斑蝥素调控IL-2Rα逆转皮肤T细胞淋巴瘤伏立诺他耐药作用研究

皮肤T细胞淋巴瘤（CTCL）是异质性T细胞侵袭皮肤为表现的恶性疾病，伏立诺他耐药是其治疗失败的重要原因。前期研究发现IL-2Rα在伏立诺他耐药CTCL中表达增高，敲低IL-2Rα可抵抗耐药性，表明IL-2Rα是抵抗CTCL对伏立诺他耐药的潜在靶点；经筛选发现斑蝥素可作用于IL-2Rα并抑制其蛋白表达，与伏立诺他联用能协同抑制耐药细胞生长。本研究将以伏立诺他耐药细胞为实验对象，通过IL-2Rα高、低表达细胞模型、尾静脉注射和异种移植瘤动物模型，深入研究斑蝥素抵抗伏立诺他耐药性与IL-2Rα的关系；运用IL-2Rα/HEK293细胞模型、亲和色谱法、分子动力学模拟和CRISPR/Cas9等技术，揭示斑蝥素与IL-2Rα的亲和作用及抵抗耐药性的核心作用位点；采用蛋白免疫印迹等技术在分子水平上研究斑蝥素对IL-2R信号通路的调控作用。本研究将为揭示斑蝥素逆转伏立诺他耐药的作用和机理提供实验依据。

Cutaneous T-cell lymphoma (CTCL) is characterized by the presence of malignant T cells in chronically inflamed skin lesions. Drug resistance to vorinotat (SAHA) makes it difficult for CTCL treatment. Our previous study has found that although IL-2Rα was over-expressed in both SAHA resistant cells and xenograft tumors, knock-down of IL-2Rα could reverse SAHA resistance. This demonstrates that IL-2Rα is a potential target for the reversal of SAHA resistance in CTCL. We has also found that cantharidin (CTD) could target IL-2Rα, inhibit its protein expression, and reverse the drug resistance of SAHA. In this study, SAHA-resistant cells were used as experimental objects, and the anti-tumor effects of CTD on SAHA resistant cells will be evaluated by cell models and animal models with over-expressed IL-2Rα and knockdown of IL-2Rα. Further experiments will be performed to investigate the affinity of CTD and IL-2Rα as well as its core action sites contributing to reversing SAHA resistance using IL-2Rα/HEK293 cells, affinity chromatography, molecular dynamics simulation and CRISPR/Cas9. Finally, western blot and other techniques will be used to study the regulation of CTD on IL-2Rα signaling pathway after acting on the core sites of IL-2Rα at the molecular level. This work may bring insights to the mechanism of SAHA resistance and provide an experimental basis for revealing the mechanism of CTD in reversing SAHA resistance of CTCL.