in serum free medium,and t h e expression of CD133 and CD44 was detected by flow cytometry. The inhibitory effect of 5-flurouracil（5-Fu） on HT-29 cells and HT-29 cancer stem cells was assessed, and the tumorigenic features of HT-29 cancer stem cells were studied using a mouse model. RESULTS： Hoechest33342 staining showed that less intense nuclear staining was observed in HT-29 cancer stem cells compared with HT-29 cancer cells. The positive expression rates of CD133 and CD44 were 44.6% and 0.6% in HT-29 cells, and 92.6% and 97.8% in HT-29 cancer stem cells, respectively. HT-29 cancer stem cells were resistant to 5-Fu and the half maximal（50%） inhibitory concentration（IC） of 5-Fu was 13.087 μg/m L, significantly higher than that for HT-29 cells（1.394 μg/m L）. In mouse models, the growth rate of HT-29 cancer stem cells was slower than that of HT-29 cells, but the growth rate of HT-29 cancer stem cells in lymph nodes was more quickly than that under the skin. CONCLUSION： There is a small proportion of cancer stem cells among HT-29 colon cancer cell line. Colon cancer stem cell can be isolated by culture in serum-free medium, and the stem cells show 5-Fu resistance. Colon cancer stem cells can progress to mature cancer cells in a proper microenvironment, which implies that changing the microenvironments of cancer stem cells can prevent the cancer stem cells from progressing to mature cancer cells.

关 键 词：结肠癌干细胞 分离 成瘤特征 临床意义 

分 类 号：R735.35[医药卫生—肿瘤]