基于PI3K/AKT信号通路探讨PRDX4对食管鳞状细胞癌细胞增殖和凋亡的影响

Exploring the effect of PRDX4 on proliferation and apoptosis of esophageal squamous cell carcinoma cells based on the PI3K/AKT signaling pathway

  • 摘要:
    目的 分析过氧化物还原酶4(peroxiredoxin 4,PRDX4)对食管鳞状细胞癌(esophageal carcinoma,ESCC)细胞增殖和凋亡能力的调控作用及相关蛋白表达。
    方法 结合UALCAN、GEPIA和TCGA数据库预测PRDX4在ESCC中的表达及与病理特征及预后的关系。选取2010年8月至2023年8月潍坊医学院附属医院行食管癌(esophageal carcinoma,EC)根治术治疗的60例ESCC患者的癌组织及癌旁组织为研究样本,分析组织样本中的PRDX4的表达水平。通过实时荧光定量PCR和免疫印迹法分析ESCC细胞中PRDX4的mRNA和相关信号分子蛋白的表达水平;此外,结合CCK-8实验、流式细胞术实验分析PRDX4对细胞的增殖和凋亡活动的影响。最终将体外实验结果通过裸鼠皮下瘤模型进行验证。
    结果 GEPIA和UALCAN数据库的数据表明,PRDX4在ESCC中表达异常增高,且与病理分期、分级和患者的生存率等有关。对ESCC细胞系PRDX4进行敲低和过表达后,PRDX4、p-PI3K、p-AKT、Cyclin D1、Survivin蛋白表达分别表达降低和增高,细胞增殖和凋亡能力均受正相关调控;相较于sh-NC组、sh-PRDX4组裸鼠肿瘤的体积和质量降低。
    结论 PRDX4可基于P13K/AKT信号通路的激活对ESCC细胞的增殖和凋亡能力进行调控。

     

    Abstract:
    Objective Studying the effect of PRDX4 on esophageal squamous cell carcinoma cells (esophageal carcinoma, ESCC) proliferation and apoptosis as well as its potential mechanism.
    Methods The University of Alabama at Birmingham cancer data analysis portal (UALCAN), gene expression profiling interactive analysis (GEPIA) and the Cancer Genome Atlas (TCGA) databases were used to predict PRDX4 expression in ESCC and its relationship with pathological features and prognosis. The cancer and adjacent tissues of 60 patients with ESCC who underwent radical resection in the Affiliated Hospital of Weifang Medical College from August 2010 to August 2023 were selected as research samples. The expression level of PRDX4 in the patients was detected by immunohistochemistry (IHC). The extracted cancer and adjacent tissues were homogenized to analyze its mRNA expression. The expression levels of PRDX4 mRNA and related signaling proteins in ESCC cells were analyzed by real-time quantitative PCR and Western blot. Cell Counting Kit-8 (CCK-8) assay and flow cytometry were used to analyze the effect of PRDX4 on cell proliferation and apoptosis. Finally, a subcutaneous tumor model in nude mice was constructed to validate the in vitro experimental results.
    Results The data from the GEPIA and UALCAN showed that PRDX4 expression was abnormally increased and related to the pathology stage, grade, and survival rate of patients. After knockdown and overexpression of PRDX4 in an ESCC cell line, the expression of PRDX4, phos-phosphatidylinositol 3-kinase (p-PI3K), phos-protein kinase B (p-AKT), cyclinD1, and survivin protein decreased and increased, respectively; cell proliferation and apoptosis were positively regulated. Compared with the sh-NC group, tumor volume and weight in the sh-PRDX4 group were decreased.
    Conclusions PRDX4 regulates the proliferation and apoptosis of ESCC cells by activating the PI3K/AKT signaling pathway.

     

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