HSPA4诱导内质网应激激活β-catenin信号通路促进肺腺癌进展的机制研究OA
HSPA4 promotes progression of lung adenocarcinoma by inducing endo-plasmic reticulum stress and activating β-catenin signaling pathway
目的:通过检测热休克蛋白A4(heat shock protein A4,HSPA4)在肺腺癌中的表达,探讨其通过诱导内质网应激(endoplasmic reticulum stress,ERS)并激活Wnt/β-catenin信号通路促进肺腺癌的进展机制.方法:本研究利用TCGA数据库分析HSPA4在肺腺癌中的表达及其与预后的关系,并收集了2020年1月至2023年12月在皖南医学院第一附属医院确诊的60例肺腺癌患者的血液和组织样本,以及40例体检健康对照者的血液样本.通过免疫组化、Western blot和酶联免疫吸附方法检测HSPA4水平并分析其与肺腺癌预后的关系.在体外实验中,使用HSPA4特异性小干扰RNA(small interfering RNA,siRNA)和质粒pcDNA对肺腺癌细胞进行HSPA4的敲减和过表达,使用EdU染色评估细胞增殖能力,同时检测HSPA4、HSPA5、增殖细胞核抗原(proliferating cell nuclear anti-gen,PCNA)、β-catenin及其下游因子cyclin D1、c-Myc的表达水平.结果:数据库资料分析显示HSPA4在肺腺癌患者中表达显著升高(P<0.01),HSPA4表达影响肺腺癌患者预后(P<0.01).临床样本中,免疫组化结果显示肺腺癌患者癌组织HSPA4水平较癌旁正常组织显著升高(P<0.01),肺腺癌患者血清中HSPA4水平较对照组显著升高(P<0.01).体外实验中,Western blot检测显示,HSPA4在肺腺癌细胞系中表达水平存在差异,其中A549和HCC827中HSPA4表达水平显著升高(P<0.01),H1299中HSPA4表达水平显著降低(P<0.01),选取H1299和A549进行后续研究.敲减A549细胞中HSPA4可抑制细胞增殖,下调PCNA表达,并降低HSPA5、β-catenin及其下游cyclin D1、c-Myc的表达.在H1299细胞中过表达HSPA4则促进细胞增殖及PCNA表达,同时上调HSPA5、β-catenin、cyclin D1和c-Myc;该效应可被ERS抑制剂4-PBA部分逆转.结论:HSPA4通过诱导ERS并激活β-catenin信号通路促进肺腺癌的进展.
AIM:This study aims to investigate the role of heat shock protein A4(HSPA4)in promoting the progression of lung adenocarcinoma by inducing endoplasmic reticulum stress(ERS)and activating the Wnt/β-catenin sig-naling pathway,through the detection of its expression in lung adenocarcinoma.METHODS:The expression of HSPA4 in lung adenocarcinoma and its correlation with prognosis were analyzed using the TCGA database.Tissue and blood sam-ples were collected from 60 patients diagnosed with lung adenocarcinoma at The First Affiliated Hospital of Wannan Medi-cal College between January 2020 and December 2023,alongside blood samples from 40 healthy controls.Immunohisto-chemistry,Western blot,and enzyme-linked immunosorbent assay were utilized to detect HSPA4 levels and analyze their relationship with prognosis.In vitro,HSPA4-specific siRNA and pcDNA were employed to knock down or overexpress HS-PA4 in lung adenocarcinoma cells.Cell proliferation was assessed using EdU staining,and the expression levels of HS-PA4,HSPA5,proliferating cell nuclear antigen(PCNA),β-catenin,and its downstream factors,cyclin D1 and c-Myc,were examined.RESULTS:Database analysis revealed significantly elevated expression of HSPA4 in lung adenocarcino-ma patients(P<0.01),which correlated with prognosis(P<0.01).In clinical samples,immunohistochemistry demon-strated significantly higher levels of HSPA4 in cancer tissues compared to adjacent normal tissues(P<0.01).Additional-ly,serum HSPA4 levels were significantly increased in patients compared to controls(P<0.01).In vitro studies using Western blot analysis revealed differential HSPA4 expression across lung adenocarcinoma cell lines,with significantly higher levels observed in A549 and HCC827 cells(P<0.01)and lower levels in H1299 cells(P<0.01).A549 and H1299 cell lines were selected for subsequent experiments.Knockdown of HSPA4 in A549 cells suppressed cell prolifera-tion,downregulated PCNA expression,and reduced the levels of HSPA5,β-catenin,cyclin D1,and c-Myc.Conversely,overexpression of HSPA4 in H1299 cells promoted cell proliferation and PCNA expression while upregulating HSPA5,β-catenin,cyclin D1,and c-Myc.These effects were partially reversed by the endoplasmic reticulum stress(ERS)inhibi-tor 4-PBA.CONCLUSION:HSPA4 promotes lung adenocarcinoma progression by inducing ERS and activating the β-catenin signaling pathway.
秦立龙;王晓彤;汪丽静;臧蕾蕾;程玉生;徐雯
皖南医学院第一附属医院呼吸与危重症医学科,安徽 芜湖 241000皖南医学院第一附属医院心功能科,安徽 芜湖 241000皖南医学院第一附属医院呼吸与危重症医学科,安徽 芜湖 241000皖南医学院第一附属医院呼吸与危重症医学科,安徽 芜湖 241000皖南医学院第一附属医院呼吸与危重症医学科,安徽 芜湖 241000安徽医科大学附属滁州医院(滁州市第一人民医院)呼吸与危重症医学科,安徽 滁州 239001
医药卫生
肺腺癌热休克蛋白A4内质网应激β-catenin
lung adenocarcinomaheat shock protein A4endoplasmic reticulum stressβ-catenin
《中国病理生理杂志》 2026 (5)
875-883,9
安徽省卫健委科研重大项目(No.AHWJ2023A10132)
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