抗 PADI4 抗体通过抑制肿瘤微环境中的瓜氨酸化纤维连接蛋白来抑制乳腺癌。

Anti-PADI4 antibody suppresses breast cancer by repressing the citrullinated fibronectin in the tumor microenvironment.

机构信息

Medical Research Center of the Affiliated Hospital of Qingdao University, Wutaishan Road 1677, Qingdao, Shandong 266000, PR China.

Medical Research Center, the First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Jinan, Shandong 250014, PR China.

出版信息

Biomed Pharmacother. 2022 Sep;153:113289. doi: 10.1016/j.biopha.2022.113289. Epub 2022 Jun 27.

DOI:10.1016/j.biopha.2022.113289

PMID:35772376

Abstract

BACKGROUND

PADI4, an enzyme catalyzing arginine residues to citrulline residues, is highly expressed in malignant tumors. This study prepared a monoclonal anti-human PADI4 antibody and investigated the therapeutic effect of the antibody on breast tumors and the functional mechanism.

METHODS

After treatment with PADI4 antibody, the changes in tumor-bearing mice were examined by PET-CT, pathological assays, biochemical tests, routine blood tests, cytokine assays and metabolic assays. We used PADI4 recombinant protein to catalyze fibronectin (Fn) and then used citrullinated fibronectin (Cit-Fn) to culture MDA-MB-231 cells. We also treated Cit-Fn cultured cells with PADI4 antibody. The cultured cells were examined using cell proliferation, apoptosis, colony formation, migration and glycolic ATP production. Citrullination in the tumor tissues and peripheral blood was measured using Western blotting and ELISA, respectively.

RESULTS

Following PADI4 antibody treatment, tumor growth was significantly suppressed, and the number of apoptotic cells in tumor tissues was increased. The citrullination level in peripheral blood and tumor tissues was decreased, EMT-related gene expression in tumors was also decreased, and the spontaneous movement of tumor-bearing mice was increased following treatment. Following antibody treatment, the serum concentrations of IL-10, IL-12p70, IL-23, ALT and AST were significantly decreased. MDA-MB-231 cells treated with Cit-Fn showed increased cell proliferation, cell migration, colony formation and glycolytic ATP production and decreased apoptosis. The growth and migration of MDA-MB-231 cells were reduced following PADI4 antibody treatment, and PADI4 antibody inhibited the citrullination of fibronectin in vitro.

CONCLUSIONS

The PADI4 antibody had a therapeutic effect on breast tumors by inhibiting the citrullination of fibronectin to change the tumor tissue microenvironment. PADI4 antibody is a potential means for tumor treatment.

摘要

背景

PADI4 是一种能将精氨酸残基转化为瓜氨酸残基的酶，在恶性肿瘤中高度表达。本研究制备了一种单克隆抗人 PADI4 抗体，并研究了该抗体对乳腺癌的治疗作用及其功能机制。

方法

用 PADI4 抗体处理荷瘤小鼠后，通过 PET-CT、病理检测、生化试验、血常规检测、细胞因子检测和代谢试验观察肿瘤变化。用 PADI4 重组蛋白催化纤维连接蛋白（Fn），然后用瓜氨酸化纤维连接蛋白（Cit-Fn）培养 MDA-MB-231 细胞。我们还使用 PADI4 抗体处理 Cit-Fn 培养的细胞。用细胞增殖、凋亡、集落形成、迁移和糖酵解 ATP 产生试验检测培养的细胞。用 Western blot 和 ELISA 分别检测肿瘤组织和外周血中的瓜氨酸化水平。

结果

PADI4 抗体治疗后，肿瘤生长明显受到抑制，肿瘤组织中凋亡细胞增多。外周血和肿瘤组织中的瓜氨酸化水平降低，肿瘤中 EMT 相关基因表达降低，荷瘤小鼠的自发活动增加。抗体治疗后，血清中 IL-10、IL-12p70、IL-23、ALT 和 AST 浓度明显降低。用 Cit-Fn 处理的 MDA-MB-231 细胞表现出更高的细胞增殖、细胞迁移、集落形成和糖酵解 ATP 产生能力，以及更低的细胞凋亡率。PADI4 抗体治疗降低了 MDA-MB-231 细胞的生长和迁移能力，且体外抑制了纤维连接蛋白的瓜氨酸化。