Methionine restriction promotes cisplatin sensitivity of gastric cancer resistant cells by down-regulating circ-CDK13 level

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research Pub Date : 2024-11-01 DOI:10.1016/j.yexcr.2024.114315

Lin Xin , Yong-Hui Zou , Chen-Xi Liu , Hao Lu , Luo-Jun Fan , He-Song Xu , Qi Zhou , Jiang Liu , Zhen- Qi Yue , Jin-Heng Gan

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Abstract

Background

Methionine restriction (MR) is a research direction in the treatment of gastric cancer (GC). The aim of this study was to investigate the molecular mechanism of MR on enhancing cisplatin (DDP) sensitivity of drug-resistant GC cells.

Methods

Twenty pairs of GC tissues and adjacent normal gastric mucosa tissues were collected. DDP-resistant cell lines (KATO/DDP and MKN45/DDP), mouse model of GC and GC patient-derived organoid (PDO) models were established. Lentivirus-mediated METase overexpression was used for MR. Cell viability and apoptosis were detected by MTT assay and flow cytometry. Western blotting was used to detect multi-drug resistance-1 (MDR1), MDR-associated protein 1 (MRP1) eukaryotic initiation factor 4A-Ⅲ (EIF4A3), and METase protein expressions. The levels of circRNAs were detected by qRT-PCR. Tumor volume and weight were measured. The proliferation of tumor cells was detected by immunohistochemical staining.

Results

The differentially expressed circRNAs of GC were screened in Gene Expression Omnibus database. MR in KATO/DDP and MKN45/DDP cells significantly down-regulated circ-CDK13 level. Overexpression of circ-CDK13 significantly inhibited apoptosis of sensitive cells (KATO III and MKN45). Interference with circ-CDK13 significantly promoted apoptosis of drug-resistant cells (KATO/DDP and MKN45/DDP). MR enhanced the DDP sensitivity of GC resistant cells, GC PDO and GC mice by down-regulating circ-CDK13. EIF4A3 binds to the downstream flanking sequence of circ-CDK13, and interference with EIF4A3 reduces circ-CDK13 levels, but does not affect CDK13. The expressions of circ-CDK13 and EIF4A3 in GC clinical samples were increased and positively correlated. Simultaneously overexpression of METase and EIF4A3 in resistant cells inhibited apoptosis, and further interference with circ-CDK13 reversed this effect.

Conclusion

MR inhibits circ-CDK13 level by down-regulating EIF4A3, thereby increasing the sensitivity of GC drug-resistant cells to DDP.

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限制蛋氨酸可通过下调循环 CDK13 水平促进胃癌耐药细胞对顺铂的敏感性

背景：甲硫氨酸限制（MR）是治疗胃癌（GC）的一个研究方向。本研究的目的是探讨 MR 增强耐药 GC 细胞对顺铂 (DDP) 敏感性的分子机制：方法：收集 20 对 GC 组织和邻近正常胃黏膜组织。建立了耐 DDP 细胞系（KATO/DDP 和 MKN45/DDP）、GC 小鼠模型和 GC 患者衍生类器官（PDO）模型。慢病毒介导的 METase 过表达被用于 MR。通过 MTT 检测法和流式细胞术检测细胞活力和凋亡。用 Western 印迹法检测多药耐药性-1（MDR1）、MDR 相关蛋白 1（MRP1）、真核起始因子 4A-Ⅲ （EIF4A3）和 METase 蛋白的表达。通过 qRT-PCR 检测 circRNAs 的水平。测量肿瘤体积和重量。免疫组化染色检测肿瘤细胞的增殖情况：结果：基因表达总库（Gene Expression Omnibus）数据库筛选出 GC 的差异表达 circRNAs。KATO/DDP和MKN45/DDP细胞中的MR显著下调circ-CDK13水平。过表达 circ-CDK13 能明显抑制敏感细胞（KATO III 和 MKN45）的凋亡。干扰 circ-CDK13 能明显促进耐药细胞（KATO/DDP 和 MKN45/DDP）的凋亡。MR通过下调circ-CDK13增强了GC耐药细胞、GC PDO和GC小鼠对DDP的敏感性。EIF4A3 与 circ-CDK13 的下游侧翼序列结合，干扰 EIF4A3 会降低 circ-CDK13 的水平，但不会影响 CDK13。在 GC 临床样本中，circ-CDK13 和 EIF4A3 的表达量增加并呈正相关。在耐药细胞中同时过表达 METase 和 EIF4A3 可抑制细胞凋亡，而进一步干扰 circ-CDK13 可逆转这种效应：结论：MR通过下调EIF4A3抑制circ-CDK13水平，从而提高了GC耐药细胞对DDP的敏感性。

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

Experimental cell research 医学-细胞生物学

CiteScore

7.20

自引率

0.00%

发文量

295

审稿时长

30 days

期刊介绍： Our scope includes but is not limited to areas such as: Chromosome biology; Chromatin and epigenetics; DNA repair; Gene regulation; Nuclear import-export; RNA processing; Non-coding RNAs; Organelle biology; The cytoskeleton; Intracellular trafficking; Cell-cell and cell-matrix interactions; Cell motility and migration; Cell proliferation; Cellular differentiation; Signal transduction; Programmed cell death.