Department of Molecular Medicine, Cell and Matrix Research Institute, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
BK21 Four KNU Convergence Educational Program of Biomedical Sciences for Creative Future Talents, Department of Biomedical Science, Kyungpook National University, Daegu, Republic of Korea.
Anticancer Res. 2023 May;43(5):1959-1965. doi: 10.21873/anticanres.16355.
BACKGROUND/AIM: Unique cartilage matrix-associated protein (UCMA), a recently discovered vitamin K-dependent protein (VKDP) with a large number of γ-carboxyglutamic acid (Gla) residues, is associated with ectopic calcifications. Although the function of VKDPs is related to their γ-carboxylation status, the carboxylation status of UCMA in breast cancer is still unknown. Here, we investigated the inhibitory effect of UCMA with differing γ-carboxylation status on breast cancer cell lines, such as MDA-MB-231, 4T1, and E0771 cells.
Undercarboxylated UCMA (ucUCMA) was generated by mutating the γ-glutamyl carboxylase (GGCX) recognition sites. The ucUCMA and carboxylated UCMA (cUCMA) proteins were collected from culture media of HEK293-FT cells that had been transfected with mutated GGCX and wild-type UCMA expression plasmids, respectively. Boyden Transwell and colony formation assays were performed to evaluate cancer cell migration, invasion, and proliferation.
Culture medium containing cUCMA protein inhibited the migration, invasion, and colony formation of MDA-MB-231 and 4T1 cells to a greater degree than medium containing ucUCMA protein. Significant reductions in the migration, invasion, and colony formation were also observed in cUCMA-treated E0771 cells compared to those in ucUCMA-treated cells.
The inhibitory role of UCMA in breast cancer is closely related to its γ-carboxylation status. The results of this study may be a basis for the development of UCMA-based anti-cancer drugs.
背景/目的：独特的软骨基质相关蛋白（UCMA）是一种最近发现的维生素 K 依赖性蛋白（VKDP），含有大量的γ-羧基谷氨酸（Gla）残基，与异位钙化有关。尽管 VKDP 的功能与其γ-羧化状态有关，但 UCMA 在乳腺癌中的羧化状态尚不清楚。在这里，我们研究了具有不同γ-羧化状态的 UCMA 对乳腺癌细胞系（如 MDA-MB-231、4T1 和 E0771 细胞）的抑制作用。
通过突变 γ-谷氨酰羧化酶（GGCX）识别位点生成非羧化 UCMA（ucUCMA）。ucUCMA 和羧化 UCMA（cUCMA）蛋白分别从转染突变 GGCX 和野生型 UCMA 表达质粒的 HEK293-FT 细胞的培养基中收集。Boyden Transwell 和集落形成实验用于评估癌细胞的迁移、侵袭和增殖。
含有 cUCMA 蛋白的培养基对 MDA-MB-231 和 4T1 细胞的迁移、侵袭和集落形成的抑制作用强于含有 ucUCMA 蛋白的培养基。在 cUCMA 处理的 E0771 细胞中，与 ucUCMA 处理的细胞相比，迁移、侵袭和集落形成也显著减少。
UCMA 对乳腺癌的抑制作用与其 γ-羧化状态密切相关。本研究结果可能为基于 UCMA 的抗癌药物的开发提供依据。
