食管腺癌（EAC）患者的临床结果糟糕，总体5年生存率为20%。吸烟是EAC的一个重要危险因素。WEE1是一种核激酶，在正常条件下负调节细胞周期，但其在EAC肿瘤发生和药物抗性中的作用尚不完全理解。免疫组化染色显示人EAC组织中WEE1的过度表达。经过Western blot和免疫荧光染色检测，烟碱、烟碱衍生亚硝酮或2%香烟烟雾提取物处理诱导EAC中WEE1蛋白的表达。qRT-PCR和报告基因实验表明，吸烟通过miR-195-5p下调在EAC中诱导WEE1表达。ATP-Glo细胞活力和克隆形成实验证实，WEE1抑制增加了EAC细胞对多西他赛的敏感性。使用TE-10吸烟机和EAC患者来源的异种移植小鼠模型，在体内证明吸烟诱导了WEE1蛋白的表达和对多西他赛的耐药性。MK-1775和多西他赛联合治疗提高了EAC患者来源的异种移植小鼠的生存率。我们的发现首次证明了吸烟诱导的WEE1过度表达通过miRNA失调在EAC的药物抗性中起到了至关重要的作用。WEE1抑制是克服药物抗性和靶向治疗难治性肿瘤细胞的一种有希望的治疗方法。© 2023作者。

Esophageal adenocarcinoma (EAC) patients have poor clinical outcomes, with an overall 5-year survival rate of 20%. Smoking is a significant risk factor for EAC. The role of WEE1, a nuclear kinase that negatively regulates the cell cycle in normal conditions, in EAC tumorigenesis and drug resistance is not fully understood. Immunohistochemistry staining shows significant WEE1 overexpression in human EAC tissues. Nicotine, nicotine-derived nitrosamine ketone, or 2% cigarette smoke extract treatment induces WEE1 protein expression in EAC, detected by western blot and immunofluorescence staining. qRT-PCR and reporter assay indicates that smoking induces WEE1 expression through miR-195-5p downregulation in EAC. ATP-Glo cell viability and clonogenic assay confirmed that WEE1 inhibition sensitizes EAC cells to docetaxel treatment in vitro. A TE-10 smoking machine with EAC patient-derived xenograft mouse model demonstrated that smoking induces WEE1 protein expression and resistance to docetaxel in vivo. MK-1775 and docetaxel combined treatment improves EAC patient-derived xenograft mouse survival in vivo. Our findings demonstrate, for the first time, that smoking-induced WEE1 overexpression through miRNA dysregulation in EAC plays an essential role in EAC drug resistance. WEE1 inhibition is a promising therapeutic method to overcome drug resistance and target treatment refractory cancer cells.© 2023 The Author(s).