【摘要】 目的：溴结构域PHD指转录因子（bromodomain PHD finger transcription factor, BPTF）的异常表达与多种癌症的发生发展相关。本研究拟通过类器官模型探讨BPTF在胆管癌（cholangiocarcinoma, CCA）中的功能及其作为治疗靶点的潜力。方法：利用GEPIA数据库分析BPTF基因在CCA组织和正常组织中的表达差异，并通过免疫组化（immunohistochemistry, IHC）检测BPTF蛋白的表达水平。分离患者组织来源的CCA细胞，在基质胶三维环境中培养形成CCA类器官，并通过苏木精-伊红（hematoxylin-eosin, HE）和IHC染色对其进行组织学鉴定。用慢病毒介导的RNA干扰技术构建BPTF稳定敲减的CCA类器官模型，通过CCK-8法和EdU实验评估类器官的增殖能力，统计类器官形成的大小和数量以分析类器官的形成能力，通过药物敏感性实验探究BPTF对CCA化疗耐药中的影响。结果：GEPIA数据库分析和IHC检测结果表明，CCA组织中BPTF的mRNA和蛋白表达水平均显著升高（P<0.05或P<0.01）。成功构建了3例CCA类器官模型，HE和IHC染色结果证实其保留了原发组织的组织学特征。在CCA类器官中敲减BPTF显著抑制了类器官的增殖和形成（P<0.05或P<0.01）。此外，敲减BPTF增强了类器官对化疗药物吉西他滨和顺铂的敏感性。结论：下调BPTF的表达抑制了CCA类器官的增殖和形成能力，并增强了CCA类器官对化疗药物的敏感性。这提示BPTF可能在CCA进展中发挥重要作用，并有望成为CCA治疗的新靶点。更多还原

【Abstract】 AIM: Abnormal expression of bromodomain PHD finger transcription factor(BPTF) has been linked to the occurrence and progression of various cancers. This study aims to investigate the role of BPTF in cholangiocarcinoma(CCA) using patient-derived organoid(PDO) models and to evaluate its potential as a therapeutic target. METHODS: The GEPIA database was used to analyze the differential expression of BPTF between CCA and normal tissues. Immunohistochemistry(IHC) was performed to detect BPTF protein expression. The CCA cells were isolated from patient tumor tissues and cultured in a 3D Matrigel to establish CCA organoids. These organoids were then histologically verified by hematoxylin-eosin(HE) staining and IHC. A stable BPTF knockdown CCA organoid model was established using lentivirus-mediated RNA interference. Organoid proliferation was assessed using CCK-8 and EdU assays. Furthermore, the size and number of organoids were statistically analyzed to assess their formation ability. Drug sensitivity assays were conducted to assess the potential role of BPTF in chemoresistance of CCA. RESULTS: The BPTF expression at mRNA and protein levels was significantly increased in CCA tissues(P<0. 05 or P<0. 01). Three CCA organoid models were successfully established. Moreover, HE and IHC staining confirmed that these PDOs retained the histological characteristics of the original tissue. Knockdown of BPTF in CCA organoids significantly inhibited organoid proliferation and formation(P<0. 05 or P<0. 01). Furthermore, BPTF knockdown increased the sensitivity of CCA organoids to the chemotherapy drugs, such as gemcitabine and cisplatin. CONCLUSION: Down-regulation of BPTF expression inhibits the proliferation and formation ability of CCA organoids, and enhances their sensitivity to chemotherapy drugs. These results suggest that BPTF plays a significant role in the progression of CCA and holds promise as a potential therapeutic target for treatment of CCA.更多还原