Heng Win Sen, Pore Milind, Meijer Coby, Hiltermann T Jeroen N, Cheah Shiau-Chuen, Gosens Reinoud, Kruyt Frank A E
Department of Medical Oncology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
Department of Pulmonology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
Lung Cancer. 2021 Apr;154:13-22. doi: 10.1016/j.lungcan.2021.02.002. Epub 2021 Feb 12.
Cancer stem cells (CSCs) have been implicated in disease progression of aggressive cancers including small cell lung carcinoma (SCLC). Here, we have examined the possible contribution of CSCs to SCLC progression and aggressiveness.
GLC-14, GLC-16 and GLC-19 SCLC cell lines derived from one patient, representing increasing progressive stages of disease were used. CSC marker expressions was determined by RT-qPCR and western blotting analyses, and heterogeneity was studied by CSC marker expression by immunofluorescence microscopy and flow cytometry. Colony formation assays were used to assess stem cell properties and therapy sensitivity.
Increasing expression of stem cell markers MYC, SOX2 and particularly CD44 were found in association with advancing disease. Single and overlapping expression of these markers indicated the presence of different CSC populations. The accumulation of more homogeneous double- and triple-positive CSC populations evolved with disease progression. Functional characterization of CSC properties affirmed higher proficiency of colony forming ability and increased resistance to γ-irradiation in GLC-16 and GLC-19 compared to GLC-14. GLC-19 colony formation was significantly inhibited by a human anti-CD44 antibody.
The progressive increase of MYC, SOX2 and particularly CD44 expression that was accompanied with enhanced colony forming capacity and resistance in the in vitro GLC disease progression model, supports the potential clinical relevance of CSC populations in malignancy and disease relapse of SCLC.
癌症干细胞（CSCs）与侵袭性癌症（包括小细胞肺癌（SCLC））的疾病进展有关。在此，我们研究了CSCs对SCLC进展和侵袭性的可能作用。
使用了源自一名患者的GLC-14、GLC-16和GLC-19 SCLC细胞系，它们代表了疾病进展程度不断增加的阶段。通过RT-qPCR和蛋白质印迹分析确定CSC标志物表达，并通过免疫荧光显微镜和流式细胞术研究CSC标志物表达的异质性。采用集落形成试验评估干细胞特性和治疗敏感性。
发现干细胞标志物MYC、SOX2，尤其是CD44的表达增加与疾病进展相关。这些标志物的单一和重叠表达表明存在不同的CSC群体。随着疾病进展，更均匀的双阳性和三阳性CSC群体逐渐积累。与GLC-14相比，CSC特性的功能表征证实GLC-16和GLC-19具有更高的集落形成能力和对γ射线照射的抗性。人抗CD44抗体显著抑制GLC-19集落形成。
在体外GLC疾病进展模型中，MYC、SOX2，尤其是CD44表达的逐渐增加伴随着集落形成能力和抗性的增强，这支持了CSC群体在SCLC恶性肿瘤和疾病复发中的潜在临床相关性。
