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

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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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Data Availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

The work was supported by Inner Mongolia Medical University “Zhiyuan Talent Program” (No. ZY0202030), Health Commission of Inner Mongolia Autonomous Region Health Medical and Health Science and Technology Project 2022 (No. 202202196), and the Affiliated Hospital Inner Mongolia Medical University Scientific Research Project (No. NYFY YB 005).

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Authors and Affiliations

  1. Department of Oncology, Affiliated Hospital of Inner Mongolia Medical University, No.1, Tongdao North Road, Huimin District, Hohhot, 010010, Inner Mongolia Autonomous Region, China

    Zhiwei Xing, Ziyu Gao & Caixia Liu

  2. Graduate School of Inner Mongolia Medical University, Hohhot, 010010, Inner Mongolia Autonomous Region, China

    Yanan Gao & Yaxuan Shi

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  1. Zhiwei Xing
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  2. Yanan Gao
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Contributions

ZX and CL contributed to Study Conception, Data Analysis, and Draft Manuscript Preparation. YG and YS contributed to Data Collection and Perform Experiment. ZG contributed to Data Collection and Experimental guidance.

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Correspondence to Caixia Liu.

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The authors declare that they have no competing interests.

Ethical Approval

The Ethics Committee of Affiliated Hospital of Inner Mongolia Medical University approved this study (No. YKD2017034, March 7, 2017) and all experiments were conducted in line with the Declaration of Helsinki.

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Xing, Z., Gao, Y., Shi, Y. et al. Inhibition of PI3K/Akt/mTOR Signaling Pathway Suppresses 5-Fluorouracil Resistance in Gastric Cancer. Mol Biotechnol (2023). https://doi.org/10.1007/s12033-023-00966-x

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