抑制 PI3K/Akt/mTOR 信号通路可增强胃癌对 5-氟尿嘧啶的敏感性。
Inhibition of PI3K/Akt/mTOR Signaling Pathway Suppresses 5-Fluorouracil Resistance in Gastric Cancer.
机构信息
Department of Oncology, Affiliated Hospital of Inner Mongolia Medical University, No.1, Tongdao North Road, Huimin District, Hohhot, 010010, Inner Mongolia Autonomous Region, China.
Graduate School of Inner Mongolia Medical University, Hohhot, 010010, Inner Mongolia Autonomous Region, China.
出版信息
Mol Biotechnol. 2024 Dec;66(12):3640-3654. doi: 10.1007/s12033-023-00966-x. Epub 2023 Nov 24.
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.
背景
目前，5-氟尿嘧啶（5-FU）是一种重要的抗癌药物，广泛用于治疗各种癌症，包括胃癌（GC）。GC 细胞对 5-FU 的耐药性仍然是一个备受关注的问题。
目的
阐述 PI3K/Akt/mTOR 信号通路在调节 5-FU 耐药 GC 细胞周期进程和迁移中的作用。
材料和方法
在建立耐药 GC 细胞系后，研究了 5-FU 和/或 BEZ235（PI3K 和 mTOR 的双重抑制剂）对亲本或耐药 GC 细胞活性的影响。通过 MTT 检测和免疫荧光染色评估 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。通过流式细胞术分析和划痕愈合试验分别评估 5-FU 和/或 BEZ235 对 GC 细胞周期进程和细胞迁移的影响。收集对 5-FU 化疗敏感或耐药的 GC 患者的 GC 组织。通过 RT-qPCR 和 Western blot 测量 GC 细胞或组织中的 PI3K、AKT 和 mTOR 水平。
结果
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 8.93ug/ml 和 140ug/ml，MKN-74 和 MKN-74/R 的 IC50 值分别为 3.93ug/ml 和 114.29ug/ml。此外，PI3K/Akt/mTOR 信号通路在耐药 GC 细胞和对 5-FU 化疗耐药的患者肿瘤组织中被激活，MKN-45/R、MKN-74/R 和对 5-FU 耐药的 GC 组织中 PI3K、Akt 和 mTOR 水平升高。BEZ235 促进 GC 细胞周期停滞并抑制细胞迁移。此外，与单独使用 5-FU 或 BEZ235 相比，BEZ235 与 5-FU 的联合使用对细胞周期进程和细胞迁移具有更有效的抑制作用。
结论
沉默 PI3K/Akt/mTOR 信号通路抑制了 GC 细胞对 5-FU 的耐药性。
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