Blandino Alice, Scherer Dominique, Boekstegers Felix, Rounge Trine B, Langseth Hilde, Roessler Stephanie, Hveem Kristian, Brenner Hermann, Pechlivanis Sonali, Waldenberger Melanie, Lorenzo Bermejo Justo
Statistical Genetics Research Group, Institute of Medical Biometry, Heidelberg University, Heidelberg, Germany.
Department of Research, Cancer Registry of Norway, Norwegian Institute of Public Health, Oslo, Norway; Center for Bioinformatics, Department of Pharmacy, University of Oslo, Oslo, Norway.
Eur J Cancer. 2025 Jan;214:115138. doi: 10.1016/j.ejca.2024.115138. Epub 2024 Nov 20.
Gallbladder cancer (GBC) is an aggressive disease with limited treatment options but high prevention potential. GBC tumours take 10-20 years to develop, a timeframe that holds potential for early detection. MicroRNAs (miRNAs) play a central role in abnormal cell processes, and circulating miRNAs may constitute valuable biomarkers of early disease. We used microarray data to pre-select differentially expressed miRNAs in formalin-fixed paraffin-embedded (FFPE) gallbladder tissue samples (GBC n = 40, normal n = 8). We then applied small-RNA sequencing to screen for miRNA expression differences in serum samples from three European prospective cohorts (n = 37 GBC case-control pairs), and validated the most promising candidates in three independent cohorts (n = 36 GBC case- control pairs). Statistical analyses included robust linear regression, pathway and meta-analysis, and examination of expression correlation between miRNAs and target genes. MiR-4533 and miR-671-5p were overexpressed in GBC tissue and serum samples, and meta-analysis confirmed the overexpression of miR-4533 in GBC serum samples from the prospective cohorts (p-value = 4.1×10), especially in individuals of female sex, under 63.5 years, or with a BMI below 26.2 kg/m. Pathway and correlation analyses revealed that miR-4533 targets SIPA1L2 in the Rap1 signalling pathway, and SIPA1L2 was downregulated in GBC serum samples. Our study highlights the advantage of integrating small-RNA sequencing results from different types of samples and independent datasets, and the need for international research collaborations to identify and validate biomarkers for secondary prevention of rare tumours such as GBC. The function of miR-4533 and its interaction with SIPA1L2 in GBC development need to be further investigated.
胆囊癌（GBC）是一种侵袭性疾病，治疗选择有限，但具有较高的预防潜力。GBC肿瘤需要10至20年的时间发展，这一时间框架为早期检测提供了可能性。微小RNA（miRNA）在异常细胞过程中起核心作用，循环miRNA可能构成早期疾病的有价值生物标志物。我们使用微阵列数据在福尔马林固定石蜡包埋（FFPE）胆囊组织样本中预先选择差异表达的miRNA（GBC组n = 40，正常组n = 8）。然后，我们应用小RNA测序来筛选来自三个欧洲前瞻性队列的血清样本中的miRNA表达差异（n = 37对GBC病例对照），并在三个独立队列（n = 36对GBC病例对照）中验证最有前景的候选物。统计分析包括稳健线性回归、通路和荟萃分析，以及检查miRNA与靶基因之间的表达相关性。MiR-4533和miR-671-5p在GBC组织和血清样本中过表达，荟萃分析证实了miR-4533在前瞻性队列的GBC血清样本中过表达（p值 = 4.1×10），特别是在63.5岁以下、BMI低于26.2 kg/m的女性个体中。通路和相关性分析表明，miR-4533靶向Rap1信号通路中的SIPA1L2，并且SIPA1L2在GBC血清样本中下调。我们的研究强调了整合来自不同类型样本和独立数据集的小RNA测序结果的优势，以及开展国际研究合作以识别和验证罕见肿瘤（如GBC）二级预防生物标志物的必要性。miR-4533的功能及其在GBC发展中与SIPA1L2的相互作用需要进一步研究。
