G 蛋白信号调节器 20 通过抑制化脓过程提高非小细胞肺癌细胞的放射抗性

Q1 Health Professions

Radiation Medicine and Protection Pub Date : 2024-05-23 DOI:10.1016/j.radmp.2024.05.008

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引用次数: 0

摘要

目的研究G蛋白信号转导调节因子20（RGS20）在非小细胞肺癌（NSCLC）放射抗性中的潜在作用。方法从癌症基因组图谱（TCGA）中收集了35例接受放疗的肺腺癌（LUAD）患者，并根据临床预后将其分为放射敏感组（16例）和放射抗性组（19例）。基因表达谱交互分析（GEPIA）数据库分析了RGS20的表达和预后。用 6 Gy X 射线照射 A549 细胞 10 次，构建抗放射细胞系（A549R）。细胞存活率通过集落形成试验测定。结果通过分析 TCGA-LUAD 数据集中放射敏感和放射耐受个体的数据，共鉴定出 2,181 个差异表达基因（DEGs）。这些 DEGs 在 Reactome 数据库分析的 G alpha (z) 信号事件中富集。在 DEGs 中，RGS20 表现出明显的上调，其较高的表达预示着 LUAD 患者的不良预后。在体外，RGS20蛋白在A549细胞中的表达因辐照而增加，而在A549R细胞中，无论辐照与否，RGS20蛋白都保持在很高的水平。辐照后，A549 细胞中热休克相关蛋白的表达明显增加（P ＜ 0.05），而 A549R 细胞中则无明显变化。用 LDC7559 处理可明显减少 LDH 的释放（P ＜ 0.01），并提高辐照 A549 细胞的存活率（P ＜ 0.01）。此外，敲除 A549R 细胞中的 RGS20 基因可明显增加 LDH 的释放（P ＜ 0.001）并增强辐射敏感性（P ＜ 0.01），而服用 LDC7559 则可逆转 siRGS20 导致的 LDH 释放（P ＜ 0.01）和辐射诱导的细胞死亡（P ＜ 0.05）。同时，在转染siRGS20的A549和A549R细胞中观察到GSDMD-NT表达水平升高（P＜0.05）。

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Regulator of G protein signaling 20 contributes to radioresistance of non-small cell lung cancer cells by suppressing pyroptosis

Objective

To investigate the potential role of the regulator of G protein signaling 20 (RGS20) in radioresistance of non-small cell lung cancer (NSCLC).

Methods

A total of 35 lung adenocarcinoma (LUAD) patients from The Cancer Genome Atlas (TCGA), who underwent radiotherapy, were enrolled and divided into radiosensitive (n = 16) and radioresistant (n = 19) groups based on clinical prognosis. The expression and prognosis of RGS20 were analyzed by Gene Expression Profiling Interactive Analysis (GEPIA) database. A radioresistant cell line (A549R) was constructed by irradiating A549 cells with 6 Gy X-rays for 10 fractions. Cell survival was measured by colony formation assay. The regulatory effect of RGS20 on pyroptosis were verified by LDH release and Western blot assay, and the underlying mechanism was investigated by transfecting RGS20 siRNA and applying a GSDMD inhibitor).

Results

A total of 2,181 differentially expressed genes (DEGs) were identified by analyzing the data of radiosensitive and radioresistant individuals from the TCGA-LUAD dataset. These DEGs were enriched in G alpha (z) signalling events analyzed by Reactome database. RGS20 exhibited significant upregulation among the DEGs, and its higher expression predicted poor prognosis in LUAD patients. In vitro, the expression of RGS20 protein was increased by irradiation in A549 cells, whereas it remained at much high levels in A549R cells regardless of irradiation. After irradiation, the expressions of pyroptosis-related proteins were significantly increased in A549 cells (P ＜ 0.05), with no significant changes were observed in A549R cells. Treatment with LDC7559 significantly reduced LDH release (P ＜ 0.01) and improved the survival rate of irradiated A549 cells (P ＜ 0.01). Furthermore, knockdown of RGS20 gene in A549R cells significantly increased LDH release (P ＜ 0.001) and enhanced radiosensitivity (P ＜ 0.01), while LDC7559 administration reversed LDH release (P ＜0.01) and radiation-induced cell death increased by siRGS20 (P ＜0.05). Meantime, the increased expression level of GSDMD-NT was observed in A549 and A549R cells transfected with siRGS20 (P ＜ 0.05).