| 摘要: |
| 目的通过调节糖原合酶激酶-3β(GSK-3β)来观察其对肝癌细胞迁移能力的影响,研究GSK-3β在肝癌细胞迁移过程中发挥的作用。方法分别应用GSK-3特异性抑制剂LiCl和SB216763抑制GSK-3β的活性,或转染GSK-3β WT、GSK-3β S9A、GID5-6质粒影响GSK-3β的活性,通过划痕实验、小室迁移实验、微管组织中心体(MTOC)实验等观察处理前后肝癌细胞系SMMC-7721和Hep3B迁移能力的改变,并用免疫细胞化学法观察肝癌细胞迁移后磷酸化GSK-3β(pGSK-3β)的分布。结果划痕实验发现,用LiCl和SB216763处理后,SMMC-7721细胞的相对迁移率分别下降36.44%和41.78%,Hep3B细胞的相对迁移率都下降26.66%。转染GSK-3β WT后,GSK-3β和pGSK-3β表达上调,SMMC-7721细胞相对迁移率降为61.27%；转染GSK-3β S9A后,GSK-3β表达上调,pGSK-3β变化不明显,SMMC-7721细胞相对迁移率降为38.61%；转染GID5-6后,GSK-3β变化不明显,pGSK-3β表达增多,SMMC-7721细胞相对迁移率降为36.49%。免疫细胞化学可见,在正常培养的条件下肝癌细胞发生迁移3 h后,细胞内pGSK-3β呈现出在细胞前沿边缘附集的现象；而加入LiCl后,pGSK-3β的分布则未呈现这种边缘附集的现象。MTOC实验发现,未加入抑制剂时Hep3B细胞MTOC阳性率在60%以上,而应用GSK-3β抑制剂后MTOC阳性率明显下降。结论GSK-3β在肝癌细胞的迁移过程中起到重要作用,应用针对GSK-3β的特异性抑制药物能够影响肝癌细胞的极性化,降低肝癌细胞的迁移能力。 |
| 关键词: 糖原合酶激酶-3β 肝肿瘤 肝细胞癌 细胞迁移 |
| DOI：10.3724/SP.J.1008.2010.028 |
| 投稿时间：2008-11-11修订日期：2009-12-02 |
| 基金项目: |
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| Role of GSK-3β in hepatocellular carcinoma cell migration |
| WANG Jian-fei1,HOU Ying2,GE Rui-liang1,WANG Yi-zheng2,SHEN Feng1*,WU Meng-chao1 |
| (1.Eastern Hepatobiliary Surgery Hospital,Second Military Medical University,Shanghai 200438,China;2.Institute of Neuroscience,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,Shanghai 200031,China) |
| Abstract: |
| ObjectiveTo study the effects of GSK-3β on the migration ability of hepatocellular carcinoma cells.MethodsInhibition tests with specific inhibitors of GSK-3 (LiCl and SB216763) and transfection with different plasmids(GSK-3β WT,GSK-3β S9A,and GID5-6) were used to alter the activity of GSK-3β.The migration ability of SMMC-7221 and Hep3B cells was observed using wound healing assay,transwell assay and the microtubule-organizing center (MTOC) test before and after treatment.Immunocytochemistry was used to investigate the distribution of phospho-GSK-3β.ResultsWound healing assay showed that LiCl and SB216763 decreased the migration of SMMC-7721 cells by 36.44% and 41.78%,respectively; and decreased the migration of Hep3B cells by both 26.66%.Transfection with GSK-3β WT increased the expression of GSK-3β and pGSK-3β,and the migration rate of SMMC-7721 cells decreased to 61.27%. Transfection with GSK-3β S9A increased GSK-3β expression and had no noticeable influence on pGSK-3β expression,and the migration rate of SMMC-7721 cells decreased to 38.61%.Transfection with GID5-6 had no noticeable influence on GSK-3β and increased pGSK-3β expression,and the migration rate decreased to 36.49%.Immunocytochemical staining showed that LiCl blocked pGSK-3β accumulation on cell edge as that in normally cultured cells.MTOC test showed that the MTOC positive rate was above 60% in Hep3B cells before adding inhibitors of GSK-3β,and it decreased after exposure to the inhibitors.ConclusionGSK-3β plays an important role in migration of hepatocellular carcinoma cells.Inhibitors of GSK-3β can affect the polarization and decrease the migration ability of hepatocellular carcinoma cells. |
| Key words: GSK-3β liver neoplasms hepatocellular carcinoma cell migration |
