Carboxypeptidase A4 negatively correlates with p53 expression and regulates the stemness of breast cancer cells.
Carboxypeptidase A4 negatively correlates with p53 expression and regulates the stemness of breast cancer cells.
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羧肽酶 A4 与 p53 表达负相关并调节乳腺癌细胞的干性
DOI：
10.7150/ijms.54954
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发表时间：
2021
影响因子：
3.6
通讯作者：
Sun L
中科院分区：
文献类型：
--
作者：
Wang Y;Xie Y;Niu Y;Song P;Liu Y;Burnett J;Yang Z;Sun D;Ran Y;Li Y;Sun L
背景：三阴性乳腺癌（TNBC）是一种侵袭性癌症亚型，缺乏有效的治疗选择，p53是最常发生突变或缺失的基因。羧肽酶A4（CPA4）是一种细胞外金属羧肽酶，与侵袭性密切相关。尽管最近一项研究表明CPA4可诱导乳腺癌细胞发生上皮 - 间质转化，但尚无研究探讨其与干细胞特性相关的功能以及CPA4与TNBC中p53的相关性。在本研究中，我们旨在研究乳腺癌组织中CPA4的水平，分析其与p53的关联，并探究其在癌症干细胞特性维持中的作用。
方法：分别使用在线数据库UALCAN（http://ualcan.path.uab.edu）和Kaplan - Meier绘图仪（www.kmplot.com）分析CPA4 mRNA水平及其预后价值。使用相应的一抗在商业组织芯片（上海生物芯片有限公司，中国上海）上通过免疫组化评估乳腺癌及相邻正常组织中CPA4、p53和ALDH1A1的表达。采用小干扰RNA敲低技术研究细胞增殖功能，并通过软琼脂集落形成实验和无血清培养基中的球体形成实验进行研究。
结果：对UALCAN数据集的分析发现，TNBC中CPA4 mRNA水平升高，尤其是在TP53突变亚组中。此外，乳腺癌患者中CPA4 mRNA高水平与不良的总生存期（OS）显著相关。免疫组化分析表明，32.1%的乳腺癌样本（45/140）中CPA4水平升高，乳腺癌组织中ALDH1A1和p53的阳性率分别为25%（35/140）和50%（70/140）。统计分析显示CPA4高水平与TNBC表型显著相关。相关性分析表明CPA4过表达与ALDH1A1呈正相关（P<0.01），与p53呈负相关（P<0.05）。在Kaplan - Meier生存分析中，CPA4或ALDH1A1高水平均与乳腺癌患者的不良生存显著相关。功能研究表明，CPA4的下调显著抑制了TNBC细胞增殖、软琼脂中的集落形成以及无血清培养基中的球体形成。
结论：本研究首次表明CPA4与p53表达呈负相关，并且抑制CPA4可减少具有干细胞特性的乳腺癌细胞数量。它可能是TNBC治疗的一个潜在靶点。
Background: Triple-negative breast cancer (TNBC) is an aggressive cancer subtype lacking effective treatment options, and p53 is the most frequently mutated or deleted gene. Carboxypeptidase A4 (CPA4) is an extracellular metallocarboxypeptidase, which was closely associated with aggressiveness. Although a recent study indicated that CPA4 could induce epithelial‑mesenchymal transition in breast cancer cells, no studies investigated its stemness-related function and the correlation between CPA4 and p53 in TNBC. In this study, we aimed to investigate the CPA4 levels in breast cancer tissues and analyze its association with p53, and study its roles in cancer stemness maintenance. Methods: CPA4 mRNA level and its prognostic value were analyzed by using online database UALCAN (http://ualcan.path.uab.edu) and Kaplan-Meier plotter (www.kmplot.com), respectively. The expression of CPA4, p53 and ALDH1A1 in breast cancer and adjacent normal tissues were evaluated by IHC using the corresponding primary antibodies on a commercial tissue array (Shanghai Biochip Co., Ltd., Shanghai, China). siRNA knockdown was used to study the function of proliferation, colony formation assay and sphere formation in serum-free medium. Results: Analysis of the UALCAN datasets identified that CPA4 mRNA levels were elevated in TNBC, especially in the TP53-mutant subgroup. Furthermore, high levels of CPA4 mRNA were significantly associated with unfavourable overall survival OS in breast cancer patients. Immunohistochemistical analysis demonstrated that CPA4 levels were elevated in 32.1% of breast cancer samples (45/140), and the positive rates of ALDH1A1 and p53 in the breast cancer tissues were 25% (35/140) and 50% (70/140), respectively. Statistical analysis revealed high levels of CPA4 was significantly associated with TNBC phenotype. Correlation analysis indicated that CPA4 over-expression was positively associated with ALDH1A1 (P<0.01) and negatively correlated with p53 (P<0.05). In Kaplan-Meier survival analysis, either high CPA4 or ALDH1A1 levels was significantly correlated with poor survival in breast cancer patients. Functional studies demonstrated that down-regulation of CPA4 significantly inhibited TNBC cell proliferation, colony-formation assays in soft agar and sphere formation in serum-free medium. Conclusion: This study demonstrated for the first time that CPA4 was negatively correlates with p53 expression and inhibition of CPA4 could reduce the number of breast cancer cells with stemness property. It might be a potential target for the TNBC treatment.
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影响因子：
4.6
作者：
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通讯作者：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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