Jing Qian, Yanxia Ma, William M Tahaney, Cassandra L Moyer, Amanda Lanier, Jamal Hill, Darian Coleman, Negar Koupaei, Susan G Hilsenbeck, Michelle I Savage, Brent D G Page, Abhijit Mazumdar, Powel H Brown
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In this study, we investigated the potential of a novel phosphatase, NUDT5, as a potential therapeutic target for TNBC.</p><p><strong>Methods: </strong>The mRNA expression levels of NUDT5 in breast cancers were investigated using TCGA and METABRIC (Curtis) datasets. NUDT5 ablation was achieved through siRNA targeting and NUDT5 inhibition with the small molecule inhibitor TH5427. Xenograft TNBC animal models were employed to assess the effect of NUDT5 inhibition on in vivo tumor growth. Proliferation, death, and DNA replication assays were conducted to investigate the cellular biological effects of NUDT5 loss or inhibition. The accumulation of 8-oxo-guanine (8-oxoG) and the induction of γH<sub>2</sub>AX after NUDT5 loss was determined by immunofluorescence staining. The impact of NUDT5 loss on replication fork was assessed by measuring DNA fiber length.</p><p><strong>Results: </strong>In this study, we demonstrated the significant role of an overexpressed phosphatase, NUDT5, in regulating oxidative DNA damage in TNBCs. Our findings indicate that loss of NUDT5 results in suppressed growth of TNBC both in vitro and in vivo. This growth inhibition is not attributed to cell death, but rather to the suppression of proliferation. The loss or inhibition of NUDT5 led to an increase in the oxidative DNA lesion 8-oxoG, and triggered the DNA damage response in the nucleus. The interference with DNA replication ultimately inhibited proliferation.</p><p><strong>Conclusions: </strong>NUDT5 plays a crucial role in preventing oxidative DNA damage in TNBC cells. The loss or inhibition of NUDT5 significantly suppresses the growth of TNBCs. 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Treatment options for women with TNBC tumors are limited, unlike those with ER-positive tumors that can be treated with hormone therapy, or those with HER2-positive tumors that can be treated with anti-HER2 therapy. Therefore, we have sought to identify novel targeted therapies for TNBC. In this study, we investigated the potential of a novel phosphatase, NUDT5, as a potential therapeutic target for TNBC.</p><p><strong>Methods: </strong>The mRNA expression levels of NUDT5 in breast cancers were investigated using TCGA and METABRIC (Curtis) datasets. NUDT5 ablation was achieved through siRNA targeting and NUDT5 inhibition with the small molecule inhibitor TH5427. Xenograft TNBC animal models were employed to assess the effect of NUDT5 inhibition on in vivo tumor growth. Proliferation, death, and DNA replication assays were conducted to investigate the cellular biological effects of NUDT5 loss or inhibition. 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引用次数: 0
摘要
背景:最具侵袭性的乳腺癌是三阴性乳腺癌(TNBC),它缺乏雌激素受体(ER)和孕激素受体(PR)的表达,也没有人类表皮生长因子受体2(HER2)的过度表达。与ER阳性肿瘤可接受激素治疗或HER2阳性肿瘤可接受抗HER2治疗不同,女性TNBC肿瘤患者的治疗选择非常有限。因此,我们一直在寻找治疗 TNBC 的新型靶向疗法。在本研究中,我们研究了新型磷酸酶NUDT5作为TNBC潜在治疗靶点的潜力:方法:利用TCGA和METABRIC(柯蒂斯)数据集研究了NUDT5在乳腺癌中的mRNA表达水平。通过 siRNA 靶向消减 NUDT5,并用小分子抑制剂 TH5427 抑制 NUDT5。采用异种移植 TNBC 动物模型来评估 NUDT5 抑制对体内肿瘤生长的影响。为了研究 NUDT5 缺失或抑制对细胞生物学的影响,还进行了增殖、死亡和 DNA 复制试验。免疫荧光染色法测定了NUDT5缺失后8-氧代鸟嘌呤(8-oxoG)的积累和γH2AX的诱导。通过测量DNA纤维长度评估了NUDT5缺失对复制叉的影响:在这项研究中,我们证实了过表达磷酸酶NUDT5在TNBCs中调节氧化性DNA损伤的重要作用。我们的研究结果表明,NUDT5 的缺失会抑制 TNBC 在体外和体内的生长。这种生长抑制不是由于细胞死亡,而是由于细胞增殖受到抑制。NUDT5 的缺失或抑制导致 DNA 氧化病变 8-oxoG 增加,并引发细胞核中的 DNA 损伤反应。对DNA复制的干扰最终抑制了增殖:结论:NUDT5在防止TNBC细胞的DNA氧化损伤中起着至关重要的作用。结论:NUDT5 在 TNBC 细胞中防止 DNA 氧化损伤方面发挥着关键作用,NUDT5 的缺失或抑制可显著抑制 TNBC 的生长。这些生物学和机理研究为今后的研究奠定了基础,并为开发 NUDT5 抑制剂作为 TNBC 患者的治疗方法提供了可能性。
