Wen Xianhui, Cui Miaomiao, Zhang Junhua, Huang Hai
Center for Clinical Laboratories, The Affiliated Hospital of Guizhou Medical University, Guiyang, China.
School of Clinical Laboratory Science, Guizhou Medical University, Guiyang, China.
Front Immunol. 2025 Sep 1;16:1620390. doi: 10.3389/fimmu.2025.1620390. eCollection 2025.
Liquid-liquid phase separation (LLPS) plays an important role in the development of many tumors, including gastric cancer, but its prognostic value is unclear. The aim of this study was to explore the prognostic significance of LLPS-related genes in gastric cancer to provide a basis for improving the accuracy of prognostic prediction and finding potential therapeutic targets in gastric cancer.
Clinical and transcriptomic data of gastric cancer were downloaded from TCGA and GEO databases, and LLPS-related genes were extracted from PhaSepDB. Unsupervised clustering was used to identify molecular subtypes based on LLPS gene expression. LLPS gene features were constructed and validated by LASSO Cox regression, and their staging prediction value was also evaluated by machine learning methods. Key genes were validated by qRT-PCR, Western blot, immunofluorescence, and functional experiments (shRNA knockdown, CCK-8, clone formation, and scratch assay).
Twenty LLPS-associated genes showed significant mRNA expression, copy number variation, somatic mutation, and interaction network alterations in gastric cancer tissues. Two LLPS molecular isoforms with different survival outcomes and immune microenvironment characteristics were identified. A four-gene LLPS prognostic signature consisting of , and was constructed, and the high-risk group had a poorer prognosis and was prone to drug resistance. Machine learning analysis further confirmed the predictive value of this gene signature. Functional experiments showed that knockdown of PSPC1 significantly inhibited the proliferation (inhibition rate >50%, 0.001) and migration ability (<0.0001) of gastric cancer cells. Immunofluorescence confirmed the localization and aggregation characteristics of DACT1 and PSPC1.
This study revealed the important role of LLPS in gastric cancer, and the constructed four-gene LLPS signature is expected to be a novel biomarker for prognostic assessment and treatment of gastric cancer. PSPC1 plays a key role in gastric cancer progression, and has the value of a potential therapeutic target.
液-液相分离（LLPS）在包括胃癌在内的多种肿瘤发生发展中起重要作用，但其预后价值尚不清楚。本研究旨在探讨LLPS相关基因在胃癌中的预后意义，为提高预后预测准确性及寻找胃癌潜在治疗靶点提供依据。
从TCGA和GEO数据库下载胃癌临床及转录组数据，从PhaSepDB中提取LLPS相关基因。基于LLPS基因表达采用无监督聚类识别分子亚型。通过LASSO Cox回归构建并验证LLPS基因特征，同时采用机器学习方法评估其分期预测价值。通过qRT-PCR、蛋白质免疫印迹、免疫荧光及功能实验（shRNA敲低、CCK-8、克隆形成及划痕实验）验证关键基因。
20个LLPS相关基因在胃癌组织中显示出显著的mRNA表达、拷贝数变异、体细胞突变及相互作用网络改变。鉴定出两种具有不同生存结局和免疫微环境特征的LLPS分子亚型。构建了由[此处缺失基因名称]组成的四基因LLPS预后特征，高危组预后较差且易产生耐药性。机器学习分析进一步证实了该基因特征的预测价值。功能实验表明，敲低PSPC1可显著抑制胃癌细胞增殖（抑制率>50%，P<0.001）及迁移能力（P<0.0001）。免疫荧光证实了DACT1和PSPC1的定位及聚集特征。
本研究揭示了LLPS在胃癌中的重要作用，构建的四基因LLPS特征有望成为胃癌预后评估及治疗的新型生物标志物。PSPC1在胃癌进展中起关键作用，具有潜在治疗靶点价值。
