National Center for Protein Sciences, the State Key Lab of Proteomics, Beijing Institute of Radiation Medicine, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 102206, China.
National Center for Protein Sciences, the State Key Lab of Proteomics, Beijing Institute of Life Omics, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 102206, China.
Yi Chuan. 2022 Mar 20;44(3):230-244. doi: 10.16288/j.yczz.21-426.
The abnormal expressions of minichromosome maintenance protein 2 (MCM2) are closely related to the development of various kinds of cancers. We aimed to explore the functions and potential molecular mechanisms of MCM2 gene in cholangiocarcinoma (CCA) cell lines (Huh28 and RBE). First, the cell counting kit-8 (CCK-8), plate clone formation, transwell and invasion assays showed that MCM2 promotes the proliferation, migration and invasion of CCA cells. Flow cytometry assays showed that MCM2 significantly promotes the cell cycle, and inhibits the apoptosis of CCA cells. Further, by analyzing the RNA sequencing data of cholangiocarcinoma, we found that MCM2 gene is significantly negatively correlated with p53 signaling pathway. Quantitative real time polymerase chain reaction (qRT-PCR) and Western blotting (WB) assays confirmed that MCM2 in CCA cells significantly down-regulated the mRNA and protein expression levels of p53 and BAX, and up-regulated the mRNA and protein expression levels of BCL2 and CCND1. Flow cytometry, qRT-PCR and WB assays confirmed that MCM2 promotes CCA through p53 pathway. Finally, we found that MCM2 is up-regulated in CCA tissues compared to the matched non-tumor cholangiocarcinoma tissues, and the high expressions of MCM2 are significantly associated with the poor clinical outcomes of CCA patients. In conclusion, this study revealed that MCM2 promotes the development of CCA by reducing the p53 pathway, and its high expression levels predict poor prognosis in CCA patients. These results provide a theoretical basis for the development of new clinical diagnosis and treatment of cholangiocarcinoma in the future.
微小染色体维持蛋白 2（MCM2）的异常表达与各种癌症的发展密切相关。我们旨在探讨 MCM2 基因在胆管癌细胞系（Huh28 和 RBE）中的功能和潜在分子机制。首先，细胞计数试剂盒-8（CCK-8）、平板克隆形成、Transwell 和侵袭实验表明，MCM2 促进 CCA 细胞的增殖、迁移和侵袭。流式细胞术实验表明，MCM2 显著促进细胞周期，并抑制 CCA 细胞的凋亡。进一步，通过分析胆管癌的 RNA 测序数据，我们发现 MCM2 基因与 p53 信号通路呈显著负相关。定量实时聚合酶链反应（qRT-PCR）和 Western blot（WB）实验证实，MCM2 在 CCA 细胞中显著下调 p53 和 BAX 的 mRNA 和蛋白表达水平，而上调 BCL2 和 CCND1 的 mRNA 和蛋白表达水平。流式细胞术、qRT-PCR 和 WB 实验证实，MCM2 通过 p53 通路促进 CCA。最后，我们发现与配对的非肿瘤性胆管癌组织相比，MCM2 在 CCA 组织中上调，并且 MCM2 的高表达与 CCA 患者的不良临床结局显著相关。总之，本研究揭示了 MCM2 通过降低 p53 通路促进 CCA 的发展，其高表达水平预示着 CCA 患者预后不良。这些结果为未来胆管癌的新临床诊断和治疗提供了理论依据。
