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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。
PURPOSE More and more studies have demonstrated the presence of cancer stem cells (CSCs) in hepatoma cell lines. This study aimed to investigate whether CD133 + cells isolated from hepatocellular carcinoma (HCC) tissue samples have CSCs characteristics. Methods Twenty-five fresh HCC tissues and corresponding para-cancerous tissues obtained by surgery from Department of Hepatobiliary Surgery, Affiliated Tumor Hospital, Guangxi Medical University, 2014-02-01-2015-06-30 were digested into single hepatocarcinoma cells and single hepatocytes by enzyme digestion, The expression of CD133 in some single hepatocarcinoma cells and single hepatocytes was detected by flow cytometry. The remaining primary hepatoma cells were used for primary culture. The cultured hepatoma cells were sorted into CD133 + cells and CD133- cells by flow cytometry. The results of plate clone formation assay, tumor formation assay and xenograft tumor formation assay were compared Characteristics of CSCs in two groups of cells. Results The expression of CD133 in 25 HCC tissues was 3.8% -8.3% with a mean of 5.8 ± 1.6%, while the expression of CD133 in adjacent tissues was 0.1% -0.4% with a mean of (0.2 ± 0.1) %, The difference between the two was statistically significant, t = 17.12, P <0.001. The mean clonality rates of CD133 + and CD133- cells were (25.2 ± 0.8)% and (7.6 ± 0.8)% respectively, with significant difference between the two groups (t = 81.95, P <0.001). The average percentage of ball formation of CD133 + and CD133- cells was (20.3 ± 0.6)% and (12.5 ± 1.4)%, respectively, with significant difference between the two groups (t = 68.17, P <0.001). The ability of CD133 + cells to form xenografts in nude mice was significantly higher than that of CD133- cells. Conclusion The CD133 + cells isolated from HCC tissue samples have obvious characteristics of CSCs.