Su B B, Zhang C, Wei B S, Cao J, Peng R, Tu D Y, Jiang G Q, Jin S J, Bai D S
Department of Hepatobiliary Surgery, Northern Jiangsu People's Hospital Affiliated to Yangzhou University, Yangzhou 225001, China.
Department of Hepatobiliary Surgery, Northern Jiangsu People's Hospital Affiliated to Yangzhou University, Yangzhou 225001, China Department of Hepatobiliary Surgery, Northern Jiangsu People's Hospital, Yangzhou 225001, China.
Zhonghua Gan Zang Bing Za Zhi. 2025 Mar 20;33(3):237-247. doi: 10.3760/cma.j.cn501113-20240827-00396.
To investigate the expression, prognosis, and role of G protein-coupled receptor kinase-interacting protein 1 (GIT1) in patients with hepatocellular carcinoma (HCC) tumor micro environments. Clinical data of 140 cases who underwent complete HCC surgical resection from January 2015 to December 2021 in Subei People's Hospital affiliated to Yangzhou University, Jiangsu Province, were included. Tumor tissue and adjacent tissue samples were collected for immunohistochemical analysis. The patients were divided into a high expression group and a low expression group according to the expression of GIT1. Cox regression was used to analyze the risk factors for prognosis in patients with HCC. Fifteen pairs of cancer tissues and adjacent tissues were randomly matched for quantitative polymerase chain reaction (RT-PCR), western blot (WB), and immunohistochemical analysis. GITI knockout or overexpression cell lines of human hepatoma cell lines HepG2, HuH7 and MHCC97-H, and mouse hepatoma cell line Hepa 1-6 were constructed. The effects on M2 macrophage polarization were analyzed by flow cytometry. A mice tumor model was constructed. The growth curve of tumor tissue overexpressing GIT1 was plotted. Bioinformatics analysis of the Cancer Genome Atlas (TCGA) data was performed using OncoLnc, Kaplan-Meier Plotter, UALCAN, and GEPIA databases to explore the differential expression of GIT1 in HCC patients and its effect on prognosis. Bioinformatics analysis showed that the expression level of GIT1 was significantly higher in HCC tissues than in normal liver tissues (<0.05). RT-PCR and WB experiments showed that GIT1 was highly expressed in HCC. The follow-up results showed that high expression of GIT1 was associated with poor prognosis in patients with HCC. The high expression of GIT1 was an independent risk factor for the prognosis in patients with HCC (=2.562, 95%: 0.231-0.704, <0.05). Functional enrichment analysis combined with TIMER database analysis found that GIT1 expression level was associated with multiple immune cell infiltrations in HCC, but the correlation coefficient with macrophage infiltration was the highest (=0.545, <0.001). Mice tumorigenesis experiments showed that the tumor volume of GIT1-overexpressing mice was significantly increased (<0.05). Additionally, flow cytometry indicated that after GIT1 overexpression, there was a low degree of M1 infiltration/polarization (wild type: 5.06%±0.11%, overexpression type: 4.09%±0.04%; <0.05) and a high degree of M2 infiltration/polarization (wild type: 10.20%±0.33%, overexpression type: 14.7%±0.12%; <0.05). GIT1 serves as a prognostic biomarker in HCC, promoting tumor progression through its high expression and enhances M2 macrophage infiltration.
探讨G蛋白偶联受体激酶相互作用蛋白1（GIT1）在肝细胞癌（HCC）肿瘤微环境中的表达、预后及作用。纳入2015年1月至2021年12月在江苏省扬州大学附属苏北人民医院接受HCC完整手术切除的140例患者的临床资料。收集肿瘤组织和相邻组织样本进行免疫组织化学分析。根据GIT1的表达将患者分为高表达组和低表达组。采用Cox回归分析HCC患者预后的危险因素。随机匹配15对癌组织和相邻组织进行定量聚合酶链反应（RT-PCR）、蛋白质免疫印迹（WB）和免疫组织化学分析。构建人肝癌细胞系HepG2、HuH7和MHCC97-H以及小鼠肝癌细胞系Hepa 1-6的GIT1基因敲除或过表达细胞系。通过流式细胞术分析对M2巨噬细胞极化的影响。构建小鼠肿瘤模型。绘制过表达GIT1的肿瘤组织生长曲线。使用OncoLnc、Kaplan-Meier Plotter、UALCAN和GEPIA数据库对癌症基因组图谱（TCGA）数据进行生物信息学分析，以探索GIT1在HCC患者中的差异表达及其对预后的影响。生物信息学分析显示，GIT1在HCC组织中的表达水平显著高于正常肝组织（P<0.05）。RT-PCR和WB实验表明GIT1在HCC中高表达。随访结果显示，GIT1高表达与HCC患者预后不良相关。GIT1高表达是HCC患者预后的独立危险因素（P=2.562，95%CI：0.231-0.704，P<0.05）。功能富集分析结合TIMER数据库分析发现，GIT1表达水平与HCC中多种免疫细胞浸润相关，但与巨噬细胞浸润的相关系数最高（r=0.545，P<0.001）。小鼠肿瘤发生实验表明，过表达GIT1的小鼠肿瘤体积显著增加（P<0.05）。此外，流式细胞术表明，GIT1过表达后，M1浸润/极化程度较低（野生型：5.06%±0.11%，过表达型：4.09%±0.04%；P<0.05），M2浸润/极化程度较高（野生型：10.20%±0.33%，过表达型：14.7%±0.12%；P<0.05）。GIT1作为HCC的预后生物标志物，通过其高表达促进肿瘤进展并增强M2巨噬细胞浸润。
