The treatment of dopamine agonists (DA) resistant prolactinomas remains a formidable challenge, as the mechanism of resistance is still unclear, and there are currently no viable alternative drug therapies available. This study seeks to investigate the mechanism of DA resistance in prolactinomas and identify new potentially effective drugs. To explore the mechanism of DA resistance in prolactinomas, this study conducted transcriptome sequencing analysis on 27 cases of DA-resistant prolactinomas and 10 cases of sensitive prolactinomas. In addition, single-cell sequencing analysis was performed on 3 cases of DA-resistant prolactinomas and 3 cases of sensitive prolactinomas. Furthermore, to screen for potential therapeutic drugs, the study successfully established an organoids model for DA-resistant prolactinomas and screened 180 small molecule compounds using 8 organoids. The efficacy of the identified drugs was verified through various assays, including CCK-8, colony formation, CTG, and flow cytometry, and their mechanisms of action were confirmed through WB and IHC. The effectiveness of the identified drugs was evaluated both in vitro and in vivo. The results of transcriptome sequencing and single-cell sequencing analyses showed that DA resistance in prolactinomas is associated with the upregulation of the Focal Adhesion (FA) signaling pathway. Additionally, immunohistochemical validation revealed that FAK and Paxillin were significantly upregulated in DA-resistant prolactinomas. Screening of 180 small molecule compounds using 8 organoids identified Genistein as a potentially effective drug for DA-resistant prolactinomas. Experimental validation demonstrated that Genistein inhibited the proliferation of pituitary tumor cell lines and organoids and promoted apoptosis in pituitary tumor cells. Moreover, both the cell sequencing results and WB validation results of the drug-treated cells indicated that Genistein exerts its anti-tumor effect by inhibiting the FA pathway. In vivo, experiments also showed that Genistein can inhibit subcutaneous tumor formation. DA resistance in prolactinomas is associated with upregulation of the Focal Adhesion (FA) signaling pathway, and Genistein can exert its anti-tumor effect by inhibiting the expression of the FA pathway.

中文翻译：

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多巴胺激动剂耐药催乳素垂体神经内分泌肿瘤的耐药机制及新药探索

多巴胺激动剂（DA）耐药的催乳素瘤的治疗仍然是一个巨大的挑战，因为耐药机制仍不清楚，而且目前没有可行的替代药物疗法。本研究旨在探讨泌乳素瘤中 DA 耐药的机制并确定新的潜在有效药物。为了探讨泌乳素瘤中DA耐药的机制，本研究对27例DA耐药泌乳素瘤和10例敏感泌乳素瘤进行转录组测序分析。此外，还对3例DA耐药性泌乳素瘤和3例敏感性泌乳素瘤进行了单细胞测序分析。此外，为了筛选潜在的治疗药物，该研究成功建立了DA耐药泌乳素瘤的类器官模型，并利用8种类器官筛选了180个小分子化合物。通过CCK-8、集落形成、CTG和流式细胞术等多种检测验证了所鉴定药物的功效，并通过WB和IHC证实了其作用机制。在体外和体内评估了所鉴定药物的有效性。转录组测序和单细胞测序分析结果表明，泌乳素瘤中的 DA 耐药性与粘着斑 (FA) 信号通路的上调有关。此外，免疫组织化学验证显示，FAK 和 Paxillin 在 DA 抗性泌乳素瘤中显着上调。使用 8 种类器官筛选 180 种小分子化合物，确定金雀异黄素是治疗 DA 抗性催乳素瘤的潜在有效药物。实验验证表明，Genistein 可抑制垂体肿瘤细胞系和类器官的增殖，并促进垂体肿瘤细胞的凋亡。此外，药物处理细胞的细胞测序结果和WB验证结果均表明Genistein通过抑制FA途径发挥抗肿瘤作用。在体内，实验还表明金雀异黄素可以抑制皮下肿瘤的形成。泌乳素瘤中的DA抵抗与粘着斑（FA）信号通路的上调有关，金雀异黄素可以通过抑制FA通路的表达来发挥其抗肿瘤作用。