DNA 引发酶亚基 1 通过激活 AKT/mTOR 信号通路和 UBE2C 介导的 P53 泛素化作用，恶化了肝细胞癌的进展。
DNA primase subunit 1 deteriorated progression of hepatocellular carcinoma by activating AKT/mTOR signaling and UBE2C-mediated P53 ubiquitination.
作者信息
Zhu Mengqi, Wu Mengna, Bian Saiyan, Song Qianqian, Xiao Mingbing, Huang Hua, You Li, Zhang Jianping, Zhang Jie, Cheng Chun, Ni Wenkai, Zheng Wenjie
机构信息
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, 20 Xisi Road, Nantong, 226001, Jiangsu, China.
School of Medicine, Nantong University, 19 Qixiu Road, Nantong, 226001, Jiangsu, China.
出版信息
Cell Biosci. 2021 Feb 23;11(1):42. doi: 10.1186/s13578-021-00555-y.
BACKGROUND
DNA primase subunit 1 (PRIM1) has been reported as a novel oncogene in several cancer types. However, its roles in hepatocellular carcinoma (HCC) remain unclear. This study aimed to investigate underlying mechanisms of PRIM1 and identify it as a potential molecular target for HCC.
METHODS
Hub genes were screened between HCC tissues and normal liver tissues in 3 gene expression omnibus (GEO) datasets and the cancer genome atlas (TCGA). The expression features and prognostic value of one of the hub genes PRIM1 were analyzed by bioinformatic analyses and immunohistochemistry. Loss-of-function and gain-of-function studies were used to investigate the regulatory role of PRIM1 in HCC cells. Real-time (RT)-qPCR, western blotting, and ubiquitin immunoprecipitation assays were performed to explore the underlying mechanisms. The xenograft model was employed to detect the roles of PRIM1 in tumor growth in vivo. Finally, the 3D spheroid model was conducted to validate the role of PRIM1 in tumor growth and sorafenib resistance.
RESULTS
The hub genes of HCC were screened in multiple bioinformatic datasets. PRIM1, as one of the hub genes, was significantly overexpressed in HCC tissues in mRNA and protein levels. In addition, high expression of PRIM1 indicated poor prognosis of HCC patients in TCGA, ICGC, and Nantong cohorts. Overexpression of PRIM1 promoted the proliferation, migration/invasion, and sorafenib resistance of HCC cells, with the decrease in apoptosis and cell cycle arrest. Mechanically, PRIM1 facilitated epithelial-mesenchymal transition (EMT) process and the activity of PI3K/AKT/mTOR signaling of HCC cells. Additionally, PRIM1 could cause the ubiquitination and degradation of P53 by upregulating Ubiquitin Conjugating Enzyme E2 C (UBE2C). Furthermore, knockdown of PRIM1 significantly inhibited the growth of xenograft tumors and HCC cells-derived spheroids with enhanced sorafenib resistance.
CONCLUSION
This study implies that PRIM1 may play a key role in the progression of HCC and may serve as a potential target for HCC treatment.
背景
DNA 引发酶亚基 1（PRIM1）在多种癌症类型中已被报道为一种新型致癌基因。然而，其在肝细胞癌（HCC）中的作用仍不清楚。本研究旨在探讨 PRIM1 的潜在机制，并将其鉴定为 HCC 的潜在分子靶点。
方法
在 3 个基因表达综合数据库（GEO）数据集和癌症基因组图谱（TCGA）中筛选 HCC 组织和正常肝组织之间的枢纽基因。通过生物信息学分析和免疫组织化学分析枢纽基因 PRIM1 之一的表达特征和预后价值。采用功能丧失和功能获得研究来探讨 PRIM1 在 HCC 细胞中的调节作用。进行实时（RT）-qPCR、蛋白质印迹和泛素免疫沉淀试验以探索潜在机制。采用异种移植模型检测 PRIM1 在体内肿瘤生长中的作用。最后，进行三维球体模型以验证 PRIM1 在肿瘤生长和索拉非尼耐药性中的作用。
结果
在多个生物信息学数据集中筛选出 HCC 的枢纽基因。PRIM1 作为枢纽基因之一，在 HCC 组织中的 mRNA 和蛋白质水平均显著过表达。此外，PRIM1 的高表达表明在 TCGA、ICGC 和南通队列中 HCC 患者的预后较差。PRIM1 的过表达促进了 HCC 细胞的增殖、迁移/侵袭和索拉非尼耐药性，同时细胞凋亡减少和细胞周期停滞。机制上，PRIM1 促进了 HCC 细胞的上皮-间质转化（EMT）过程和 PI3K/AKT/mTOR 信号通路的活性。此外，PRIM1 可通过上调泛素结合酶 E2 C（UBE2C）导致 P53 的泛素化和降解。此外，敲低 PRIM1 显著抑制了异种移植肿瘤和 HCC 细胞来源球体的生长，并增强了索拉非尼耐药性。
结论
本研究表明 PRIM1 可能在 HCC 的进展中起关键作用，并可能作为 HCC 治疗的潜在靶点。
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