Spheroid Culture of Head and Neck Cancer Cells Reveals an Important Role of EGFR Signalling in Anchorage Independent Survival.
Spheroid Culture of Head and Neck Cancer Cells Reveals an Important Role of EGFR Signalling in Anchorage Independent Survival.
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DOI：
10.1371/journal.pone.0163149
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Tinhofer I
中科院分区：
文献类型：
--
作者：
Braunholz D;Saki M;Niehr F;Öztürk M;Borràs Puértolas B;Konschak R;Budach V;Tinhofer I
在实体瘤中，每天有数百万个细胞流入血液循环。这些循环肿瘤细胞（CTC）中只有一部分存活，其中许多可能是因为它们形成多细胞簇（也称为球体）的潜力。这些球状体内的肿瘤细胞受到保护，免受失巢凋亡，这使得它们转移到远处器官或在原发部位重新接种。我们使用头颈部鳞状细胞癌（HNSCC）细胞系的球状体培养物作为此类CTC簇的模型，以确定表皮生长因子受体（EGFR）在簇形成能力和从细胞外基质脱离后细胞存活中的作用。HNSCC细胞系FaDu、SCC-9和UT-SCC-9（UT-SCC-9 P）及其西妥昔单抗（CTX）抗性亚克隆（UT-SCC-9 R）通过用抗粘附聚合物聚甲基丙烯酸2-羟乙酯（poly-HEMA）包被培养皿而被迫以锚定非依赖性方式生长。在这种培养条件下的细胞凋亡，克隆存活和EGFR信号转导的程度进行了评估。发现悬浮培养中球体形成的潜力与HNSCC细胞系的增殖率以及它们的基础EGFR表达水平正相关。CTX和吉非替尼阻断，而EGFR配体的加入促进了锚定非依赖性细胞存活和球体形成。球状体形成和生长的增加与EGFR及其下游信号成分（MAPK/ERK）的持续活化相关。重要的是，来源于球状体培养物的HNSCC细胞在不存在细胞-基质接触的情况下保留其克隆形成潜力。在这些条件下加入CTX强烈抑制CTX敏感细胞系的集落形成，但不抑制其耐药亚克隆。总之，EGFR活化被确定为HNSCC细胞的锚定非依赖性存活的关键因素。在CTC簇形成中靶向EGFR可能代表HNSCC中有吸引力的抗转移治疗方法。
In solid tumours millions of cells are shed into the blood circulation each day. Only a subset of these circulating tumour cells (CTCs) survive, many of them presumable because of their potential to form multi-cellular clusters also named spheroids. Tumour cells within these spheroids are protected from anoikis, which allows them to metastasize to distant organs or re-seed at the primary site. We used spheroid cultures of head and neck squamous cell carcinoma (HNSCC) cell lines as a model for such CTC clusters for determining the role of the epidermal growth factor receptor (EGFR) in cluster formation ability and cell survival after detachment from the extra-cellular matrix. The HNSCC cell lines FaDu, SCC-9 and UT-SCC-9 (UT-SCC-9P) as well as its cetuximab (CTX)-resistant sub-clone (UT-SCC-9R) were forced to grow in an anchorage-independent manner by coating culture dishes with the anti-adhesive polymer poly-2-hydroxyethylmethacrylate (poly-HEMA). The extent of apoptosis, clonogenic survival and EGFR signalling under such culture conditions was evaluated. The potential of spheroid formation in suspension culture was found to be positively correlated with the proliferation rate of HNSCC cell lines as well as their basal EGFR expression levels. CTX and gefitinib blocked, whereas the addition of EGFR ligands promoted anchorage-independent cell survival and spheroid formation. Increased spheroid formation and growth were associated with persistent activation of EGFR and its downstream signalling component (MAPK/ERK). Importantly, HNSCC cells derived from spheroid cultures retained their clonogenic potential in the absence of cell-matrix contact. Addition of CTX under these conditions strongly inhibited colony formation in CTX-sensitive cell lines but not their resistant subclones. Altogether, EGFR activation was identified as crucial factor for anchorage-independent survival of HNSCC cells. Targeting EGFR in CTC cluster formation might represent an attractive anti-metastatic treatment approach in HNSCC.
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DOI：
10.1186/bcr2349
发表时间：
2009
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Fehm T;Hoffmann O;Aktas B;Becker S;Solomayer EF;Wallwiener D;Kimmig R;Kasimir-Bauer S
通讯作者：
Kasimir-Bauer S
影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
影响因子：
11.5
作者：
Hayes, Daniel F.;Cristofanilli, Massimo;Terstappen, Leon W. W. M.
通讯作者：
Terstappen, Leon W. W. M.
影响因子：
28.2
作者：
Bardelli A;Corso S;Bertotti A;Hobor S;Valtorta E;Siravegna G;Sartore-Bianchi A;Scala E;Cassingena A;Zecchin D;Apicella M;Migliardi G;Galimi F;Lauricella C;Zanon C;Perera T;Veronese S;Corti G;Amatu A;Gambacorta M;Diaz LA Jr;Sausen M;Velculescu VE;Comoglio P;Trusolino L;Di Nicolantonio F;Giordano S;Siena S
通讯作者：
Siena S
影响因子：
3.8
作者：
Gorges TM;Tinhofer I;Drosch M;Röse L;Zollner TM;Krahn T;von Ahsen O
通讯作者：
von Ahsen O