Clinical Laboratory Center, Cancer Hospital Affiliated to Xinjiang Medical University, Xinjiang, China.
J Transl Med. 2024 Jan 2;22(1):11. doi: 10.1186/s12967-023-04837-6.
Guanylate binding protein 1 (GBP1) is the most concerned member of the GBP family, which has a series of effects such as anti-infection and anti-angiogenesis. Its role in malignant tumors including cervical cancer is still controversial. We aim to explore the effects of GBP1 on cervical cancer through bioinformatics and related experiments. In this study, we first found that GBP1 was generally expressed in cervical cancer in various online databases and was closely related to immune invasion. Secondly, we used multicolor immunofluorescence technology to verify the expression of GBP1 in cervical cancer tissues and its relationship with immune invasion, and explored its relationship with the prognosis of patients with cervical cancer. Knockdown and overexpression assays of GBP1 in vitro were used to prove GBP1 as a potential oncogene of cervical cancer, and its carcinogenicity was verified by in vivo experiment. In order to explore the potential mechanism of GBP1 in promoting cancer, RNA-seq was performed on GBP1 overexpression and knockdown expression cell lines, and GBP1 knockdown and overexpression were found to be associated with many RNA alternative splicing events, suggesting that GBP1 maybe a RNA binding protein (RBP) which affect the biological characteristics of cervical cancer cells through the alternative splicing pathway. However, the later RNA binding protein immunoprecipitation (RIP) assay proved that GBP1 was not a direct alternative splicing factor, while the co-immunoprecipitation (CoIP)-mass spectroscopy (MS) assay combined with protein protein interaction (PPI) analysis proved that 8 alternative splicing factors including Heterogeneous Nuclear Ribonucleoprotein K (HNRNPK) were interacting proteins of GBP1. Combined with the existing reports and the results of RNA-seq alternative splicing analysis, it is speculated that GBP1 may regulate the alternative splicing of CD44 protein by binding to interacting protein-HNRNPK, and thus play a role in promoting cancer in cervical cancer.
鸟苷酸结合蛋白 1（GBP1）是 GBP 家族中最受关注的成员，具有抗感染和抗血管生成等一系列作用。其在宫颈癌等恶性肿瘤中的作用仍存在争议。我们旨在通过生物信息学和相关实验探讨 GBP1 对宫颈癌的影响。在本研究中，我们首先在多个在线数据库中发现 GBP1 在宫颈癌中普遍表达，并与免疫浸润密切相关。其次，我们使用多色免疫荧光技术验证了 GBP1 在宫颈癌组织中的表达及其与免疫浸润的关系，并探讨了其与宫颈癌患者预后的关系。体外 GBP1 敲低和过表达实验用于证明 GBP1 是宫颈癌的潜在癌基因，并通过体内实验验证了其致癌性。为了探讨 GBP1 促进癌症的潜在机制，我们对 GBP1 过表达和敲低表达细胞系进行了 RNA-seq 分析，发现 GBP1 敲低和过表达与许多 RNA 可变剪接事件相关，提示 GBP1 可能是一种 RNA 结合蛋白（RBP），通过可变剪接途径影响宫颈癌细胞的生物学特性。然而，随后的 RNA 结合蛋白免疫沉淀（RIP）实验证明 GBP1 不是直接的可变剪接因子，而共免疫沉淀（CoIP）-质谱（MS）实验结合蛋白质相互作用（PPI）分析证明 8 种可变剪接因子包括异质核核糖核蛋白 K（HNRNPK）是 GBP1 的相互作用蛋白。结合现有报道和 RNA-seq 可变剪接分析结果，推测 GBP1 可能通过与相互作用蛋白 HNRNPK 结合来调节 CD44 蛋白的可变剪接，从而在宫颈癌中发挥促癌作用。
