Khodadadi Ghazale, Saberpour Masoumeh, Bakhshi Bita, Minaeian Sara
Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Jalal-Ale-Ahmad Ave., Tehran, 14117-13116 Iran.
Department of Influenza and Other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran.
Cytotechnology. 2025 Jun;77(3):93. doi: 10.1007/s10616-025-00756-0. Epub 2025 Apr 28.
Colorectal cancer (CRC) remains as a major health problem with high lethality rate in the world. The innovate therapeutic strategies are essential in CRC management. The purpose of this research was to evaluate the effect of chitosan nanoparticle containing culture supernatant (CNP/Cj-sup) on genes involved in CRC signaling pathways. CNP/Cj-sup was fabricated via ionotropic gelation method. Dynamic light scattering and transmission electron microscopy (TEM) techniques were employed to characterize of the CNP/Cj-sup including the electrical charge, size distribution, and morphological properties. The loaded protein, released protein, and entrapment efficacy (EE) were assayed utilizing a BCA assay kit. After the evaluation of the viability of Caco-2 (colon adenocarcinoma) and HDF (human dermal fibroblasts) cells against CNP/Cj-sup by MTT assay, subsequently anti-tumor effect of CNP/Cj-sup on genes associated with CRC signaling pathways was assessed via real-time PCR method. The size dispersion of CNP/Cj-sup was 400.6 ± 24.4 nm with an electrical charge of + 4.5 mV. The loaded protein was calculated 1100 µg. The release rate of protein from CNP/Cj-sup was 78% at pH of 6.8 after 48 h, with EE of 74.62%. The viability of Caco-2 and HDF cells against CNP/Cj-sup (1100 µg + 0.05%) was measured 75.8 and 96.5%, respectively after 48 h. CNP/Cj-sup exhibited the highest efficacy in inhibiting the expression of oncogenes , and by to 0.06, 0.34, 0.14, 0.13, 0.08, and 0.14-fold ( value < .0001). Moreover, it led to a significant increase in the expression of the suppressor genes and by to 55.7 and 1.8- fold ( value < .0001). CNP/Cj-sup demonstrated the highest efficiency in suppressing and enhancing compared to CNP and Cj-sup. In conclusion, CNP/Cj-sup as an innovative potential anticancer agent, with the ability to modulate genes involved in CRC progression, represents a promising approach to CRC treatment.
The effect of CNP/Cj-sup on different colorectal cancer signaling pathways.
结直肠癌（CRC）仍是全球致死率较高的主要健康问题。创新的治疗策略对于CRC的管理至关重要。本研究旨在评估含壳聚糖纳米颗粒的培养上清液（CNP/Cj-sup）对CRC信号通路相关基因的影响。通过离子凝胶法制备CNP/Cj-sup。采用动态光散射和透射电子显微镜（TEM）技术对CNP/Cj-sup的电荷、尺寸分布和形态特性进行表征。使用BCA检测试剂盒测定负载蛋白、释放蛋白和包封率（EE）。通过MTT法评估Caco-2（结肠腺癌）和HDF（人皮肤成纤维细胞）细胞对CNP/Cj-sup的活力后，随后通过实时PCR法评估CNP/Cj-sup对CRC信号通路相关基因的抗肿瘤作用。CNP/Cj-sup的尺寸分散度为400.6±24.4nm，电荷为+4.5mV。负载蛋白计算为1100μg。48小时后，在pH值为6.8时，CNP/Cj-sup的蛋白释放率为78%，包封率为74.62%。48小时后，Caco-2和HDF细胞对CNP/Cj-sup（1100μg+0.05%）的活力分别测定为75.8%和96.5%。CNP/Cj-sup在抑制癌基因表达方面表现出最高的功效，分别降低至0.06、0.34、0.14、0.13、0.08和0.14倍（P值<0.0001）。此外，它导致抑癌基因和的表达显著增加，分别增加至55.7倍和1.8倍（P值<0.0001）。与CNP和Cj-sup相比，CNP/Cj-sup在抑制和增强方面表现出最高的效率。总之，CNP/Cj-sup作为一种具有创新潜力的抗癌剂，具有调节参与CRC进展的基因的能力，代表了一种有前景的CRC治疗方法。
CNP/Cj-sup对不同结直肠癌信号通路的影响。
