通过抑制细胞周期蛋白依赖性激酶 1 提高胰腺癌的放射敏感性

IF 4.9 1区 医学 Q1 ONCOLOGY Radiotherapy and Oncology Pub Date : 2024-09-11 DOI:10.1016/j.radonc.2024.110531
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引用次数: 0

摘要

背景和目的克服放射抗性是胰腺导管腺癌(PDAC)面临的一项严峻挑战。我们的研究探讨了通过基因和药物抑制来靶向细胞周期蛋白依赖性激酶-1(CDK1),从而使 PDAC 细胞具有放射敏感性。材料与方法采用质谱和磷酸化蛋白质组学分析了与亲代对照相比具有放射抗性的 PDAC 细胞系(MIA PaCa-2 和 PANC-1)。我们查询了 TCGA PDAC 数据库的临床结果,并根据 CDK1 mRNA 表达的中位数对患者进行了二分。我们生成了一种基于微RNA的TET-on诱导型shRNA,以抑制两种PDAC细胞系中CDK1的表达。我们使用 PDAC 正位模型来测试 PDAC 肿瘤在接受或不接受强力霉素治疗时的辐射敏感性。我们使用选择性 CDK1 抑制剂 RO-3306 靶向激活 CDK1,然后使用免疫印迹、免疫细胞化学和克隆生成试验进行体外实验。结果磷蛋白组学分析表明,耐药克隆中磷酸化 CDK1(Tyr15)显著升高。我们发现,CDK1的高表达与PDAC患者较差的OS有关。辐射会增加 CDK1 磷酸化。在 MIA PaCa-2 和 PANC-1 细胞中,CDK1 抑制与放射治疗协同延缓体内肿瘤生长。通过 RO-3306 抑制 CDK1RO-3306 可使细胞明显进入 G2/M 期,并破坏辐射照射后的 DNA 修复。在体外,RO-3306 的预处理增强了 PDAC 细胞的放射敏感性。放疗靶向 CDK1 有望在 PDAC 治疗中得到进一步研究。
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Enhanced radiosensitivity of pancreatic cancer achieved through inhibition of Cyclin-dependent kinase 1

Background and purpose

Overcoming radioresistance is a critical challenge in pancreatic ductal adenocarcinoma (PDAC). Our study investigates the targeting of Cyclin-dependent kinase-1 (CDK1) through genetic and pharmaceutical inhibition to radiosensitize PDAC cells.

Materials and Methods

Mass spectrometry and phosphoproteomics were used to analyze engineered radiation-resistant PDAC cell lines (MIA PaCa-2 and PANC-1) compared to parental controls. The TCGA PDAC database was queried for clinical outcomes and patients were dichotomized based on the median CDK1 mRNA expression. We generated a microRNA-based TET-on inducible shRNA to inhibit CDK1 expression in two PDAC cell lines. We used an orthotopic model of PDAC to test the radiation sensitivity of PDAC tumors with or without doxycycline treatment. We targeted CDK1 activation with a selective CDK1 inhibitor, RO-3306, followed by in vitro experiments employing immunoblotting, immunocytochemistry, and clonogenic assays.

Results

Phosphoproteomics analysis revealed that phospho-CDK1 (Tyr15) was significantly elevated in the resistant clones. We found that high CDK1 expression was associated with worse OS in PDAC patients. Radiation exposure increased CDK1 phosphorylation. In MIA PaCa-2 and PANC-1 cells, CDK1 inhibition synergized with radiation therapy to delay tumor growth in vivo. CDK1 inhibition via. RO-3306 resulted in a significant shift of cells into the G2/M phase and disrupted DNA repair after radiation exposure. In vitro, pre-treatment with RO-3306 led to enhanced radiosensitivity of PDAC cells.

Conclusion

CDK1 plays a crucial role in PDAC radioresistance. Targeting CDK1 with radiotherapy holds promise for further investigation in PDAC treatment.

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来源期刊
Radiotherapy and Oncology
Radiotherapy and Oncology 医学-核医学
CiteScore
10.30
自引率
10.50%
发文量
2445
审稿时长
45 days
期刊介绍: Radiotherapy and Oncology publishes papers describing original research as well as review articles. It covers areas of interest relating to radiation oncology. This includes: clinical radiotherapy, combined modality treatment, translational studies, epidemiological outcomes, imaging, dosimetry, and radiation therapy planning, experimental work in radiobiology, chemobiology, hyperthermia and tumour biology, as well as data science in radiation oncology and physics aspects relevant to oncology.Papers on more general aspects of interest to the radiation oncologist including chemotherapy, surgery and immunology are also published.
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