肝细胞性肝癌组织CD133^+细胞肿瘤干细胞特性的研究  

Identification and characterization of cancer stem cells from CD133^+ cells separated from hepatocellular carcinoma tissue

作　　者：林家耀 张志明[1] 刘剑勇[1] 劳明[2] 唐凯[2] 季风[1] 高胜[1] 李长仔[1] 蔡娟[1] 吕庆杰[1] 

机构地区：[1]广西医科大学附属肿瘤医院肝胆外科,广西南宁530021 [2]广西壮族自治区肿瘤防治研究所检验科,广西南宁530021

出　　处：《中华肿瘤防治杂志》2016年第4期223-227,232,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：广西科学研究与技术开发计划(桂科攻1355005-3-3)

摘　　要：目的越来越多的研究表明,肝癌细胞系中能检测到肿瘤干细胞(cancer stem cells,CSCs)的存在。本研究旨在探讨从肝细胞性肝癌(hepatocellular carcinoma,HCC)组织样本中分离获得的CD133+细胞是否具有CSCs特性。方法将2014-02-01-2015-06-30广西医科大学附属肿瘤医院肝胆外科25例手术获得的新鲜HCC组织和对应癌旁组织,采用酶消化法分别消化成单个肝癌细胞和单个肝细胞,利用流式细胞术检测部分单个肝癌细胞和单个肝细胞CD133的表达率。用剩余的单个肝癌细胞进行原代培养,流式细胞术将培养获得的肝癌细胞分选为CD133+和CD133-细胞,通过平板克隆形成实验、肿瘤球形成实验和裸鼠移植瘤形成实验对比分析这两组细胞的CSCs特性。结果 25例HCC组织中CD133的表达率为3.8%~8.3%,平均值为(5.8±1.6)%,而癌旁组织CD133的表达率为0.1%~0.4%,平均值为(0.2±0.1)%,两者比较差异有统计学意义,t=17.12,P<0.001。CD133+和CD133-细胞的平均克隆率分别为(25.2±0.8)%和(7.6±0.8)%,两者比较差异有统计学意义,t=81.95,P<0.001。CD133+和CD133-细胞的平均成球率分别为(20.3±0.6)%和(12.5±1.4)%,两者比较差异有统计学意义,t=68.17,P<0.001。CD133+细胞的裸鼠移植瘤形成能力明显高于CD133-细胞。结论从HCC组织样本中分离获得的CD133+细胞具有明显的CSCs特性。OBJECTIVE Accumulating evidences have shown that cancer stem cells（CSCs） exist in hepatocellμLar carcinoma（HCC） cell line. This study was to investigate whether CD133＋ cells separated from HCC tissue specimens possess the properties of CSCs. METHODS Twenty-five fresh liver tumor and adjacent nontumor tissue specimens resected from HCC patients were respectively digested into single liver cancer cells and single liver cells, partial of which were analyzed for the proportion of CD133 expression using flow cytometry and the other single liver cancer cells were primarily cultured. Using flow cytometry, the CD133＋ and CD133- cells were sorted from the obtained liver cancer cells and then were comparably characterized the properties of cancer stem cells by colony-forming assay, sphere formation assay and xenograft tumorigenicity assay. RESULTS Expression of CD133 in 25 HCC tissue specimens ranged from 3. 8% to 8.3 %, comparing with nontumor liver tissues ranging from 0.1% to 0.4 %, and the difference was statistically significant （t=17.12,P〈0. 001）. When compared with CD133- cells, CD133＋ cells not only possessed the preferential ability to form tumor colony in vitro （25.2±0.8） % vs （7.6±0.8） % （t=81.95 ,P〈0. 001）, but also formed tumor spheroids in vitro （20.3±0.6）% vs （12.5± 1.4）G （t = 68. 17, P〈 0. 001 ）, had a greater ability to form xenograft in nude mice. CONCLUSION CD133＋ ceils isolated from HCC tissue specimens possess characteristics of CSCs.

关 键 词：肝细胞性肝癌 CDl133 肿瘤干细胞 特性 

分 类 号：R735.7[医药卫生—肿瘤]