从淋巴结中扩增体外培养的肺癌来源的浸润癌细胞，鉴定与转移进展相关的细胞。
Ex vivo expansion of lung cancer-derived disseminated cancer cells from lymph nodes identifies cells associated with metastatic progression.
发表日期：2023 Aug 09
作者：
Steffi Treitschke, Kathrin Weidele, Adithi Ravikumar Varadarajan, Giancarlo Feliciello, Jens Warfsmann, Sybille Vorbeck, Bernhard Polzer, Catherine Botteron, Martin Hoffmann, Vadim Dechand, Tobias Mederer, Florian Weber, Melanie Werner-Klein, Tobias Robold, Hans-Stefan Hofmann, Christian Werno, Christoph A Klein
来源：
INTERNATIONAL JOURNAL OF CANCER
摘要：
肺癌的明显侵袭性的细胞基础尚不清楚，但很可能与转移性癌细胞（DCCs）的功能或分子特征有关。上皮癌细胞的DCCs主要通过针对细胞角蛋白或EpCAM等组织发生标志物的抗体检测得到。有人认为，标志物阴性的转移性创始细胞可能会逃避检测。因此，我们利用体外球体形成进行候选转移创始者的功能检测。我们从131例肺癌患者的199个淋巴结样本中制备细胞悬液，并将其放入非附着的细胞培养中。球体形成与使用EpCAM免疫细胞学检测DCCs以及显著较差的预后相关。球体形成的预后影响与EpCAM阳性DCCs的数量较多和扩展球体的异常基因型密切相关。我们还注意到，在没有淋巴扩散证据的患者中也有球体形成，然而这些球体的特征基因（与EpCAM阳性样本的球体相关）表达不频繁，并且既没有典型的肺癌突变（KRAS，TP53，ERBB1），也没有拷贝数变异，但可能与在根治手术后的5年以上的疾病进展有关。我们得出结论，EpCAM可以识别与疾病进展相关的DCCs，并且可以扩展此类细胞用于模型生成，还需进一步研究以阐明罕见的EpCAM阴性球体形成细胞的功能和预后作用。© 2023 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.
The cellular basis of the apparent aggressiveness in lung cancer is poorly understood but likely associated with functional or molecular features of disseminated cancer cells (DCCs). DCCs from epithelial cancers are mostly detected by antibodies directed against histogenetic markers such as cytokeratin or EpCAM. It has been argued that marker-negative metastatic founder cells might escape detection. We therefore used ex vivo sphere formation for functional detection of candidate metastasis founders. We generated cell suspensions from 199 LN samples of 131 lung cancer patients and placed them into non-adherent cell culture. Sphere formation was associated with detection of DCCs using EpCAM immunocytology and with significantly poorer prognosis. The prognostic impact of sphere formation was strongly associated with high numbers of EpCAM-positive DCCs and aberrant genotypes of expanded spheres. We also noted sphere formation in patients with no evidence of lymphatic spread, however such spheres showed infrequent expression of signature genes associated with spheres from EpCAM-positive samples and displayed neither typical lung cancer mutations (KRAS, TP53, ERBB1) nor copy number variations, but might be linked to disease progression >5 years post curative surgery. We conclude that EpCAM identifies relevant disease-driving DCCs, that such cells can be expanded for model generation and that further research is needed to clarify the functional and prognostic role of rare EpCAM-negative sphere forming cells.© 2023 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.