PO.MCB06.02 · 分子与细胞生物学
zebularine的低甲基化活性抑制多西他赛耐药前列腺癌细胞的迁移
Hypomethylating activity of zebularine suppresses migration in docetaxel-resistant prostate cancer cells
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：前列腺癌（PCa）是男性最常诊断的癌症类型之一。根据美国疾病控制与预防中心（CDC）的数据，每100名男性中，将有13人在其一生中罹患PCa。DNA甲基化已被作为PCa疾病侵袭性和治疗耐药的生物标志物进行研究。尽管PCa被认为是一种生长缓慢的疾病，但它可进展为转移性去势抵抗性前列腺癌（mCRPC）。多西他赛是mCRPC的标准一线化疗药物；然而，近50%的患者会发生耐药。因此，本研究旨在确定改变多西他赛耐药（DR）PCa细胞系的DNA甲基化模式是否会改变关键的细胞功能，包括增殖、迁移以及耐药相关蛋白的表达。我们假设，作为一种DNA甲基转移酶（DNMT）抑制剂，Zebularine会在亲代和DR PCa细胞模型中诱导增殖和迁移的变化。
方法：为评估Zebularine（12.5 – 400 µM）对PCa细胞系（PC-3、DU-145和22Rv1）及其DR对应细胞增殖能力的影响，进行了MTT实验。通过Western Blot评估DNMT1、DNMT3A和MDR1的蛋白表达变化。使用划痕愈合实验评估对细胞迁移能力的影响。
结果：Zebularine处理以浓度依赖方式降低所有细胞系的细胞活力。基于这些结果，选取Zebularine的非毒性浓度（25 µM和100 µM）进行后续实验。除DU145-DR外，Zebularine降低了所有细胞系中DNMT1的表达；DNMT3a或MDR1未见变化。在迁移实验中，与亲代对应细胞相比，Zebularine降低了DR细胞系的迁移能力，尤其是在48和72小时的PC3-DR和DU145-DR细胞中。
结论：Zebularine成功抑制了DNMT1，为其在所用细胞模型中的低甲基化作用提供了证据。此外，它特异性地降低了DR细胞系的细胞迁移。本研究揭示了DNA甲基化在DR细胞系生物学过程调控中的重要性。
资助：由U54 PHSU-MCC资助：U54CA163071 与 U54CA163068；NIH-NIMHD资助 MD007579，以及NIH-NIGMS资助 U54GM133807。
查看英文原文 English abstract
Background: Prostate cancer (PCa) is one of the most common types of cancer diagnosed in men. According to the Centers for Disease Control and Prevention (CDC), for every 100 men, 13 will develop PCa during their lifetime. DNA methylation has been investigated as a biomarker for disease aggressiveness and therapy resistance in PCa. Although PCa is considered a slow-growing disease, it can progress into metastatic castration resistant prostate cancer (mCRPC). Docetaxel is the standard first-line chemotherapy for mCRPC; however, drug resistance occurs in almost 50% of patients. Therefore, this study aimed to determine whether altering DNA methylation patterns in docetaxel-resistant (DR) PCa cell lines modifies key cellular functions, including proliferation, migration, and expression of resistance-related proteins. We hypothesized that Zebularine, a DNA methyltransferase (DNMT) inhibitor, would induce changes proliferation and migration in both parental and DR PCa cell models.
Methods: To evaluate the effects of Zebularine (12.5 - 400 µM) on the proliferative capacity of PCa cell lines (PC-3, DU-145, and 22Rv1) and their DR counterparts, the MTT assay was performed. Protein expression changes in DNMT1, DNMT3A, and MDR1 were evaluated through Western Blot. Effects over cell migration capacity were evaluated using the Wound Healing assay.
Results: Zebularine treatment reduced cell viability in a concentration-dependent manner in all cell lines. Based on these results, non-toxic concentrations of Zebularine (25 µM and 100 µM) were selected for further experiments. Zebularine reduced DNMT1 expression across all cell lines except DU145-DR; no changes were observed for DNMT3a or MDR1. In migration assays, Zebularine reduced the migration capacity of DR cell lines compared with their parental counterparts, particularly in PC3-DR and DU145-DR cells at 48 and 72 hours.
Conclusions: Zebularine led to successful inhibition of DNMT1, providing evidence of its hypomethylating action in the cell models used. Moreover, it led to a reduction in cell migration specifically in DR cell lines. This study sheds light on the importance of DNA methylation in the regulation of biological processes in DR cell lines.
Funding: Sponsored by U54 PHSU-MCC Grants: U54CA163071 & U54CA163068; NIH-NIMHD Grant MD007579, and NIH-NIGMS Grant U54GM133807.
利益披露 Disclosure
G. Castro Morales, None..
D. Gonzalez, None..
G. Alayón, None..
R. Vergne, None..
L. Godoy, None..
G. Ruiz-Deya, None..
C. M. Ortiz-Sanchez, None.