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The primary growth of laryngeal squamous cell carcinoma cells in vitro is effectively supported by paired cancer-associated fibroblasts alone
- Source :
- Tumor Biology, Vol 39 (2017)
- Publication Year :
- 2017
- Publisher :
- IOS Press, 2017.
-
Abstract
- Most primarily cultured laryngeal squamous cell carcinoma cells are difficult to propagate in vitro and have a low survival rate. However, in our previous work to establish a laryngeal squamous cell carcinoma cell line, we found that laryngeal cancer-associated fibroblasts appeared to strongly inhibit the apoptosis of primarily cultured laryngeal squamous cell carcinoma cells in vitro. In this study, we investigated whether paired laryngeal cancer-associated fibroblasts alone can effectively support the growth of primarily cultured laryngeal squamous cell carcinoma cells in vitro. In all, 29 laryngeal squamous cell carcinoma specimens were collected and primarily cultured. The laryngeal squamous cell carcinoma cells were separated from cancer-associated fibroblasts by differential trypsinization and continuously subcultured. Morphological changes of the cultured laryngeal squamous cell carcinoma cells were observed. Immunocytofluorescence was used to authenticate the identity of the cancer-associated fibroblasts and laryngeal squamous cell carcinoma cells. Flow cytometry was used to quantify the proportion of apoptotic cells. Western blot was used to detect the protein levels of caspase-3. Enzyme-linked immunosorbent assay was used to detect the levels of chemokine (C-X-C motif) ligand 12, chemokine (C-X-C motif) ligand 7, hepatocyte growth factor, and fibroblast growth factor 1 in the supernatants of the laryngeal squamous cell carcinoma and control cells. AMD3100 (a chemokine (C-X-C motif) receptor 4 antagonist) and an anti–chemokine (C-X-C motif) ligand 7 antibody were used to block the tumor-supporting capacity of cancer-associated fibroblasts. Significant apoptotic changes were detected in the morphology of laryngeal squamous cell carcinoma cells detached from cancer-associated fibroblasts. The percentage of apoptotic laryngeal squamous cell carcinoma cells and the protein levels of caspase-3 increased gradually in subsequent subcultures. In contrast, no significant differences in the proliferation capacity of laryngeal squamous cell carcinoma cells cocultured with cancer-associated fibroblasts were detected during subculturing. High level of chemokine (C-X-C motif) ligand 12 was detected in the culture supernatant of cancer-associated fibroblasts. The tumor-supporting effect of cancer-associated fibroblasts was significantly inhibited by AMD3100. Our findings demonstrate that the paired laryngeal cancer-associated fibroblasts alone are sufficient to support the primary growth of laryngeal squamous cell carcinoma cells in vitro and that the chemokine (C-X-C motif) ligand 12/chemokine (C-X-C motif) receptor 4 axis is one of the major contributors.
- Subjects :
- Male
0301 basic medicine
Benzylamines
Receptors, CXCR4
Chemokine
Pathology
medicine.medical_specialty
Apoptosis
Biology
Cyclams
03 medical and health sciences
0302 clinical medicine
Cancer-Associated Fibroblasts
Heterocyclic Compounds
medicine
Humans
Stromal cell-derived factor 1
Chemokine CCL7
Laryngeal Neoplasms
RC254-282
Cell Proliferation
Neoplasm Staging
Hepatocyte Growth Factor
Cell growth
Neoplasms. Tumors. Oncology. Including cancer and carcinogens
General Medicine
Laryngeal Neoplasm
Flow Cytometry
Chemokine CXCL12
Trypsinization
Gene Expression Regulation, Neoplastic
030104 developmental biology
Cell culture
030220 oncology & carcinogenesis
Carcinoma, Squamous Cell
Cancer research
biology.protein
Fibroblast Growth Factor 1
Hepatocyte growth factor
medicine.drug
Subjects
Details
- ISSN :
- 14230380 and 10104283
- Volume :
- 39
- Database :
- OpenAIRE
- Journal :
- Tumor Biology
- Accession number :
- edsair.doi.dedup.....47660a4345698f9388e568d4d2edfbf9