中国肿瘤生物治疗杂志2025，Vol.32Issue(7)：681-688,8.DOI:10.3872/j.issn.1007-385x.2025.07.002
CAPN4调控肿瘤干细胞干性在肺腺癌顺铂耐药中的作用机制
Mechanism of CAPN4-mediated cancer stem cell stemness in cisplatin resistance in lung adenocarcinoma
摘要
Abstract
Objective:To investigate the mechanisms by which calpain small subunit 1(CAPN4)regulates cisplatin resistance and cancer stem cell(CSC)stemness in lung adenocarcinoma,and to provide experimental evidence for reversing drug resistance through targeting CSC stemness.Methods:Tissue samples were collected from 10 lung adenocarcinoma patients who underwent surgical resection at Fujian Cancer Hospital from January 2023 to January 2024.Immunohistochemistry(IHC)stining was used to detect the differential expression of CAPN4 in five cisplatin-resistant and five cisplatin-sensitive lung adenocarcinoma tissues,followed with a histological scoring(H-score).CAPN4 gene expression-related survival analysis in lung cancer patients was conducted using The Cancer Genome Atlas(TCGA)database and Gene Expression Profiling Interactive Analysis(GEPIA)platform.Additionally,tissue samples from two cisplatin-resistant and two cisplatin-sensitive lung adenocarcinoma cases were collected to establish lung adenocarcinoma organoid(PDO)models.H-E and IHC staining were used to assess the morphological consistency between PDOs and the primary tumors.CAPN4 gene expression was silenced using lentivirus-mediated shRNA transduction.The expression levels of stem cell markers ALDH1A1,CD133,Nanog,and SOX9 were detected at both the gene and protein levels using quantitative polymerase chain reaction(qPCR)and Western blotting(WB),respectively.The sensitivity of CAPN4-knockdown PDOs to cisplatin was evaluated using the adenosine triphosphate(ATP)assay,and the apoptosis was assessed using the caspase-3 assay.Results:IHC results showed that CAPN4 protein expression was significantly upregulated in cisplatin-resistant lung adenocarcinoma tissues(P<0.05).TCGA cohort analysis revealed that high CAPN4 expression was significantly associated with poor prognosis(reduced OS)in lung adenocarcinoma patients(HR=1.4,P<0.05).PDOs derived from cisplatin-resistant patients exhibited significant upregulation in CAPN4 protein and stemness markers at both gene and protein levels(all P<0.05).Cisplatin sensitivity assays demonstrated that PDOs derived from cisplatin-resistant patients had significantly higher IC50 values than those from cisplatin-sensitive patients(P<0.05).After CAPN4 knockdown,the expression of stem cell makers in PDOs derived from cisplatin-resistant patients were significantly reduced,and their sensitivity to cisplatin was enhanced(P<0.05).Conclusion:Knockdown of CAPN4 reduces stem cell marker expression and enhances cisplatin sensitivity in lung adenocarcinoma PDOs,providing a potential therapeutic target for reversing cisplatin resistance in lung cancer.关键词
肺腺癌/顺铂耐药/肿瘤细胞干性/钙蛋白酶小亚基1Key words
lung adenocarcinoma/cisplatin resistance/tumor cell stemness/calpain small subunit 1(CAPN4)分类
医药卫生引用本文复制引用
张灵玉,刘春江,李秋妹,叶韵斌..CAPN4调控肿瘤干细胞干性在肺腺癌顺铂耐药中的作用机制[J].中国肿瘤生物治疗杂志,2025,32(7):681-688,8.基金项目
福建省自然科学基金(No.2022J05075) （No.2022J05075）
福建省卫生健康中青年骨干人才培养项目(No.2022GGA030) （No.2022GGA030）
福建创新联合基金项目(No.2021Y9198) （No.2021Y9198）
福建肿瘤医院高层次人才培养项目(No.2022YNG03) （No.2022YNG03）
