参芪扶正注射液通过影响 FOXO3 来影响 FBXO22/p53 轴，从而抑制乳酸诱导的 NSCLC 顺铂耐药。

Shenqifuzheng injection inhibits lactic acid-induced cisplatin resistance in NSCLC by affecting FBXO22/p53 axis through FOXO3.

机构信息

College of Integrated Chinese and Western Medical, Liaoning University of Traditional Chinese Medicine, No. 79, Chongshan East Road, Huanggu District, Shenyang City, Liaoning Province, 110847, China.

Pathology Department, Shenyang Key Laboratory for Screening Biomarkers of Tumor Progression and Targeted Therapy of Tumors, Shenyang Medical College, Shenyang City, Liaoning Province, China.

出版信息

Respir Res. 2024 Nov 1;25(1):396. doi: 10.1186/s12931-024-03013-8.

DOI:10.1186/s12931-024-03013-8

PMID:39487426

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11531113/

Abstract

BACKGROUND

Non-small cell lung cancer (NSCLC) accounts for 80% of lung cancers. Cisplatin (DDP)-based combination chemotherapy is the main treatment of NSCLC. Due to resistance to DDP, 5-year overall survival rate of NSCLC patients is very low. Shenqifuzheng injection (SQFZ) is essential for lung cancer progression. However, whether SQFZ plays a role in DDP resistance in NSCLC and its molecular mechanism remains unclear.

METHODS

Levels of FOXO3, FBXO22 and p53 in NSCLC tissues and cells were assessed by RT-qPCR and Western blot. Cell proliferation and apoptosis were analyzed utilizing CCK-8, Colony formation and Flow cytometry assays. Lactate (LA) levels were tested via ELISA. ChIP and Dual luciferase reporter assays validated regulatory relationship between FOXO3 and FBXO22. Immunoprecipitation assay evaluated p53 ubiquitination levels. The subcutaneous tumor model of nude mice was constructed. TUNEL staining detected apoptosis in tissues, and IHC assessed expression of Ki67, FOXO3, FBXO22 and p53.

RESULTS

FOXO3 was decreased, whereas LA and FBXO22 were increased in NSCLC patients. LA led to a higher DDP resistance in A549/DDP cells, while SQFZ reversed this effect by upregulating FOXO3. Furthermore, FBXO22 was a downstream effecter of FOXO3 and FBXO22 affected p53 ubiquitination to reverse the inhibitory effect of SQFZ. We next found SQFZ inhibited LA-induced DDP resistance in NSCLC via FOXO3/FBXO22/p53 axis. Finally, SQFZ regulated LA-mediated DDP resistance in NSCLC nude mice.

CONCLUSION

SQFZ influences LA-induced DDP resistance in NSCLC via FOXO3/FBXO22/p53 pathway, providing a promising agent for NSCLC treatment.

摘要

背景

非小细胞肺癌（NSCLC）占肺癌的 80%。顺铂（DDP）为基础的联合化疗是 NSCLC 的主要治疗方法。由于对 DDP 的耐药性，NSCLC 患者的 5 年总生存率非常低。参芪扶正注射液（SQFZ）是肺癌进展所必需的。然而，SQFZ 是否在 NSCLC 中的 DDP 耐药中发挥作用及其分子机制尚不清楚。

方法

通过 RT-qPCR 和 Western blot 评估 NSCLC 组织和细胞中的 FOXO3、FBXO22 和 p53 水平。利用 CCK-8、集落形成和流式细胞术分析细胞增殖和凋亡。通过 ELISA 测试乳酸（LA）水平。ChIP 和双荧光素酶报告基因检测验证 FOXO3 和 FBXO22 之间的调节关系。免疫沉淀测定评估 p53 泛素化水平。构建裸鼠皮下肿瘤模型。TUNEL 染色检测组织中的细胞凋亡，免疫组化评估 Ki67、FOXO3、FBXO22 和 p53 的表达。

结果

NSCLC 患者中 FOXO3 降低，而 LA 和 FBXO22 升高。LA 导致 A549/DDP 细胞对 DDP 的耐药性增加，而 SQFZ 通过上调 FOXO3 逆转了这种作用。此外，FBXO22 是 FOXO3 的下游效应物，FBXO22 影响 p53 泛素化以逆转 SQFZ 的抑制作用。我们发现 SQFZ 通过 FOXO3/FBXO22/p53 轴抑制 SQFZ 抑制 LA 诱导的 NSCLC 中的 DDP 耐药性。最后，SQFZ 调节 LA 介导的 NSCLC 裸鼠中的 DDP 耐药性。