阿帕替尼通过抑制上皮-间充质转化阻遏肝癌细胞侵袭和转移的研究

Apatinib repressed the invasion and metastasis of hepatoceilular carcinoma cells by inhibiting epithelial-mesenchymal transformation

目的观察抗血管生成靶向药物阿帕替尼对肝癌细胞侵袭转移的影响,探讨其机制。方法将肝细胞癌细胞株SMMC-7721分为对照组和阿帕替尼处理组,观察细胞形态学变化;通过细胞计数试剂盒(CCK-8)试验观察阿帕替尼对SMMC-7721细胞增殖的影响;划痕实验观察阿帕替尼对SMMC-7721细胞侵袭能力的影响;Transwell试验观察阿帕替尼对SMMC-7721细胞转移能力的影响;实时定量聚合酶链反应(Real-timePCR)和Westernblot检测上皮-间充质转化(EMT)相关基因CDH1、CDH2、TJP1、VIM、E盒结合锌指蛋白1(ZEB1)、CLDN1、SNAI2、SNAI1、CTNNB1的表达变化。结果阿帕替尼处理后SMMC-7721细胞发生上皮表型的改变,增殖、侵袭和转移能力均降低。Real-timePCR和Westernblot结果显示经阿帕替尼处理后,SMMC-7721细胞中E-钙黏蛋白(E-cadherin)和ZO-1蛋白表达升高,N-cadherin、波形蛋白(Vimentin)、ZEB1、Slug表达降低,差异有统计学意义(CDH1,P=0.000;TJP1,P=0.002;CDH2,P=0.017;VIM,P=0.006;ZEB1,P=0.045;SNAI2,P=0.000)。结论阿帕替尼可抑制SMMC-7721细胞由上皮表型向间质表型转化(EMT),进而抑制肝癌细胞的侵袭和转移能力。

Objective To study the effect of anti-angiogenic targeting drug apatinib on invasion and metastasis of hepatoma cells and explore related mechanisms.Methods The hepatocellular carcinoma cell line SMMC-7721 was divided into control group and apatinib treatment group,and morphological changes of cells were observed.Cell counting kit-8 (CCK-8) assay was used to observe the proliferation effect of apatinib on SMMC-7721 cells,and the effects of apatinib on the invasion and metastasis of SMMC-7721 ceUs were observed by Transwell assay and wound healing assay respectively.Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blotting were used to detect the expression changes of epithelial-mesenchymal transition (EMT)-related genes CDH1,CDH2,TJP1,VIM,zinc finger E-box binding protein 1 (ZEB1),CLDN1,SNAI2,SNAI1 and CTNNB1.Results Under the effect of apatinib,the SMMC-7721 cells changed to be epithelial phenotypic,and the capabilities of proliferation,invasion and metastasis were all reduced.The results of RT-qPCR and Western blotting showed that the mRNA expression levels of CDH1 and TJP1 were increased,while the mRNA levels of CDH2,VIM,ZEB1 and Slug were decreased,and the differences were statistically significant (for CDH1,P =0.000;for TJP1,P =0.002;for CDH2,P =0.017;for VIM,P =0.006;for ZEB1,P =0.045;for SNAI2,P =0.000).Conclusion Apatinib can inhibit the transformation of hepatocellular carcinoma cell line SMMC-7721 from epithelial phenotype to mesenchymal phenotype and repress the invasion and metastasis of cancer cells.