Sun Ding, Qin Lei, Xu Yang, Liu Jian-Xia, Tian Li-Ping, Qian Hai-Xin
Ding Sun, Lei Qin, Yang Xu, Jian-Xia Liu, Li-Ping Tian, Hai-Xin Qian, Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu Province, China.
World J Gastroenterol. 2014 Jun 14;20(22):6974-80. doi: 10.3748/wjg.v20.i22.6974.
To determine the influence of Adriamycin (ADM) on the changes in Nanog, Oct4, Sox2, as well as, in ARID1 and Wnt5b expression in liver cancer stem cells.
The MHCC97-L and HCCLM3 liver cancer cell lines were selected as the cell models in this study, and were routinely cultured. The 50% lethal dose (LD50) in the cell lines was detected by the MTT assay. Expression changes in liver cancer stem cell related genes (Nanog, Oct-4, Sox2, ARID1, and Wnt5b) were detected by western blot following treatment with ADM (LD50).
The LD50 of ADM in MHCC97-L cells was lower than that in HCCLM3 cells (0.4123 ± 0.0236 μmol/L vs 0.5259 ± 0.0125 μmol/L, P < 0.05). Wnt5b and Nanog were expressed in both MHCC97-L and HCCLM3 cells, while only Sox2 was expressed in HCCLM3 cells. However, neither ARID1A nor Oct4 was detected in these two cell lines. Genes, related to the stem cells, showed different expression in liver cancer cells with different metastatic potential following treatment with ADM (LD50). Wnt5b protein increased gradually within 4 h of ADM (LD50) treatment, while Nanog decreased (P < 0.05). After 12 h, Wnt5b decreased gradually, while Nanog increased steadily (P < 0.05). In addition, only Sox2 was expressed in HCCLM3 cells with high metastatic potential following ADM (LD50) treatment. The expression of Sox2 increased gradually with ADM (LD50) in HCCLM3 cells (P < 0.05).
ADM increased the death rate of MHCC97-L and HCCLM3 cells, while the growth suppressive effect of ADM was higher in MHCC97-L cells than in HCCLM3 cells.
确定阿霉素（ADM）对肝癌干细胞中Nanog、Oct4、Sox2以及ARID1和Wnt5b表达变化的影响。
本研究选择MHCC97-L和HCCLM3肝癌细胞系作为细胞模型，并进行常规培养。通过MTT法检测细胞系中的50%致死剂量（LD50）。在用ADM（LD50）处理后，通过蛋白质印迹法检测肝癌干细胞相关基因（Nanog、Oct-4、Sox2、ARID1和Wnt5b）的表达变化。
ADM在MHCC97-L细胞中的LD50低于HCCLM3细胞（0.4123±0.0236μmol/L对0.5259±0.0125μmol/L，P<0.05）。Wnt5b和Nanog在MHCC97-L和HCCLM3细胞中均有表达，而仅Sox2在HCCLM3细胞中表达。然而，在这两种细胞系中均未检测到ARID1A和Oct4。在用ADM（LD50）处理后，与干细胞相关的基因在具有不同转移潜能的肝癌细胞中表现出不同的表达。在ADM（LD50）处理4小时内，Wnt5b蛋白逐渐增加，而Nanog减少（P<0.05）。12小时后，Wnt5b逐渐减少，而Nanog稳步增加（P<0.05）。此外，在用ADM（LD50）处理后，仅Sox2在具有高转移潜能的HCCLM3细胞中表达。在HCCLM3细胞中，Sox2的表达随ADM（LD50）逐渐增加（P<0.05）。
ADM增加了MHCC97-L和HCCLM3细胞的死亡率，而ADM对MHCC97-L细胞的生长抑制作用高于HCCLM3细胞。
