Eksi Orhan Burak, Guler Ahsen, Akdeniz Munevver, Atalay Pinar, Hamurcu Zuhal, Aydin Omer
ERNAM-Nanotechnology Research and Application Center, Erciyes University, Kayseri, 38039, Turkey.
NanoThera Lab, ERFARMA-Drug Application and Research Center, Erciyes University, Kayseri, 38039, Turkey.
Drug Deliv Transl Res. 2025 Apr 23. doi: 10.1007/s13346-025-01860-6.
Breast cancer is the most common cancer among women, with approximately 2.3 million new cases globally. Triple-negative breast cancer (TNBC) is an aggressive subtype characterized by the lack of estrogen receptor (ER), progesterone receptor (PR), and HER2 expression, making it unresponsive to traditional therapies. Eukaryotic Elongation Factor 2 Kinase (eEF2K) is overexpressed in TNBC, promoting cell survival by inhibiting apoptosis through phosphorylation of eEF2. Recently, eEF2K has been targeted for cancer therapy, and siRNA-based gene therapy has emerged as an effective approach to silence overexpressed genes. However, siRNA delivery is challenging due to its instability and susceptibility to degradation. In this study, we developed a novel hybrid nanoparticle (HNP) using a Layer-by-Layer (LbL) method for siRNA delivery targeting eEF2K in TNBC. The HNPs consist of a silver nanoparticle (AgNP) core, coated with poly (allylamine hydrochloride) (PAH) and poly(styrene sulfonate) (PSS), and loaded with eEF2K-siRNA and quercetin (QU), a chemotherapeutic agent, in separate layers. The nanoparticles also incorporated 4-ATP molecules for Raman traceability. In vitro experiments on TNBC cell lines (MDA-MB-231, BT-549, 4T1) showed that the combination therapy of eEF2K-siRNA and QU reduced cell viability, inhibited colony formation, and suppressed cell migration. At high 120 nM of siRNA concentration, 3D spheroid disintegration, activation of apoptotic pathways, and eventual necrotic cell death were observed. The results demonstrate that the developed HNPs are non-toxic, effective, and offer potential as a theranostic platform for TNBC treatment.
乳腺癌是女性中最常见的癌症，全球每年约有230万新发病例。三阴性乳腺癌（TNBC）是一种侵袭性亚型，其特征是缺乏雌激素受体（ER）、孕激素受体（PR）和HER2表达，这使得它对传统疗法无反应。真核延伸因子2激酶（eEF2K）在TNBC中过度表达，通过磷酸化eEF2抑制细胞凋亡来促进细胞存活。最近，eEF2K已成为癌症治疗的靶点，基于小干扰RNA（siRNA）的基因治疗已成为沉默过度表达基因的有效方法。然而，由于siRNA的不稳定性和易降解性，其递送具有挑战性。在本研究中，我们使用层层（LbL）方法开发了一种新型混合纳米颗粒（HNP），用于在TNBC中靶向递送针对eEF2K的siRNA。HNP由银纳米颗粒（AgNP）核心组成，表面涂覆有聚（烯丙胺盐酸盐）（PAH）和聚（苯乙烯磺酸盐）（PSS），并在不同层中负载了eEF2K-siRNA和化疗药物槲皮素（QU）。纳米颗粒还掺入了4-ATP分子用于拉曼可追溯性。对TNBC细胞系（MDA-MB-231、BT-549、4T1）的体外实验表明，eEF2K-siRNA和QU的联合治疗降低了细胞活力，抑制了集落形成，并抑制了细胞迁移。在siRNA浓度为120 nM时，观察到三维球体解体、凋亡途径激活以及最终的坏死性细胞死亡。结果表明，所开发的HNP无毒、有效，有望成为TNBC治疗的治疗诊断平台。
