| 摘要: |
| 目的:应用p75NTR进行人食管肿瘤干细胞的分选,并对其生物学特性进行鉴定。方法:对人食管癌标本癌细胞及食管癌细胞株TE-1、Eca109进行培养,应用流式细胞仪检测p75NTR在人食管癌细胞（esophageal cancer cells,ECCs）中的表达,应用磁珠分选（MACS）法进行p75NTR阳性细胞与阴性细胞的分选,观察p75NTR阳性细胞的增殖、分化及软琼脂克隆形成能力,并进行裸鼠接种以观察其致瘤能力；应用化疗药物作用于ECCs后检测其中p75NTR阳性细胞与阴性细胞存活率,以评价p75NTR阳性细胞对化疗药物的耐受性。结果:8个食管肿瘤细胞系（株）中,除SHEC-1、SHEC-5未检测到p75NTR阳性细胞外,其余6个细胞系（株）SHEC-4、SHEC-6、SHEC-7、SHEC-8、Eca109、TE-1中均检测到p75NTR阳性细胞,阳性细胞比例分别为 2.71%、0.32%、3.35%、1.13%、2.15%、0.45%。与阴性及未分选细胞相比,MACS分选后的p75NTR阳性细胞具有较强的增殖能力,具有分化产生其他表型细胞的能力,在软琼脂中具有较强的克隆形成能力(P<0.01)；行裸鼠接种时,p75NTR阳性细胞表现出较强的致瘤性,其中SHEC-7细胞只需2×103个即可致瘤,其致瘤能力是未分选细胞的50倍。化疗药物分别作用于p75NTR阳性细胞与阴性细胞48 h后,p75NTR阳性细胞的存活比例明显高于阴性细胞(P<0.05)。结论:人食管肿瘤细胞中p75NTR阳性细胞具有自我更新、分化、增殖能力,对化疗药物具有较强的耐受性,并具有较强的致瘤能力,具有肿瘤干细胞的特性。 |
| 关键词: 食管肿瘤 肿瘤干细胞 p75NTR 细胞分选 致癌性试验 肿瘤抗药性 |
| DOI：10.3724/SP.J.1008.2009.0481 |
| 投稿时间：2008-03-03修订日期：2009-03-26 |
| 基金项目:国家自然科学基金（30471718）. |
|
| p75NTR in isolation and identification of cancer stem cells from human esophageal carcinoma |
| SUN Zhi-gang,HUANG Sheng-dong*,ZHANG Bao-ren,XU Zhi-yun,LIU Xiao-hong,GONG De-jun,YUAN Yang |
| (Department of Cardiothoracic Surgery,Changhai Hospital,Second Military Medical University,Institute of Cardiothoracic) |
| Abstract: |
| Objective:To isolate and identify cancer stem cells from esophageal cancer cells (ECCs) using cell surface marker p75NTR.Methods: ECCs were cultured from surgically resected ECC specimens; ECC cell lines TE-1 and Eca109 were also cultured.The expression of p75NTR in human ECCs was examined by flow cytometry.p75NTR positive cells were isolated from ECCs using magnetic activated cell sorting (MACS) method.The proliferation,differentiation,and the ability of colony-forming in soft agar of the p75NTR positive cells were observed.The p75NTR positive cells were injected into BALB/c nude mice subcutaneously to observe their tumorigenesis ability.The survival rates of p75NTR positive and negative cells were assessed after treated with chemotherapy drugs to evaluate the resistance of p75NTR positive cells.Results: Six out of the eight cell lines,including SHEC-4,SHEC-6,SHEC-7,SHEC-8,Eca109,and TE-1,were positive of p75NTR ,with the positive rates being 2.71%,0.32%,3.35%,1.13%,2.15%,and 0.45%,respectively.It was showed that p75NTR positive cells possessed higher proliferation ability compared with p75NTR negative cells (P＜0.01).The p75NTR positive cells had higher colony-forming ability in soft agar compared with p75NTR negative cells (P＜0.01).The p75NTR positive cells demonstrated stronger tumorigenesis ability in nude mice. As few as 2 000 SHEC-7 cells could give rise to new tumors in xenotransplantation,with a tumorigenesis ability 50 times as high as that of the p75NTR negative cells.When treated with chemotherapy drugs for 48 h,p75NTR positive cells had significantly higher survival rate than p75NTR negative cells (P<0.05).Conclusion: The p75NTR positive ECCs possess self-renewal,differentiation,and proliferation abilities; they are strongly resistant to chemotherapy drugs,which gives them strong tumorigenesis ability and the characters of tumor stem cells. |
| Key words: esophageal neoplasms tumor stem cells p75NTR cell storing tumorigenecity test neoplasm drug resistance |
