PO.ET04.01 · 实验与分子治疗
利用自递送反义寡核苷酸靶向敲低STEAP2可降低癌症恶性行为
Targeted STEAP2 knockdown with self-delivering antisense oligonucleotide reduces cancer malignant behavior
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景:STEAP(前列腺六次跨膜上皮抗原)家族编码参与铁和铜代谢的金属还原酶,越来越多的证据将金属稳态失调与肿瘤进展联系起来。STEAP2表达增加与多种癌症的发生和进展有关,伴随细胞增殖、迁移增强,但其在癌症中的确切作用仍不明确。本项目的目标是研究靶向STEAP2在肿瘤发生中的意义和预后潜力。
方法:我们的研究考察了一种靶向STEAP2的自递送反义寡核苷酸(ASO)在癌症中的作用。用递增浓度的ASO(0.25、0.5、1和5 μM)处理人肝癌SNU398和前列腺癌PC3细胞92小时,随后评估细胞活力(WST-1)、迁移(Transwell)和非锚定依赖性生长(软琼脂)。在体内研究中,通过将SNU398-luc/GFP细胞注入免疫缺陷小鼠肝脏建立原位异种移植瘤。自第7天起,小鼠接受12剂STEAP2-ASO或乱序对照(5 mg/kg,每两天一次)。通过IVIS成像无创监测肿瘤生长。治疗结束后,采集肿瘤和邻近肝组织。使用Western blot分析在细胞和组织裂解物中评估EMT标志物(Vimentin和SNAIL1)的表达。
结果:ASO介导的STEAP2敲低诱导了STEAP2表达的剂量依赖性降低,导致SNU398和PC3两种细胞系的细胞活力、迁移和非锚定依赖性生长显著减少。此外,STEAP2敲低导致关键EMT标志物(包括Vimentin和SNAIL1)表达降低。体内分析表明,采用靶向STEAP2的ASO治疗使肿瘤体积大幅缩小,同时STEAP2水平下调,凸显了STEAP2抑制与癌症进展抑制之间的明确关联。与体外结果一致,ASO治疗小鼠的肿瘤也显示Vimentin和SNAIL1表达降低,进一步支持EMT减弱。因此,我们的发现强调了STEAP2作为癌症发生和进展的关键调控因子,以及基于ASO的STEAP2沉默在遏制肝癌和前列腺癌进展方面的治疗前景。
结论:总之,这些发现确立了STEAP2作为多种癌症干预的潜在生物标志物和治疗靶点。
查看英文原文 English abstract
Background: The STEAP (Six-Transmembrane Epithelial Antigen of Prostate) family encodes metalloreductases involved in iron and copper metabolism, with increasing evidence linking dysregulated metal homeostasis to tumor progression. Increased STEAP2 expression has been implicated in the development and progression of several cancers, with enhanced cellular proliferation, migration, but its precise role in cancer remains obscure. The goal of this project is to investigate the significance and prognostic potential of targeting STEAP2 in oncogenesis.
Methods: Our study investigated a self-delivering antisense oligonucleotide (ASO) targeting STEAP2 in cancer. Human liver cancer SNU398 and prostate cancer PC3 cells were treated with increasing ASO concentrations (0.25, 0.5, 1 and 5 μM) for 92 hours, followed by assessment of cell viability (WST-1), migration (transwell), and anchorage-independent growth (soft agar). For the in vivo study, orthotopic xenografts were created by injecting SNU398-luc/GFP cells into the livers of immunodeficient mice. Beginning on day 7, mice received 12 doses of the STEAP2-ASO or scrambled control (5 mg/kg, every two days). Tumor growth was monitored non-invasively via IVIS imaging. Upon treatment completion, tumors and adjacent liver tissues were harvested. Expression of EMT markers (Vimentin and SNAIL1) was evaluated in cell and tissue lysates using western blot analysis.
Results: ASO mediated KD of STEAP2 induced a dose dependent reduction in STEAP2 expression, leading to significantly reduced cell viability, migration, and anchorage-independent growth in both SNU398 and PC3 cell lines. In addition, STEAP2 knockdown resulted in decreased expression of key EMT markers, including Vimentin and SNAIL1. In vivo analysis demonstrated that treatment with STEAP2 targeting ASOs resulted in a substantial decrease in tumor size, together with downregulation of STEAP2 levels, highlighting a clear association between STEAP2 suppression and inhibition of cancer progression. Consistent with the in vitro findings, tumors from ASO-treated mice also showed reduced Vimentin and SNAIL1 expression, further supporting EMT attenuation. Thus, our findings emphasize STEAP2 as an essential regulator of cancer development and progression as well as the therapeutic promise of the ASO-based STEAP2 silencing for abrogating the progression of liver and prostate cancer.
Conclusion: Overall, these findings establish STEAP2 as a potential biomarker and therapeutic target for the intervention of various cancers.
利益披露 Disclosure
N. K. Panchal, None..
A. Bernal, None..
B. Barre, None..
F. Cigarroa, None..
L. Sun, None.