Simulating tumor microenvironment: changes in protein expression in an in vitro co-culture system

Cancer Cell International

Mirella Falconi¹  Maria Carla Mazzotti¹  Sandra Durante¹  Stefano Focaroli¹  Silvia Bolzani¹  Gabriella Teti¹  Viviana Salvatore¹ 

[1] Department for Biomedical and Neuromotor Sciences (DIBINEM), University of Bologna, via Irnerio 48, Bologna 40126, Italy

关键词: Human fibroblast cells;    Osteosarcoma cell line;    MMP1;    VEGF;    YKL-40;    Cell-cell contacts;    Co-cultures;    Tumor microenvironment;   

Others  :  792020
DOI  :  10.1186/1475-2867-14-40

 received in 2013-11-21, accepted in 2014-05-01,  发布年份 2014

【 摘 要 】

Background

The role of the microenvironment during the initiation and progression of carcinogenesis is thought to be of critical importance, both for the enhanced understanding of fundamental cancer biology as well as for improving molecular diagnostics and therapeutics. The aim of this study was to establish an in vitro model based on a co-culture of healthy human fibroblasts (HFs) and human osteosarcoma cells (MG-63s) to simulate the microenvironment including tumor and healthy cells.

Methods

The HFs and MG-63s were in vitro co-cultured for a period of time ranging from 24 h to 7 days. Cell morphology and organization were studied using phase contrast microscopy while the expression of Human Cartilage Glycoprotein 39 (YKL-40), Vascular Endothelial Growth Factor (VEGF) and Matrix Metalloprotease 1 (MMP1) was investigated by Real Time PCR and Western Blotting.

Results

The results showed a characteristic disposition of tumor and healthy co-cultured cells in columns which are not visible in tumor and healthy cells grown singularly. The expression of YKL-40, VEGF and MMP1 significantly changed in co-cultured cells compared to HFs and MG-63s separately cultured.

Conclusions

We concluded that the tumor microenvironment has an influence on the protein expression of the healthy surrounding tissues and the process of tumorigenicity.

【 授权许可】

   
2014 Salvatore et al.; licensee BioMed Central Ltd.

【 图 表 】

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