Holliday junction recognition protein promotes pancreatic cancer growth and metastasis via modulation of the MDM2/p53 signaling
Holliday junction recognition protein promotes pancreatic cancer growth and metastasis via modulation of the MDM2/p53 signaling
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DOI：
10.1038/s41419-020-2595-9
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发表时间：
2020-05-21
影响因子：
9
通讯作者：
Cao, Yu
中科院分区：
文献类型：
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作者：
Wang, Chen-jing;Li, Xin;Cao, Yu
霍利迪连接体识别蛋白 (HJURP) 是一种组蛋白 H3 伴侣，与多种恶性肿瘤有关。然而，其在胰腺癌中的特征仍不清楚。通过 RT-qPCR、免疫印迹和免疫组织化学评估 PDAC 组织中 HJURP 的表达。使用 shRNA 或带有 HJURP 插入片段的质粒构建 HJURP 缺陷或过表达的 PDAC 细胞系。进行MTT、球形成实验、迁移和侵袭实验来评估PDAC细胞的活力、增殖、迁移和侵袭。我们使用异种移植小鼠模型来评估体内肿瘤的生长和转移。应用 RNA-seq 寻找 PDAC 中 HJURP 的潜在下游靶标，并通过包括免疫荧光在内的多种检测完成后续验证。此外，还进行了染色质免疫沉淀和荧光素酶报告基因测定，以探索 HJURP 通过 H3K4me2 对 MDM2 表达的潜在调节。在当前的研究中，我们发现PDAC细胞和组织中HJURP的表达显着高于邻近正常组织，并且高HJURP表达预示着较差的存活率。 HJURP在体外显着促进PDAC细胞的活力、球体形成、迁移和侵袭，在体内HJURP还促进肿瘤生长和转移。从机械角度来说，MDM2/p53 轴对于 HJURP 介导的 PDAC 恶性行为至关重要，并且 HJURP 通过 H3K4me2 调节 MDM2 表达。 HJURP 可以作为一种有前途的生物标志物，以及 PDAC 预后和治疗的目标。
Holliday junction recognition protein (HJURP) refers to a histone H3 chaperone that has been implicated in different kinds of malignancies. Yet, its character in pancreatic cancer remains unclear. The expression of HJURP was assessed in PDAC tissues by RT-qPCR, immunoblotting, and immunohistochemistry. HJURP-deficient or overexpressed PDAC cell lines were constructed, using shRNA or plasmids with HJURP insert. MTT, sphere formation assay, migration, and invasion assays were performed to evaluate the viability, proliferation, migration, and invasion of PDAC cells. We used xenograft mice models to assess the tumor growth and metastasis in vivo. RNA-seq was applicated in search of the potential downstream target of HJURP in PDAC and subsequent verification were fulfilled via multiple assays, including immunofluorescence. Additionally, chromatin immunoprecipitation and luciferase reporter assay were conducted to explore the potential regulation of MDM2 expression by HJURP through H3K4me2. In this current research, we found that the expression of HJURP in PDAC cells and tissue was significantly higher than those of adjacent normal tissue, and high HJURP expression predicted poor survival. HJURP significantly promoted the viability, sphere formation, migration, and invasion of PDAC cells in vitro, HJURP also facilitated tumor growth and metastasis in vivo. Mechanically, MDM2/p53 axis is critical for HJURP-mediated malignant behaviors in PDAC, and HJURP regulates MDM2 expression through H3K4me2. HJURP could serve as a promising biomarker, and target for PDAC prognosis and treatment.