Potential role for Ext1-dependent heparan sulfate in regulating P311 gene expression in A549 carcinoma cells
| Autor: | Lawrence Fred Sembajwe, Kirankumar Katta, Marion Kusche-Gullberg |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Small interfering RNA Stromal cell medicine.medical_treatment Biophysics Nerve Tissue Proteins N-Acetylglucosaminyltransferases Biochemistry Transforming Growth Factor beta1 03 medical and health sciences chemistry.chemical_compound Mice Gene expression medicine Animals Humans Fibroblast Molecular Biology Cell Proliferation A549 cell Chemistry Growth factor Gene Expression Profiling Heparan sulfate Fibroblasts Embryo Mammalian Coculture Techniques Cell biology Gene expression profiling 030104 developmental biology medicine.anatomical_structure Gene Expression Regulation A549 Cells Heparitin Sulfate |
| Zdroj: | Biochimica et biophysica acta. General subjects. 1862(6) |
| ISSN: | 0304-4165 |
| Popis: | Background Exostosin-1 (EXT1), a member of the EXT protein family, is indispensable for synthesis of heparan sulfate (HS) chains that bind to and modulate the signaling efficiency of numerous growth factor activities. We have previously shown that Ext1 mutated mouse embryonic fibroblasts produce short sulfated HS chains which dramatically influence tumor cell behavior in a 3-dimensional (3D) heterospheroid system composed of tumor cells and fibroblasts. Methods In this study, we have used both 2D co-culture and 3D heterospheroid models, consisting of human A549 carcinoma cells co-cultured with wild-type or Ext1-mutated mouse embryonic fibroblasts. Results and conclusions Gene expression profiling of differentially expressed genes in fibroblast/A549 heterospheroids identified P311 as a gene substantially down-regulated in A549 cells co-cultured with Ext1-mutated fibroblasts. In addition, we observed that the Ext1 mutants displayed reduced Tgf-β1 mRNA levels and lower levels of secreted active TGF-β protein. Re-introduction of Ext1 in the Ext1 mutant fibroblasts rescued the levels of Tgf-β1 mRNA, increased the amounts of secreted active TGF-β in these cells, as well as P311 mRNA levels in adjacent A549 cells. Accordingly, small interfering RNAs (siRNAs) against fibroblast Tgf-β1 reduced P311 expression in neighboring A549 tumor cells. Our data raises the possibility that fibroblast Ext1 levels play a role in P311 expression in A549/fibroblast co-culture through TGF-β1. General significance This study considers a possible novel mechanism of Ext1-regulated heparan sulfate structure in modifying tumor-stroma interactions through altering stromal tgf-s1 expression. |
| Databáze: | OpenAIRE |
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