ETHIANUM - Clinic for Plastic, Aesthetic and Reconstructive Surgery, Spine, Orthopedic and Hand Surgery, Preventive Medicine, Voßstraße 6, 69115, Heidelberg, Germany.
Department of Medical Oncology, National Center for Tumor Diseases (NCT) Heidelberg, Heidelberg University Hospital, Im Neuenheimer Feld 460, 60120, Heidelberg, Germany.
Stem Cell Rev Rep. 2018 Feb;14(1):125-140. doi: 10.1007/s12015-017-9772-y.
This study focuses on the interactions of human adipose tissue-derived stem cells (ADSCs) and malignant melanoma cells (MMCs) with regard to future cell-based skin therapies. The aim was to identify potential oncological risks as ADSCs could unintentionally be sited within the proximity of the tumor microenvironment of MMCs. An indirect co-culture model was used to analyze interactions between ADSCs and four different established melanoma cell lines (G-361, SK-Mel-5, MeWo and A2058) as well as two low-passage primary melanoma cell cultures (M1 and M2). Doubling time, migration and invasion, angiogenesis, quantitative real-time PCR of 229 tumor-associated genes and multiplex protein assays of 20 chemokines and growth factors and eight matrix metalloproteinases (MMPs) were evaluated. Co-culture with ADSCs significantly increased migration capacity of G-361, SK-Mel-5, A2058, MeWo and M1 and invasion capacity of G-361, SK-Mel-5 and A2058 melanoma cells. Furthermore, conditioned media from all ADSC-MMC-co-cultures induced tube formation in an angiogenesis assay in vitro. Gene expression analysis of ADSCs and MMCs, especially of low-passage melanoma cell cultures, revealed an increased expression of various genes with tumor-promoting activities, such as CXCL12, PTGS2, IL-6, and HGF upon ADSC-MMC-co-culture. In this context, a significant increase (up to 5,145-fold) in the expression of numerous tumor-associated proteins could be observed, e.g. several pro-angiogenic factors, such as VEGF, IL-8, and CCL2, as well as different matrix metalloproteinases, especially MMP-2. In conclusion, the current report clearly demonstrates that a bi-directional crosstalk between ADSCs and melanoma cells can enhance different malignant properties of melanoma cells in vitro.
本研究关注人脂肪组织源性干细胞（ADSCs）与恶性黑色素瘤细胞（MMCs）之间的相互作用，以期发现未来基于细胞的皮肤治疗的潜在肿瘤风险。由于 ADSC 可能会意外位于 MMC 肿瘤微环境附近，因此我们的目的是识别这些风险。我们使用间接共培养模型分析了 ADSC 与四种不同的已建立的黑色素瘤细胞系（G-361、SK-Mel-5、MeWo 和 A2058）以及两种低传代原发性黑色素瘤细胞培养物（M1 和 M2）之间的相互作用。我们评估了倍增时间、迁移和侵袭、血管生成、229 个肿瘤相关基因的定量实时 PCR 和 20 种趋化因子和生长因子以及 8 种基质金属蛋白酶（MMPs）的多重蛋白分析。与 ADSC 的共培养显著增加了 G-361、SK-Mel-5、A2058、MeWo 和 M1 的迁移能力，以及 G-361、SK-Mel-5 和 A2058 黑色素瘤细胞的侵袭能力。此外，来自所有 ADSC-MMC 共培养物的条件培养基在体外血管生成测定中诱导了管形成。ADSC 和 MMC 的基因表达分析，特别是低传代黑色素瘤细胞培养物的分析，揭示了在 ADSC-MMC 共培养后，各种具有促肿瘤活性的基因的表达增加，例如 CXCL12、PTGS2、IL-6 和 HGF。在这方面，观察到许多肿瘤相关蛋白的表达显著增加（高达 5145 倍），例如多种促血管生成因子，如 VEGF、IL-8 和 CCL2，以及不同的基质金属蛋白酶，特别是 MMP-2。总之，本报告清楚地表明，ADSC 与黑色素瘤细胞之间的双向串扰可以增强黑色素瘤细胞在体外的不同恶性特性。
