Inhibition of PI3K/Akt/mTOR Signaling Pathway Suppresses 5-Fluorouracil Resistance in Gastric Cancer.

IF 2.4 4区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular Biotechnology Pub Date : 2024-12-01 Epub Date: 2023-11-24 DOI:10.1007/s12033-023-00966-x

Zhiwei Xing, Yanan Gao, Yaxuan Shi, Ziyu Gao, Caixia Liu

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Abstract

Background: At present, 5-Fluorouracil (5-FU) is a crucial anti-cancer drug and is widely used for the treatment of various carcinomas, including gastric cancer (GC). The resistance of GC cells to 5-FU is still a matter of great concern.

Objective: To illustrate the role of PI3K/Akt/mTOR signaling in regulating the cell cycle progression and migration of 5-FU-resistant GC cells.

Material and methods: After the establishment of drug-resistant GC cell lines, the effects of 5-FU and/or BEZ235 (the dual inhibitor of PI3K and mTOR) on the activity of parental or drug-resistant GC cells were explored. The viability and localization of GC cells (MKN-45 and MKN-74) and their drug-resistant cells (MKN-45/R and MKN-74/R) were assessed using MTT assays and immunofluorescence staining. The impacts of 5-FU and/or BEZ235 on GC cell cycle progression and cell migration were assessed via flow cytometry analyses and wound healing assays, respectively. GC tissues were collected from patients with GC sensitive or refractory to 5-FU chemotherapy. RT-qPCR and western blot were conducted to measure PI3K, AKT, and mTOR levels in GC cells or tissues.

Results: After 5-FU treatment, GC cells displayed 5-FU resistance and the viability of drug-resistant cells (MKN-45/R and MKN-74/R) was higher than that of parental cells (MKN-45 and MKN-74). The IC50 values for MKN-45 and MKN-45/R were 8.93 ug/ml and 140 ug/ml, and the values for MKN-74 and MKN-74/R were 3.93 ug/ml and 114.29 ug/ml. Additionally, the PI3K/Akt/mTOR signaling pathway was activated in drug-resistant GC cells and tumor tissues of patients refractory to 5-FU chemotherapy, as evidenced by high PI3K, Akt, and mTOR levels in MKN-45/R, MKN-74/R, and GC tissues resistant to 5-FU. BEZ235 promoted cell cycle arrest and suppressed the migration of GC cells. Moreover, the combination of BEZ235 and 5-FU led to more effective suppressive influence on cell cycle progression and cell migration relative to the single 5-FU or BEZ235 treatment.

Conclusions: Silencing of the PI3K/Akt/mTOR signaling pathway suppressed the 5-FU resistance of GC cells.

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抑制PI3K/Akt/mTOR信号通路抑制胃癌5-氟尿嘧啶耐药

背景:目前，5-氟尿嘧啶(5-FU)是一种重要的抗癌药物，广泛用于包括胃癌(GC)在内的各种肿瘤的治疗。GC细胞对5-FU的耐药性仍然是一个值得关注的问题。目的:探讨PI3K/Akt/mTOR信号通路在5- fu耐药胃癌细胞周期进程和迁移中的作用。材料和方法:建立耐药GC细胞系后，探讨5-FU和/或BEZ235 (PI3K和mTOR的双重抑制剂)对亲代或耐药GC细胞活性的影响。采用MTT法和免疫荧光染色评价GC细胞(MKN-45和MKN-74)及其耐药细胞(MKN-45/R和MKN-74/R)的活力和定位。5-FU和/或BEZ235对GC细胞周期进展和细胞迁移的影响分别通过流式细胞术分析和伤口愈合试验进行评估。从对5-FU化疗敏感或难治的GC患者中收集GC组织。RT-qPCR和western blot检测GC细胞或组织中PI3K、AKT和mTOR水平。结果:经5-FU处理后，GC细胞表现出5-FU耐药性，耐药细胞(MKN-45/R和MKN-74/R)的活力高于亲本细胞(MKN-45和MKN-74)。MKN-45和MKN-45/R的IC50值分别为8.93 ug/ml和140 ug/ml, MKN-74和MKN-74/R的IC50值分别为3.93 ug/ml和114.29 ug/ml。此外，PI3K/Akt/mTOR信号通路在5-FU化疗难治性胃癌耐药细胞和肿瘤组织中被激活，在5-FU耐药的MKN-45/R、MKN-74/R和GC组织中PI3K、Akt和mTOR水平较高。BEZ235促进细胞周期阻滞，抑制GC细胞迁移。此外，与单独使用5-FU或BEZ235相比，BEZ235和5-FU联合使用对细胞周期进展和细胞迁移的抑制作用更有效。结论:沉默PI3K/Akt/mTOR信号通路可抑制胃癌细胞对5-FU的耐药性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Biotechnology 医学-生化与分子生物学

CiteScore

4.10

自引率

3.80%

发文量

165

审稿时长

6 months

期刊介绍： Molecular Biotechnology publishes original research papers on the application of molecular biology to both basic and applied research in the field of biotechnology. Particular areas of interest include the following: stability and expression of cloned gene products, cell transformation, gene cloning systems and the production of recombinant proteins, protein purification and analysis, transgenic species, developmental biology, mutation analysis, the applications of DNA fingerprinting, RNA interference, and PCR technology, microarray technology, proteomics, mass spectrometry, bioinformatics, plant molecular biology, microbial genetics, gene probes and the diagnosis of disease, pharmaceutical and health care products, therapeutic agents, vaccines, gene targeting, gene therapy, stem cell technology and tissue engineering, antisense technology, protein engineering and enzyme technology, monoclonal antibodies, glycobiology and glycomics, and agricultural biotechnology.