Datasheet1_Classification of the mitochondrial ribosomal protein-associated molecular subtypes and identified a serological diagnostic biomarker in hepatocellular carcinoma.zip

PurposeThe objective of this study was to sort out innovative molecular subtypes associated with mitochondrial ribosomal proteins (MRPs) to predict clinical therapy response and determine the presence of circulating markers in hepatocellular carcinoma (HCC) patients.MethodsUsing an unsupervised clustering method, we categorized the relative molecular subtypes of MRPs in HCC patients. The prognosis, biological properties, immune checkpoint inhibitor and chemotherapy response of the patients were clarified. A signature and nomogram were developed to evaluate the prognosis. Enzyme-linked immunosorbent assay (ELISA) measured serum mitochondrial ribosomal protein L9 (MRPL9) levels in liver disease patients and normal individuals. Receiver operating characteristic (ROC) curves were conducted to calculate the diagnostic effect. The Cell Counting Kit 8 was carried out to examine cell proliferation, and flow cytometry was used to investigate the cell cycle. Transwell assay was applied to investigate the potential of cell migration and invasion. Western blot detected corresponding changes of biological markers.ResultsParticipants were classified into two subtypes according to MRPs expression levels, which were characterized by different prognoses, biological features, and marked differences in response to chemotherapy and immune checkpoint inhibitors. Serum MRPL9 was significantly higher in HCC patients than in normal individuals and the benign liver disease group. ROC curve analysis showed that MRPL9 was superior to AFP and Ferritin in differentiating HCC from healthy and benign patients, or alone. Overexpressed MRPL9 could enhance aggressiveness and facilitate the G1/S progression in HCC cells.ConclusionWe constructed novel molecular subtypes based on MRPs expression in HCC patients, which provided valuable strategies for the prediction of prognosis and clinical personalized treatment. MRPL9 might act as a reliable circulating diagnostic biomarker and therapeutic target for HCC patients.

本研究旨在梳理与线粒体核糖体蛋白（MRPs）相关的创新分子亚型，以预测临床治疗效果并确定肝细胞癌（HCC）患者体内循环标志物的存在。研究方法包括采用无监督聚类方法对HCC患者中MRPs的相对分子亚型进行分类，明确患者的预后、生物学特性、免疫检查点抑制剂和化疗反应。本研究开发了一套标志物和列线图以评估预后。通过酶联免疫吸附测定（ELISA）检测肝病患者及健康个体血清中线粒体核糖体蛋白L9（MRPL9）水平，并采用受试者工作特征（ROC）曲线计算其诊断效果。细胞计数试剂盒8用于检测细胞增殖，流式细胞术用于研究细胞周期。Transwell实验用于探究细胞迁移和侵袭的潜力。蛋白质印迹法检测生物标志物的相应变化。根据MRPs表达水平，参与者被分为两种亚型，这些亚型以不同的预后、生物学特征以及显著的化疗和免疫检查点抑制剂反应差异为特征。与正常个体和良性肝病组相比，HCC患者的血清MRPL9水平显著升高。ROC曲线分析表明，MRPL9在区分HCC与健康和良性患者方面优于甲胎蛋白（AFP）和铁蛋白。过表达的MRPL9可能增强HCC细胞的侵袭性并促进G1/S期的进展。结论：基于HCC患者MRPs表达构建的新型分子亚型为预后预测和临床个性化治疗提供了宝贵的策略。MRPL9可能作为HCC患者可靠的循环诊断生物标志物和治疗靶点。