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负载5-氟尿嘧啶的聚乳酸-羟基乙酸共聚物降低了促炎基因IL-9、IL-17A、IL-23和IFN-γ在HT-29结肠癌细胞系中的表达。

5-Fluorouracil-Loaded PLGA Declined Expression of Pro-Inflammatory Genes IL-9, IL-17A, IL-23 and IFN- y; in the HT-29 Colon Cancer Cell Line.

作者信息

Kadhum Kharmeet Basheer, Khalaj-Kondori Mohammad, Hoseinpour Feizi Mohammad Ali, Hajavi Jafar

机构信息

Department of Animal Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.

Department of Microbiology, Faculty of Medicine, Infectious Diseases Research Center, Gonabad University of Medical Science, Gonabad, Iran.

出版信息

Rep Biochem Mol Biol. 2024 Jan;12(4):664-673. doi: 10.61186/rbmb.12.4.664.

Abstract

BACKGROUND

Pro-inflammatory cytokines play critical roles in cancer pathobiology and have been considered potential targets for cancer management and therapy. Understanding the impact of cancer therapeutics such as 5-fluorouracil (5-FU) on their expression might shed light on development of novel combinational therapies. This study aimed to encapsulate 5-FU into PLGA and evaluate their effects on the expression of pro-inflammatory genes , and in the HT-29 cells.

METHODS

PLGA-5-FU NPs were constructed and characterized by Dynamic Light Scattering (DLS) and Atomic Force Microscopy (AFM). The cytotoxicity was evaluated by MTT test and, the IC50 was identified. HT-29 cells were treated with different concentrations of the PLGA-5-FU NPs for 48 hours and, gene expression levels were analyzed by qRT-PCR.

RESULTS

DLS and AFM analysis revealed that the prepared PLGA-5-FU NPs were negatively charged spherical-shaped particles with a mean size of 215.9 ± 43.3 nm. PLGA-5-FU NPs impacted the viability of HT-29 cells in a dose- and time-dependent manner. The qRT-PCR results revealed a dose-dependent decrease in the expression of and genes, and their expressions were significantly different in both 10 and 20 µg/mL treated groups compared to the control. However, although the treatment of HT-29 cells with 20 µg/mL free 5-FU resulted in decreased expression of the studied genes, the differences were not statistically significant compared to the control group.

CONCLUSION

PLGA-5-FU NPs significantly suppressed expression of the and genes, and the encapsulation of 5-FU into PLGA improved considerably impact of the 5-FU on the HT-29 cells.

摘要

背景

促炎细胞因子在癌症病理生物学中发挥关键作用,并被视为癌症管理和治疗的潜在靶点。了解诸如5-氟尿嘧啶(5-FU)等癌症治疗药物对其表达的影响,可能有助于新型联合疗法的开发。本研究旨在将5-FU封装到聚乳酸-羟基乙酸共聚物(PLGA)中,并评估其对HT-29细胞中促炎基因表达的影响。

方法

构建PLGA-5-FU纳米颗粒,并通过动态光散射(DLS)和原子力显微镜(AFM)进行表征。通过MTT试验评估细胞毒性,并确定半数抑制浓度(IC50)。用不同浓度的PLGA-5-FU纳米颗粒处理HT-29细胞48小时,并用实时定量聚合酶链反应(qRT-PCR)分析基因表达水平。

结果

DLS和AFM分析显示,制备的PLGA-5-FU纳米颗粒是带负电荷的球形颗粒,平均尺寸为215.9±43.3纳米。PLGA-5-FU纳米颗粒以剂量和时间依赖性方式影响HT-29细胞的活力。qRT-PCR结果显示,白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)基因的表达呈剂量依赖性下降,与对照组相比,10和20μg/mL处理组的表达均有显著差异。然而,尽管用20μg/mL游离5-FU处理HT-29细胞导致所研究基因的表达下降,但与对照组相比,差异无统计学意义。

结论

PLGA-5-FU纳米颗粒显著抑制IL-6和TNF-α基因的表达,将5-FU封装到PLGA中可显著提高5-FU对HT-29细胞的影响。

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