POU5F1B is responsible for the acquired resistance to dabrafenib in papillary thyroid cancer cells with the BRAF V600E mutation.

IF 3 3区医学 Q2 ENDOCRINOLOGY & METABOLISM

Endocrine Pub Date : 2025-01-01 Epub Date: 2024-08-13 DOI:10.1007/s12020-024-03994-y

Jun Li, Yafeng Yu

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Abstract

Background: Dabrafenib, an inhibitor of the B-Raf proto-oncogene (BRAF) V600E mutant, has become the major drug for targeted therapy of papillary thyroid cancer (PTC) with the BRAF V600E mutant; however, acquired resistance is inevitable.

Objective: To identify key transcription factors (TFs) involved in dabrafenib resistance and identify targets to reverse dabrafenib resistance.

Methods: Dabrafenib-resistant PTC cell lines BCPAP/DabR and K1/DabR were established, and phenotypic assays were performed to validate the malignant phenotype. RNA sequencing and bioinformatics analyses were used to identify differentially expressed genes (DEGs) and screen TFs involved in resistant phenotype-related pathways. The role of the key TF POU5F1B in dabrafenib resistance was further validated using gene gain-and-loss assays.

Results: BCPAP/DabR and K1/DabR were resistant to dabrafenib, with a resistance index of 5-8. Resistant cells exhibited slower proliferation, strong migration, and spheroid-forming abilities. RNA sequencing screened 6233 DEGs in the resistant group, including 2687 protein-coding RNA (mRNA). Venn analysis indicated that three genes, E2F2, WNT4, and POU5F1B, were involved in resistant phenotype-related pathways and were included in the TF regulatory network. Four TFs of the three genes, POU5F1B, TBX4, FOXO4, and FOXP3, were validated, and POU5F1B showed the highest validated fold-change. Overexpression of POU5F1B in sensitive cells resulted in resistance to dabrafenib and induced a malignant phenotype, whereas silencing it sensitized the resistant cells and reversed the resistant phenotype.

Conclusion: This study successfully established two dabrafenib-resistant PTC cell lines, and POU5F1B could be a potential target for reversing dabrafenib resistance.

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POU5F1B 是 BRAF V600E 突变的甲状腺乳头状癌细胞对达拉菲尼产生获得性耐药性的原因。

背景：达拉菲尼是B-Raf原癌基因（BRAF）V600E突变体的抑制剂，已成为BRAF V600E突变体甲状腺乳头状癌（PTC）靶向治疗的主要药物；然而，获得性耐药性是不可避免的：鉴定参与达拉菲尼耐药的关键转录因子（TFs），并确定逆转达拉菲尼耐药的靶点：方法：建立达拉非尼耐药的PTC细胞系BCPAP/DabR和K1/DabR，并进行表型检测以验证恶性表型。利用RNA测序和生物信息学分析鉴定差异表达基因（DEGs）并筛选参与耐药表型相关通路的TFs。利用基因增益和缺失试验进一步验证了关键TF POU5F1B在达拉菲尼耐药中的作用：结果：BCPAP/DabR和K1/DabR对达拉菲尼耐药，耐药指数为5-8。耐药细胞的增殖速度慢、迁移能力强、球形细胞形成能力强。RNA 测序筛选了耐药组中的 6233 个 DEGs，包括 2687 个蛋白编码 RNA（mRNA）。维恩分析表明，E2F2、WNT4和POU5F1B三个基因参与了抗性表型相关通路，并被纳入TF调控网络。这三个基因中的四个TFs，即POU5F1B、TBX4、FOXO4和FOXP3得到了验证，其中POU5F1B的验证折合变化最高。在敏感细胞中过表达POU5F1B会导致对达拉菲尼耐药并诱导恶性表型，而沉默POU5F1B则会使耐药细胞敏感并逆转耐药表型：本研究成功建立了两种达拉非尼耐药的PTC细胞系，POU5F1B可能是逆转达拉非尼耐药的潜在靶点。

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来源期刊

Endocrine ENDOCRINOLOGY & METABOLISM-

CiteScore

6.50

自引率

5.40%

发文量

295

审稿时长

1.5 months

期刊介绍： Well-established as a major journal in today’s rapidly advancing experimental and clinical research areas, Endocrine publishes original articles devoted to basic (including molecular, cellular and physiological studies), translational and clinical research in all the different fields of endocrinology and metabolism. Articles will be accepted based on peer-reviews, priority, and editorial decision. Invited reviews, mini-reviews and viewpoints on relevant pathophysiological and clinical topics, as well as Editorials on articles appearing in the Journal, are published. Unsolicited Editorials will be evaluated by the editorial team. Outcomes of scientific meetings, as well as guidelines and position statements, may be submitted. The Journal also considers special feature articles in the field of endocrine genetics and epigenetics, as well as articles devoted to novel methods and techniques in endocrinology. Endocrine covers controversial, clinical endocrine issues. Meta-analyses on endocrine and metabolic topics are also accepted. Descriptions of single clinical cases and/or small patients studies are not published unless of exceptional interest. However, reports of novel imaging studies and endocrine side effects in single patients may be considered. Research letters and letters to the editor related or unrelated to recently published articles can be submitted. Endocrine covers leading topics in endocrinology such as neuroendocrinology, pituitary and hypothalamic peptides, thyroid physiological and clinical aspects, bone and mineral metabolism and osteoporosis, obesity, lipid and energy metabolism and food intake control, insulin, Type 1 and Type 2 diabetes, hormones of male and female reproduction, adrenal diseases pediatric and geriatric endocrinology, endocrine hypertension and endocrine oncology.