Tessarollo Nayara Gusmão, Guimarães Isabella Dos Santos, Dos Santos Diandra Zipinotti, Henriques Taciane Barbosa, Lyra-Junior Paulo Cilas Morais, Carlos de Souza Josiany, Pimenta Tatiana Massariol, Martins Bárbara da Silva, Butzene Solenny Maria Silva, Padilha José Matheus Simões, Maciel Leide Laura Figueiredo, Almeida João Carlos de Aquino, Silva Ian Victor, Rangel Leticia Batista Azevedo
Biotechnology Program/RENORBIO, Health Sciences Center, Federal University of Espírito Santo, Vitória, Espírito Santo, Brazil.
Division of Clinical Research and Technological Development, Brazilian National Cancer Institute (INCA), Rio de Janeiro, Brazil.
Front Pharmacol. 2025 Jun 4;16:1566330. doi: 10.3389/fphar.2025.1566330. eCollection 2025.
Chemoresistance and disease relapses in epithelial ovarian cancer (EOC) highlight the need for novel therapeutic strategies. Here, we investigated phosphodiesterase 7A (PDE7A) as a potential target in ovarian cancer treatment.
Gene expression was performed by RNA sequencing data comparing high-grade serous ovarian carcinoma (HGSOC) and fallopian tube samples. The PDE7 inhibitor BRL 50481, alone or combined with paclitaxel (PTX), was tested in drug-sensitive A2780 and multi-resistant OVCAR3 cells by Diphenyltetrazolium bromide (MTT) assay. To validate data from the high throughput RNA-sequencing assays, RT-qPCR and Immunoblotting were performed. Cytokine expression was analyzed by RT-qPCR and the quantification was obtained by ELISA. Scanning and Transmission Electron Microscopy were also carried out.
MTT assays revealed that while BRL 50481 reduced metabolic cellular viability (MCV) in A2780 (IC50 = 200 μM), its combination with PTX decreased MCV in both lines, reducing PTX IC50 by 103- and 625-fold in A2780 and OVCAR3, respectively. PDE7 inhibition suppressed the PI3K/AKT/mTOR pathway, upregulated the pro-apoptotic protein Bcl-2 Associated X-protein (BAX) in A2780, and increased IL-6 expression in OVCAR3. Pretreatment with BRL 50481 followed by PTX downregulated vimentin and octamer-binding transcription factor (OCT4), while inducing morphological changes and mitochondrial cristae alterations. Inhibiting PDE7 can enhance the paclitaxel-induced apoptosis by promoting mitochondrial dysfunction and suppressing survival pathways, thereby improving ovarian cancer treatment efficacy. The results need to be validated in additional in vivo models.
上皮性卵巢癌（EOC）中的化疗耐药和疾病复发凸显了新型治疗策略的必要性。在此，我们研究了磷酸二酯酶7A（PDE7A）作为卵巢癌治疗潜在靶点的可能性。
通过对高级别浆液性卵巢癌（HGSOC）和输卵管样本的RNA测序数据进行基因表达分析。使用二苯基四氮唑溴盐（MTT）法，在药物敏感的A2780细胞和多药耐药的OVCAR3细胞中测试PDE7抑制剂BRL 50481单独使用或与紫杉醇（PTX）联合使用的效果。为了验证高通量RNA测序分析的数据，进行了逆转录定量聚合酶链反应（RT-qPCR）和免疫印迹分析。通过RT-qPCR分析细胞因子表达，并通过酶联免疫吸附测定（ELISA）进行定量。还进行了扫描电子显微镜和透射电子显微镜检查。
MTT分析显示，虽然BRL 50481降低了A2780细胞的代谢细胞活力（MCV）（半数抑制浓度[IC50]=200μM），但其与PTX联合使用降低了两株细胞系的MCV，在A2780细胞和OVCAR3细胞中分别将PTX的IC50降低了103倍和625倍。PDE7抑制作用抑制了磷脂酰肌醇-3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白（PI3K/AKT/mTOR）信号通路，上调了A2780细胞中促凋亡蛋白Bcl-2相关X蛋白（BAX）的表达，并增加了OVCAR3细胞中白细胞介素-6（IL-6）的表达。先用BRL 50481预处理再用PTX处理可下调波形蛋白和八聚体结合转录因子（OCT4）的表达，同时诱导形态学变化和线粒体嵴改变。抑制PDE7可通过促进线粒体功能障碍和抑制生存信号通路增强紫杉醇诱导的细胞凋亡，从而提高卵巢癌治疗效果。这些结果需要在更多体内模型中进行验证。
