安徽医科大学学报

目的 探讨无精子症缺失基因相关蛋白2(DAZAP2)在乳腺癌中的表达及其对肿瘤生物学行为的影响。方法 通过TCGA数据库分析DAZAP2的表达水平与患者生存之间的关系。采用免疫组化方法检测DAZAP2在乳腺癌和癌旁组织中的表达水平并分析其与临床病理特征的关系，使用Kaplan-Meier对不同DAZAP2表达水平患者的总生存期（OS）及无复发生存期（RFS）进行分析。分别构建敲低和过表达DAZAP2的乳腺癌稳转细胞系，通过CCK-8与克隆形成实验评估乳腺癌细胞增殖能力的变化，通过细胞划痕和Transwell实验评估乳腺癌细胞迁移和侵袭能力的变化。结果 生物信息学分析和免疫组化结果显示，DAZAP2在乳腺癌组织中的表达水平低于癌旁组织（P<0.001），且其蛋白表达水平与患者的组织学分级和淋巴结转移呈负相关（P<0.05）。Kaplan-Meier生存分析结果显示，DAZAP2低表达与乳腺癌患者不良预后相关（P<0.05）。CCK-8实验显示，与sh-Ctrl组相比，sh-DAZAP2-1组和sh-DAZAP2-2组乳腺癌细胞增殖活性升高（F=52.38,P<0.001）；与Flag组相比，FlagDAZAP2组细胞增殖活性降低（F=41.67,P<0.001）。克隆形成实验显示，与sh-Ctrl组相比，sh-DAZAP2-1组和sh-DAZAP2-2组克隆形成数增加（F=12.74,P<0.01）；与Flag组相比，Flag-DAZAP2组克隆形成数减少（t=9.86,P<0.001）。细胞划痕实验显示，与sh-Ctrl组相比，sh-DAZAP2-1组和sh-DAZAP2-2组划痕愈合率升高（F=18.23,P<0.01）；与Flag组相比，Flag-DAZAP2组划痕愈合率降低（t=10.42,P<0.001）。Transwell实验显示，与sh-Ctrl组相比，sh-DAZAP2-1组和sh-DAZAP2-2组细胞迁移及侵袭数量均增加（F=55.16、68.42，均P<0.001）；与Flag组相比，Flag-DAZAP2组细胞迁移及侵袭数量均减少（t=6.18、5.73，均P<0.01）。结论 DAZAP2高表达可通过抑制乳腺癌细胞的增殖、迁移及侵袭能力，延缓乳腺癌细胞的恶性进展。

Objective To investigate the expression of DAZ associated protein 2(DAZAP2) in breast cancer and its effect on the biological behavior of tumor cells.Methods The relationship between DAZAP2 expression and pa‑tient survival was analyzed using data from The Cancer Genome Atlas(TCGA) database. Immunohistochemistry(IHC) was performed to detect DAZAP2 expression in breast cancer tissues and paratumoral tissues, and its corre‑lation with clinicopathological characteristics was evaluated. Kaplan-Meier analysis was used to assess overall sur‑vival(OS) and recurrence-free survival(RFS) of patients with different DAZAP2 expression levels. Stable breast cancer cell lines with DAZAP2 knockdown or overexpression were established. The proliferation ability of breast cancer cells was evaluated by CCK-8 and colony formation assays, while migration and invasion abilities were de‑tected by wound healing and Transwell assays.Results Bioinformatics analysis and IHC results showed that DAZAP2 expression was significantly lower in breast cancer tissues than in paratumoral tissue(P< 0. 001). More‑over, DAZAP2 protein expression was negatively correlated with histological grade and lymph node metastasis(P<0. 05 for both). Kaplan-Meier survival analysis revealed that low DAZAP2 expression was associated with poor prognosis in breast cancer patients(P<0. 05). The CCK-8 assay showed that, compared with the sh-Ctrl group, the proliferative activity of breast cancer cells significantly increased in the sh-DAZAP2-1 and sh-DAZAP2-2 groups(F=52. 38, P<0. 001), whereas compared with the Flag group, cell proliferative activity significantly de‑creased in the Flag-DAZAP2 group(F=41. 67, P<0. 001). The colony formation assay demonstrated that, com‑pared with the sh-Ctrl group, the number of colonies significantly increased in the sh-DAZAP2-1 and sh-DAZAP2-2 groups(F=12. 74, P<0. 01), whereas compared with the Flag group, the number of colonies was significantly re‑duced in the Flag-DAZAP2 group(t=9. 86, P<0. 001). The wound-healing assay showed that, compared with the sh-Ctrl group, the wound closure rates significantly increased in the sh-DAZAP2-1 and sh-DAZAP2-2 groups(F=18. 23, P<0. 01), whereas compared with the Flag group, the wound closure rate significantly decreased in the Flag-DAZAP2 group(t=10. 42, P<0. 001). The Transwell assays showed that, compared with the sh-Ctrl group, the numbers of migrated and invaded cells significantly increased in the sh-DAZAP2-1 and sh-DAZAP2-2 groups(F=55. 16 and 68. 42, respectively; both P<0. 001), whereas compared with the Flag group, the numbers of mi‑grated and invaded cells significantly decreased in the Flag-DAZAP2 group(t=6. 18 and 5. 73, respectively; both P<0. 01).Conclusion High DAZAP2 expression can delay the malignant progression of breast cancer by sup‑pressing the proliferation, migration, and invasion of breast cancer cells.