Magan Mustafa, Wiechec Emilia, Roberg Karin
Division of Cell Biology, Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden.
Department of Otorhinolaryngology in Linköping, Anesthetics, Operations and Specialty Surgery Center, Region Östergötland, Linköping, Sweden.
Cancer Cell Int. 2020 Dec 22;20(1):599. doi: 10.1186/s12935-020-01718-6.
Head and neck squamous cell carcinoma (HNSCC) is a heterogeneous group of tumors for which the overall survival rate worldwide is around 60%. The tumor microenvironment, including cancer-associated fibroblasts (CAFs), is believed to affect the treatment response and migration of HNSCC. The aim of this study was to create a biologically relevant HNSCC in vitro model consisting of both tumor cells and CAFs cultured in 3D to establish predictive biomarkers for treatment response, as well as to investigate the impact of CAFs on phenotype, proliferation and treatment response in HNSCC cells.
Three different HNSCC patient-derived tumor cell lines were cultured with and without CAFs in a 3D model. Immunohistochemistry of the proliferation marker Ki67, epidermal growth factor receptor (EGFR) and fibronectin and a TUNEL-assay were performed to analyze the effect of CAFs on both tumor cell proliferation and response to cisplatin and cetuximab treatment in tumor spheroids (3D). mRNA expression of epithelial-mesenchymal transition (EMT) and cancer stem cells markers were analyzed using qRT-PCR.
The results demonstrated increased cell proliferation within the tumor spheroids in the presence of CAFs, correlating with increased expression of EGFR. In spheroids with increased expression of EGFR, a potentiated response to cetuximab treatment was observed. Surprisingly, an increase in Ki67 expressing tumor cells were observed in spheroids treated with cisplatin for 3 days, correlating with increased expression of EGFR. Furthermore, tumor cells co-cultured with CAFs presented an increased EMT phenotype compared to tumor cells cultured alone in 3D.
Taken together, our results reveal increased cell proliferation and elevated expression of EGFR in HNSCC tumor spheroids in the presence of CAFs. These results, together with the altered EMT phenotype, may influence the response to cetuximab or cisplatin treatment.
头颈部鳞状细胞癌（HNSCC）是一组异质性肿瘤，全球总体生存率约为60%。肿瘤微环境，包括癌症相关成纤维细胞（CAF），被认为会影响HNSCC的治疗反应和迁移。本研究的目的是创建一种生物学相关的体外HNSCC模型，该模型由在三维（3D）环境中培养的肿瘤细胞和CAF组成，以建立治疗反应的预测生物标志物，并研究CAF对HNSCC细胞表型、增殖和治疗反应的影响。
在3D模型中，对三种不同的源自HNSCC患者的肿瘤细胞系进行有无CAF的培养。进行增殖标志物Ki67、表皮生长因子受体（EGFR）和纤连蛋白的免疫组织化学以及TUNEL检测，以分析CAF对肿瘤球体（3D）中肿瘤细胞增殖以及对顺铂和西妥昔单抗治疗反应的影响。使用qRT-PCR分析上皮-间质转化（EMT）和癌症干细胞标志物的mRNA表达。
结果表明，在存在CAF的情况下，肿瘤球体内的细胞增殖增加，这与EGFR表达增加相关。在EGFR表达增加的球体中，观察到对西妥昔单抗治疗的增强反应。令人惊讶的是，在用顺铂处理3天的球体中，观察到Ki67表达的肿瘤细胞增加,这与EGFR表达增加相关。此外，与单独在3D中培养的肿瘤细胞相比，与CAF共培养的肿瘤细胞呈现出增加的EMT表型。
综上所述，我们的结果显示在存在CAF的情况下，HNSCC肿瘤球体中的细胞增殖增加且EGFR表达升高。这些结果，连同改变的EMT表型，可能会影响对西妥昔单抗或顺铂治疗的反应。
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