Bufalin对食管癌ECA109细胞中FAK活化和上皮-间质转化的影响

Effects of Bufalin on the active state of FAK and epithelial-mesenchymal transition in esophageal squamous cell carcinoma ECA109 cells

目的探讨Bufalin对食管鳞状细胞癌ECA109细胞中FAK活化和上皮-间质转化(epithelial-mesenehymal transition,EMT)的影响。方法采用RT-PCR法检测不同浓度Bufalin对食管鳞状细胞癌ECA109细胞中FAK、E-cadherin、vimentin mRNA表达的影响。Western blot法检测不同浓度Bufalin对ECA109细胞中FAK活化水平及FAK、E-cadherin、vimentin蛋白表达的影响。采用Transwell小室实验检测不同浓度Bufalin对ECA109细胞迁移与侵袭能力的影响。结果 RT-PCR结果表明Bufalin抑制FAK、E-cadherin、vimentin的mRNA表达。Western blot结果表明Bufalin抑制E-cadherin、vimentin的表达和FAK的活化。Transwell实验结果表明Bufalin可以抑制ECA109细胞的迁移及侵袭。药物干预组(20、40、60、80、100 nmol/L Bufalin及PF562271组)与阳性对照组相比,Transwell迁移实验结果显示穿过基膜的细胞个数由107.00±8.19下降至78.67±3.06、61.67±3.06、42.67±3.512、24.33±2.517、10.33±3.215、9.00±2.65;Transwell侵袭实验结果显示穿过基膜的细胞个数由127.67±8.02下降至102.33±4.51、87.33±7.10、73.00±4.58、57.33±2.52、39.00±3.61、37.33±2.52,差异均有统计学意义(P<0.05)。结论 Bufalin可以抑制食管鳞状细胞癌中FAK的活化,提示Bufalin可能是通过下调FAK来抑制食管鳞状细胞癌EMT过程及食管癌的迁移及侵袭。

Purpose To investigate the effects of Bufalin on the active state of FAK and epithelial-mescnchymal transition （EMT） in ECA109 cells. Methods The mRNA expression of FAK, E-cadherin, vimentin in different concentrations of Bufalin in ECA109 cells was examined by RT-PCR. The protein expression of FAK, E-cadherin, vimentin and the active state of FAK in different concentra- tions of Bufalin in ECA109 cells were examined by Western blot. Transwell chamber experiment was used to detect migration and inva- sion ability in different concentrations of Bufalin in ECA109 cells. Results RT-PCR results showed that Bufalin decreased the mRNA expression of FAK, increased the mRNA expression of E-cadherin, and decreased the mRNA expression of vimentin. Western blot results showed that Bufalin decreased the active state of FAK, increased the protein expression of E-cadherin, and decreased the protein expression of vimentin. The results indicated that Bufalin inhibited cell migration and invasion in ECA109 cells. Compared with the positive control group, 20, 40, 60, 80, 100 nmol/L Bufalin and PF562271 group cell numbers of Transwell migration experiment crossed the basement membrane decreased from 107.00 ± 8. 19 to 78.67± 3.06, 61.67 ± 3.06, 42. 67± 3. 512, 24. 33 ± 2. 517, 10. 33 ±3. 215 and 9.00 ±2. 65, cell numbers of Transwell invasion experiment decreased from 127.67 ± 8.02 drop to 102. 33 ± 4.51, 87.33 ±7. 10, 73.00 ±4. 58, 57.33 ±2.52, 39. 00 ±3.61 and 37.33±2. 52, respectively, with statistically significant differences （P 〈 0. 05 ）. Conclusion Bufalin can inhibit the activation of FAK and EMT process in ECA109 cells, indicating that Bufalin may suppress the migration and invasion of ECA109 cells through down-regulating FAK.