Department of Medicine I, Lichtenberg Research Group, Johannes Gutenberg University, Mainz, Germany.
Biotech Research and Innovation Centre (BRIC), Department of Health and Medical Science, University of Copenhagen, Copenhagen, Denmark.
Int J Cancer. 2019 Jun 1;144(11):2782-2794. doi: 10.1002/ijc.32026. Epub 2018 Dec 14.
Primary liver cancer (PLC) ranks among the most lethal solid cancers worldwide due to lack of effective biomarkers for early detection and limited treatment options in advanced stages. Development of primary culture models that closely recapitulate phenotypic and molecular diversities of PLC is urgently needed to improve the patient outcome. Long-term cultures of 7 primary liver cancer cell lines of hepatocellular and cholangiocellular origin were established using defined culture conditions. Morphological and histological characteristics of obtained cell lines and xenograft tumors were analyzed and compared to original tumors. Time course analyses of transcriptomic and genomic changes were performed using next-generation sequencing (NGS). Key oncogenic alterations were identified by targeted NGS and cell lines carrying potentially actionable mutations were treated with corresponding specific inhibitors. PDCL fully resembled morphological features of the primary cancers in vitro and in vivo over extended period in culture. Genomic alterations as well as transcriptome profiles showed high similarity with primary tumors and remained stable during long-term culturing. Targeted-NGS confirmed that key oncogenic mutations such as TP53, KRAS, CTNNB1 as well as actionable mutations (e.g. MET, cKIT, KDR) were highly conserved in PDCL and amenable for individualized therapeutic approaches. Integrative genomic and transcriptomic approaches further demonstrated that PDCL more closely resemble molecular and prognostic features of PLC than established cell lines and are valuable tool for direct target evaluation. Our integrative analysis demonstrates that PDCL represents refined model for discovery of relevant molecular subgroups and exploration of precision medicine approaches for the treatment of this deadly disease.
原发性肝癌 (PLC) 是全球最致命的实体癌症之一,主要原因是缺乏有效的早期检测生物标志物,以及晚期阶段治疗选择有限。迫切需要开发能够更紧密地重现 PLC 表型和分子多样性的原代培养模型,以改善患者的预后。使用明确的培养条件,建立了 7 株源自肝细胞和胆管细胞的原发性肝癌细胞系的长期原代培养。对获得的细胞系和异种移植肿瘤的形态学和组织学特征进行了分析,并与原始肿瘤进行了比较。使用下一代测序 (NGS) 对转录组和基因组的变化进行了时间过程分析。通过靶向 NGS 确定了关键的致癌改变,并且对携带潜在可操作突变的细胞系用相应的特异性抑制剂进行了治疗。PDCL 在体外和体内培养延长的时间内完全再现了原发性癌症的形态学特征。基因组改变以及转录组谱与原发性肿瘤具有高度相似性,并在长期培养过程中保持稳定。靶向 NGS 证实,TP53、KRAS、CTNNB1 等关键致癌突变以及可操作突变(如 MET、cKIT、KDR)在 PDCL 中高度保守,并且适用于个体化治疗方法。综合基因组和转录组方法进一步表明,PDCL 比已建立的细胞系更能模拟 PLC 的分子和预后特征,是直接评估靶标的有价值工具。我们的综合分析表明,PDCL 是发现相关分子亚群和探索治疗这种致命疾病的精准医学方法的精细模型。
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