The novel phosphatase NUDT5 is a critical regulator of triple-negative breast cancer growth.
Background: The most aggressive form of breast cancer is triple-negative breast cancer (TNBC), which lacks expression of the estrogen receptor (ER) and progesterone receptor (PR), and does not have overexpression of the human epidermal growth factor receptor 2 (HER2). Treatment options for women with TNBC tumors are limited, unlike those with ER-positive tumors that can be treated with hormone therapy, or those with HER2-positive tumors that can be treated with anti-HER2 therapy. Therefore, we have sought to identify novel targeted therapies for TNBC. In this study, we investigated the potential of a novel phosphatase, NUDT5, as a potential therapeutic target for TNBC.
Methods: The mRNA expression levels of NUDT5 in breast cancers were investigated using TCGA and METABRIC (Curtis) datasets. NUDT5 ablation was achieved through siRNA targeting and NUDT5 inhibition with the small molecule inhibitor TH5427. Xenograft TNBC animal models were employed to assess the effect of NUDT5 inhibition on in vivo tumor growth. Proliferation, death, and DNA replication assays were conducted to investigate the cellular biological effects of NUDT5 loss or inhibition. The accumulation of 8-oxo-guanine (8-oxoG) and the induction of γH2AX after NUDT5 loss was determined by immunofluorescence staining. The impact of NUDT5 loss on replication fork was assessed by measuring DNA fiber length.
Results: In this study, we demonstrated the significant role of an overexpressed phosphatase, NUDT5, in regulating oxidative DNA damage in TNBCs. Our findings indicate that loss of NUDT5 results in suppressed growth of TNBC both in vitro and in vivo. This growth inhibition is not attributed to cell death, but rather to the suppression of proliferation. The loss or inhibition of NUDT5 led to an increase in the oxidative DNA lesion 8-oxoG, and triggered the DNA damage response in the nucleus. The interference with DNA replication ultimately inhibited proliferation.
Conclusions: NUDT5 plays a crucial role in preventing oxidative DNA damage in TNBC cells. The loss or inhibition of NUDT5 significantly suppresses the growth of TNBCs. These biological and mechanistic studies provide the groundwork for future research and the potential development of NUDT5 inhibitors as a promising therapeutic approach for TNBC patients.
期刊介绍:
Breast Cancer Research, an international, peer-reviewed online journal, publishes original research, reviews, editorials, and reports. It features open-access research articles of exceptional interest across all areas of biology and medicine relevant to breast cancer. This includes normal mammary gland biology, with a special emphasis on the genetic, biochemical, and cellular basis of breast cancer. In addition to basic research, the journal covers preclinical, translational, and clinical studies with a biological basis, including Phase I and Phase II trials.