Abstract：
OBJECTIVE: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
摘要：
目标：独特软骨基质相关蛋白（UCMA），最近发现的维生素K依赖性蛋白（VKDP）具有大量的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，乳腺癌中UCMA的羧化状态尚不清楚.这里,我们研究了不同γ-羧化状态的UCMA对乳腺癌细胞系的抑制作用，例如MDA-MB-231、4T1和E0771细胞。
方法：通过突变γ-谷氨酰羧化酶（GGCX）识别位点产生羧化不足的UCMA（ucUCMA）。从已经用突变GGCX和野生型UCMA表达质粒转染的HEK293-FT细胞的培养基中收集ucUCMA和羧化UCMA（cUCMA）蛋白，分别。进行BoydenTranswell和集落形成测定以评估癌细胞迁移，入侵,和扩散。
结果：含有cUCMA蛋白的培养基抑制迁移，入侵,MDA-MB-231和4T1细胞的集落形成程度高于含ucUCMA蛋白的培养基。移民大幅减少，入侵,与ucUCMA处理的细胞相比,在cUCMA处理的E0771细胞中也观察到集落形成。
结论：UCMA对乳腺癌的抑制作用与其γ-羧化状态密切相关。这项研究的结果可能是开发基于UCMA的抗癌药物的基础。
方法：通过突变γ-谷氨酰羧化酶（GGCX）识别位点产生羧化不足的UCMA（ucUCMA）。从已经用突变GGCX和野生型UCMA表达质粒转染的HEK293-FT细胞的培养基中收集ucUCMA和羧化UCMA（cUCMA）蛋白，分别。进行BoydenTranswell和集落形成测定以评估癌细胞迁移，入侵,和扩散。
结果：含有cUCMA蛋白的培养基抑制迁移，入侵,MDA-MB-231和4T1细胞的集落形成程度高于含ucUCMA蛋白的培养基。移民大幅减少，入侵,与ucUCMA处理的细胞相比,在cUCMA处理的E0771细胞中也观察到集落形成。
结论：UCMA对乳腺癌的抑制作用与其γ-羧化状态密切相关。这项研究的结果可能是开发基于UCMA的抗癌药物的基础。
