Kim Young Hee, Kang Jin Seok
Department of Biomedical Laboratory Science, Namseoul University, Cheonan, Republic of Korea E-mail :
Asian Pac J Cancer Prev. 2013;14(11):6341-5. doi: 10.7314/apjcp.2013.14.11.6341.
To clarify the role of stem cells in hepatocarcinogenesis, glypican-3 (GPC-3) and E-cadherin expression was investigated in embryonic cell lineages. Mouse embryonic stem cells (ESCs), hepatic progenitor cells (HPCs) and hepatocyte like cells (HCs), representing 0, 22 and 40 days of differentiation, respectively, were treated in vitro with diethylnitrosamine (DEN) at four doses (0, 1, 5 and 15 mM; G1, G2, G3 and G4, respectively) for 24 h and GPC- 3 and E-cadherin expression was examined by relative quantitative real-time PCR and immunocytochemistry. GPC-3 mRNA expression was significantly different for G4 at day 0 (p<0.001) and for G4 at day 22 (p<0.01) compared with the control (G1). E-cadherin mRNA expression was significantly different for G3 and G4 at day 0 (p<0.05 and p<0.001, respectively), for G2 and G4 (p<0.05 and p<0.001, respectively) at day 22 and for G2 and G4 (p<0.01 and p<0.001, respectively) at day 40 compared with G1. Immunofluorescence staining for GPC-3 showed a membranous and/or granular expression in cytoplasm of ESCs and HPCs and granular and/or diffuse expression in cytoplasm of HCs, which were also stained by E-cadherin. DEN treatment increased GPC- 3 expression in ESCs, HPCs and HCs, with increase of E-cadherin expression. Taken together, the expression of GPC-3 was altered by DEN treatment. However, its expression pattern was different at the stage of embryo stem cells and its derived hepatic lineage cells. This suggests that GPC-3 expression may be modulated in the progeny of stem cells during their differentiation toward hepatocytes, associated with E-cadherin expression.
为阐明干细胞在肝癌发生中的作用，对胚胎细胞谱系中的磷脂酰肌醇蛋白聚糖-3（GPC-3）和E-钙黏蛋白表达进行了研究。分别代表分化0、22和40天的小鼠胚胎干细胞（ESC）、肝祖细胞（HPC）和肝细胞样细胞（HC），在体外分别用四种剂量（0、1、5和15 mM；分别为G1、G2、G3和G4）的二乙基亚硝胺（DEN）处理24小时，然后通过相对定量实时PCR和免疫细胞化学检测GPC-3和E-钙黏蛋白的表达。与对照组（G1）相比，G4在第0天（p<0.001）和G4在第22天（p<0.01）时GPC-3 mRNA表达有显著差异。与G1相比，G3和G4在第0天（分别为p<0.05和p<0.001）、G2和G4在第22天（分别为p<0.05和p<0.001）以及G2和G4在第40天（分别为p<0.01和p<0.001）时E-钙黏蛋白mRNA表达有显著差异。GPC-3的免疫荧光染色显示在ESC和HPC的细胞质中有膜性和/或颗粒状表达，在HC的细胞质中有颗粒状和/或弥漫性表达，E-钙黏蛋白也呈阳性染色。DEN处理增加了ESC、HPC和HC中GPC-3的表达，同时E-钙黏蛋白表达也增加。综上所述，DEN处理改变了GPC-3的表达。然而，其在胚胎干细胞及其衍生肝系细胞阶段的表达模式不同。这表明在干细胞向肝细胞分化过程中，其后代中GPC-3的表达可能受到调节，且与E-钙黏蛋白表达相关。
