PF-5274857阻断香烟烟雾诱导的上皮-间质转化的作用

目的 探讨Smoothened（Smo）抑制剂PF-5274857在香烟烟雾(CS)诱导所致的上皮-间质转化（EMT）转变中的作用, 为口腔鳞状细胞癌的临床治疗和预防提供依据。方法 以支气管上皮细胞株Beas-2b建立体外CS诱导EMT模型,实验分为预防实验和治疗实验两部分进行。预防实验:用3 μmol/L PF-5274857预刺激Beas-2b细胞2 h后用CS溶液培养8 d；治疗实验:CS培养8 d后用3 μmol/L PF-52748573处理Beas-2b细胞4 d。通过Western blot、免疫荧光检测细胞蛋白中EMT标志物E-钙黏蛋白（E-cadherin）、波形蛋白（vimentin）、α平滑肌肌动蛋白（α-SMA）的含量及位置变化，小室迁移实验测定治疗实验中细胞迁移能力改变。结果 PF-5274857预刺激2 h，间质标志物vimentin和α-SMA蛋白表达降低，上皮标志物E-cadherin表达升高。而已被CS诱导产生EMT改变的Beas-2b细胞经PF-5274857治疗4 d后部分恢复E-cadherin表达，并且vimentin和α-SMA蛋白表达降低，其升高的迁移能力也降低。 结论 PF-5274857可以预防和治疗由CS引起的Beas-2b细胞EMT。

STInhibition of Cigarettes Smoke-induced Epithelial to Mesenchymal Transition by the SMO Inhibitor PF-5274857 in Beas-2b Epithelial Cells

Objective To explore the effects of the Smoothened (Smo) inhibitor PF-5274857 on cigarette smoke (CS)-induced epithelial-mesenchymal transition (EMT) and apply a new idea fororal squamous cell carcinoma (OSCC) treatment. Methods Beas-2b cells were used to investigate the CS-induced EMT. Both pretreat and post-treat were performed in the study. In pretreat group, after being pretreated with 3 μmol/L PF-5274857 for 2 h, cells were cultured with CS for 8 d; In post-treat group, cells were treated by 3 μmol/L PF-5274857 for 4 d after CS cultrue. Western blot and immunofluorescence were applied to examine the EMT markers E-cadherin, vimentin, and smooth muscle actin α (α-SMA) in Beas-2b epithelial cells. The transwell culture system was used in migration assays. Results Pretreat Beas-2b cells with PF-5274857 for 2 h can prevent the CS-induced EMT for epithelial and mesenchymal markers, as well as migration capacity. Up regulated E-cadherin and down regulated vimentin and α-SMA were observed by Western blot. Furthermore Beas-2b cells induced by CS that underwent EMT showed increased E-cadherin and decreased vimentin and α-SMA after treatment with PF-5274857 for 4 d. Importantly, the elevated migration capacity level was also decreased. Conclusion The Smo inhibitor PF-5274857 has both preventive effect and therapeutic potential against CS-induced EMT.