Department of Pathology, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands.
Br J Cancer. 2013 Sep 3;109(5):1214-22. doi: 10.1038/bjc.2013.451. Epub 2013 Aug 6.
Chondrosarcomas are malignant cartilage-forming tumours of bone. Because of their resistance to conventional chemotherapy and radiotherapy, currently no treatment strategies exist for unresectable and metastatic chondrosarcoma. Previously, PI3K/AKT/GSK3β and Src kinase pathways were shown to be activated in chondrosarcoma cell lines. Our aim was to investigate the role of these kinases in chemoresistance and migration in chondrosarcoma in relation to TP53 mutation status.
We used five conventional and three dedifferentiated chondrosarcoma cell lines and investigated the effect of PI3K/AKT/GSK3β pathway inhibition (enzastaurin) and Src pathway inhibition (dasatinib) in chemoresistance using WST assay and live cell imaging with AnnexinV staining. Immunohistochemistry on tissue microarrays (TMAs) containing 157 cartilaginous tumours was performed for Src family members. Migration assays were performed with the RTCA xCelligence System.
Src inhibition was found to overcome chemoresistance, to induce apoptosis and to inhibit migration. Cell lines with TP53 mutations responded better to combination therapy than wild-type cell lines (P=0.002). Tissue microarray immunohistochemistry confirmed active Src (pSrc) signalling, with Fyn being most abundantly expressed (76.1%).
These results strongly indicate Src family kinases, in particular Fyn, as a potential target for the treatment of inoperable and metastatic chondrosarcomas, and to sensitise for doxorubicin especially in the presence of TP53 mutations.
软骨肉瘤是一种恶性的软骨形成性骨肿瘤。由于其对常规化疗和放疗的抵抗力，目前对于无法切除和转移性软骨肉瘤尚无治疗策略。先前的研究表明，PI3K/AKT/GSK3β 和Src 激酶途径在软骨肉瘤细胞系中被激活。我们的目的是研究这些激酶在与 TP53 突变状态相关的软骨肉瘤中的化学抗性和迁移中的作用。
我们使用了五株常规和三株去分化软骨肉瘤细胞系，并使用 WST 测定法和 AnnexinV 染色的活细胞成像来研究 PI3K/AKT/GSK3β 途径抑制（恩杂鲁胺）和 Src 途径抑制（达沙替尼）对化学抗性的影响。对包含 157 个软骨肿瘤的组织微阵列（TMA）进行了Src 家族成员的免疫组织化学染色。使用 RTCA xCelligence 系统进行迁移测定。
发现 Src 抑制可克服化学抗性，诱导细胞凋亡并抑制迁移。具有 TP53 突变的细胞系对联合治疗的反应优于野生型细胞系（P=0.002）。组织微阵列免疫组织化学证实了活跃的Src（pSrc）信号传导，其中 Fyn 的表达最为丰富（76.1%）。
这些结果强烈表明 Src 家族激酶，特别是 Fyn，是治疗不可切除和转移性软骨肉瘤的潜在靶标，并在存在 TP53 突变的情况下增强对多柔比星的敏感性。
