Saberpour Masoumeh, Maqsoodi Rahimeh, Bakhshi Bita
Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Jalal-Ale-Ahmad Ave, Tehran, 14117-13116, Iran.
BMC Cancer. 2025 Jun 2;25(1):983. doi: 10.1186/s12885-025-14315-5.
Colorectal cancer (CRC) has emerged as a global health concern, as evidenced by its position as the second leading cause of cancer-related mortality. This underscores the necessity for effective disease management strategies. The present study aims to assess the impact of chitosan nanoparticles (CSNP) conjugated with the Lactobacillus acidophilus secretome (CSNP/L.a-sup), on the signaling pathways associated with CRC.
The CSNP/L.a-sup was prepared using an ionic gelation procedure, and its particle size, surface charge, and morphology were evaluated using dynamic light scattering, zeta potential, and scanning electron microscopy. The encapsulation efficiency (EE) and the protein released from CSNP/L.a-sup were assayed using a BCA assay kit. CSNP/L.a-sup toxicity on colon adenocarcinoma (Caco-2) and human dermal fibroblasts (HDF) cells was assessed via the MTT assay. The expression levels of CRC signaling pathway genes were examined using real-time polymerase chain reaction (PCR).
The size of CSNP/L.a-sup was detected at 478.6 ± 219.9 nm, with a surface charge of -8.9 mV. The protein released from CSNP/L.a-sup was observed 76% at pH ~ 6.8 after 48 h, with EE of 74.6%. The viability of Caco-2 and HDF cells against CSNP/L.a-sup was found to be 85.5% and 92.6%, respectively. The uptake of CSNP/L.a-sup by Caco-2 cells occurs in a time-dependent manner, with initial absorption observed within 1 h and substantial internalization achieved after 3 h. CSNP/L.a-sup led to a significant decrease in the expression of β-Catenin, TGF-α, and TGF-β genes, with respective changes of 0.42, 0.79, and 0.16-fold. In contrast, CSNP/L.a-sup led to a significant increase in the expression of PTEN and caspase-9 suppressor genes, with changes of 42.1 and 114.3-fold, respectively. The inhibitory effect of CSNP/L.a-sup on TGF-α gene expression appears to be more closely associated with the CSNP compartment, while the enhancing effect of CSNP/L.a-sup on PTEN gene expression is linked to L.a-sup.
This investigation signifies an inaugural exploration into the potential of a combination therapy comprising secretome of probiotic bacteria and chitosan nanostructures. This approach constitutes a substantial advancement in the field of developing efficacious treatment strategies, offering novel insights into the management of CRC.
结直肠癌（CRC）已成为全球关注的健康问题，其作为癌症相关死亡的第二大主要原因就证明了这一点。这突出了有效疾病管理策略的必要性。本研究旨在评估与嗜酸乳杆菌分泌组结合的壳聚糖纳米颗粒（CSNP/L.a-sup）对与CRC相关的信号通路的影响。
采用离子凝胶法制备CSNP/L.a-sup，并使用动态光散射、zeta电位和扫描电子显微镜评估其粒径、表面电荷和形态。使用BCA检测试剂盒测定CSNP/L.a-sup的包封率（EE）和释放的蛋白质。通过MTT检测评估CSNP/L.a-sup对结肠腺癌（Caco-2）和人皮肤成纤维细胞（HDF）细胞的毒性。使用实时聚合酶链反应（PCR）检测CRC信号通路基因的表达水平。
检测到CSNP/L.a-sup的大小为478.6±219.9nm，表面电荷为-8.9mV。48小时后，在pH约6.8时观察到从CSNP/L.a-sup释放的蛋白质为76%，EE为74.6%。发现Caco-2和HDF细胞对CSNP/L.a-sup的活力分别为85.5%和92.6%。Caco-2细胞对CSNP/L.a-sup的摄取呈时间依赖性，在1小时内观察到初始吸收，3小时后实现大量内化。CSNP/L.a-sup导致β-连环蛋白、TGF-α和TGF-β基因的表达显著降低，变化分别为0.42、0.79和0.16倍。相比之下，CSNP/L.a-sup导致PTEN和caspase-9抑制基因的表达显著增加，变化分别为42.1和114.3倍。CSNP/L.a-sup对TGF-α基因表达的抑制作用似乎与CSNP部分更密切相关，而CSNP/L.a-sup对PTEN基因表达的增强作用与L.a-sup有关。
本研究标志着对益生菌分泌组和壳聚糖纳米结构联合治疗潜力的首次探索。这种方法在开发有效治疗策略领域取得了重大进展，为CRC的管理提供了新的见解。
