TNF-α对AKT-TWIST1信号通路诱导乳头状甲状腺癌细胞株上皮间质转化的影响

TNF-α induced EMT through AKT-TWIST1 pathway in PTC cell line

目的观察TNF-α是否可诱导乳头状甲状腺癌细胞株BCPAP的上皮间质转化,并探讨AKT-TWIST1信号通路在其中的作用。方法采用Western检测TNF-α刺激后BCPAP细胞中EMT标志物E-cadherin,N-cadherin,Vimentin以及AKT,TWIST1的蛋白水平变化。采用RT-PCR检测TWIST1的mRNA水平变化。采用划痕愈合实验和Transwell侵袭实验检测AKT抑制后TNF-α对BCPAP侵袭转移能力的影响。结果 TNF-α作用下BCPAP的E-cadherin的蛋白表达下降,而N-cadherin和Vimentin的蛋白水平上升(P<0.05),同样的,TWIST1的mRNA水平上升(P<0.05),AKT及TWIST1的蛋白水平上升(P<0.05);应用AKT抑制剂后TWIST1的mRNA及蛋白表达降低,且BCPAP细胞的侵袭转移能力下降。结论 TNF-α可能通过ATK-TWIST1信号通路诱导EMT,进而促进人乳头状甲状腺癌细胞株BCPAP的侵袭转移。

Objective To investigate the effects of TNF-α on the EMT in PTC cell line BCPAP, confirmed wether the AKT-TWIST1 pathway was involved in these progressions. Methods The expression levels of proteins of EMT makers E-cadherin, N-cadherin and Vimentin, AKT, TWSIT1 were analyzed using Western blot. The expression level of mRNA of TWIST1 was analyzed using RT-PCR. The effect of TNF-α on cell migration and invasion after AKT inhibiting by HY-89 was assessed by Wound-healing and Transwell assay. Results After the involvement of TNF-α, the protein expression of E-cadherin of BCPAP decreased, while the protein expressions of N-cadherin and Vimentin increased(P<0.05). The mRNA of TWIST1 increased(P<0.05), and the protein expressions of AKT and TWIST1 increased(P<0.05). After AKT inhibitors were applied, TWIST1 protein expression was decreased and cell invasive and metastasis capacity was decreased. Conclusion Our results showed that TNF-α could induce EMT to promote the invasive and metastasis capacity of human papillary thyroid carcinoma cell line BCPAP through the ATK-TWIST1 signaling pathway.