索拉菲尼对转化生长因子-β_1诱导的人肝癌细胞系SMCC-7721的上皮-间质转化的抑制作用

Effect of sorafenib on the epithelial-mesenchymal transition induced by transforming growth factor-β_1 in hepatocellular carcinoma cell line SMCC-7721

目的研究索拉菲尼对转化生长因子-β_1(TGF-β_1)诱导的人肝癌细胞系SMCC-7721上皮-间质转化(EMT)的抑制作用。方法将肝癌细胞系SMCC-7721分4组:空白组、模型组和低、高2个剂量实验组。空白组加入磷酸盐缓冲液(PBS)处理肝癌细胞,模型组用TGF-β_15 ng·mL-1处理肝癌细胞。在造模基础上,实验组分别加入索拉菲尼10,2μmol·mL-1处理肝癌细胞。各组细胞处理24 h后,划痕实验和Transwell侵袭实验分别观察不同处理组肿瘤细胞迁移和侵袭情况;以蛋白印记实验分别检测E-cadherin和Vimentin和蛋白表达情况。结果模型组、空白组与高剂量实验组的伤口愈合率分别为(53.72±2.83)%,(40.35±2.55)%,(30.38±3.00)%;这3组的穿过小室肿瘤细胞数量分别为130.25±14.39,84.50±8.81,37.75±6.75,模型组明显高于空白组,差异均有统计学意义(均P<0.05);高剂量实验组与模型组比较,这2项指标均明显降低,差异均有统计学意义(均P<0.05)。模型组与高剂量实验组的E-cadherin蛋白表达分别为0.80±0.18,1.56±0.24;这2组的Vimentin蛋白的表达分别为1.25±0.14,0.55±0.29,高剂量实验组与模型组相比,两者差异均有统计学意义(均P<0.05)。结论索拉菲尼可以抑制TGF-β_1诱导的肝癌细胞SMCC-7721的EMT和降低其迁移能力和侵袭性。

Objective To investigate the effect of sorafenib on transforming growth factor-β1（ TGF-β1）-mediated epithelial-mesenchymal transition（ EMT） and migration and invasion in hepatocellular carcinoma cell line SMCC-7721. Methods SMCC-7721 cells were divided into four groups： blank group,model group,and low,high doses of experimental groups. The liver cancer cells in the blank group were only treated with phosphate buffered saline（ PBS）,the cells in the model group were only treated with TGF-β15 ng·mL-1,and the tumor cells in the two doses of experimental groups were treated with sorafenib 10,2μmol·mL-1 in addition to culturing with TGF-β15 ng · mL-1. After culturing for 24 h,the ability of invasion and migration of SMCC-7721 cells was assessed by Wound healing assay and Transwell chamber assay.The levels of protein expression of E-cadherin and Vimentin were measured by Western blotting. Results The rate of wound healing in modelgroup,blank group and high dose experimental group were（ 53. 72 ± 2. 83） %,（ 40. 35 ± 2. 55） %,（ 30. 38 ± 3. 00） %;the number of invasive cells in the 3 groups were 130. 25 ± 14. 39,84. 50 ± 8. 81,37. 75 ± 6. 75,model group were significantly higher than that in the blank group（ all P〈0. 05）; the high dose experimental group were significantly lower than that in the model group（ all P〈0. 05）. The relative levels of protein expression of E-cadherin in model group and high dose experimental group were 0. 80 ± 0. 18,1. 56 ± 0. 24; the relative levels of protein expression of Vimentin in the 2 groups were 1. 25 ± 0. 14,0. 55 ± 0. 29,high dose experimental group were significantly different from that in the model group（ all P〈0. 05）. Conclusion Sorafenib can inhibit TGF-β1-mediated epithelial-EMT and migration and invasion in hepatocellular carcinoma cell line SMCC-7721.