HDGF enhances VEGFdependent angiogenesis and FGF2 is a VEGFindependent angiogenic factor in nonsmall cell lung cancer.
Oncol Rep
; 44(1): 14-28, 2020 07.
Article
in En
| MEDLINE
| ID: mdl-32319650
ABSTRACT
Nonsmall cell lung cancer (NSCLC) accounts for over 80% of all diagnosed lung cancer cases. Lung cancer is the leading cause of cancerrelated deaths worldwide. Most NSCLC cells overexpress vascular endothelial growth factorA (VEGFA) which plays a pivotal role in tumour angiogenesis. Antiangiogenic therapies including VEGFA neutralisation have significantly improved the response rates, progressionfree survival and overall survival of patients with NSCLC. However, the median survival of these patients is shorter than 18 months, suggesting that NSCLC cells secrete VEGFindependent angiogenic factors, which remain unknown. We aimed to explore these factors in human NSCLC cell lines, A549, Lu99 and EBC1 using serumfree culture, to which only EBC1 cells could adapt. By mass spectrometry, we identified 1,007 proteins in the culture supernatant derived from EBC1 cells. Among the identified proteins, interleukin8 (IL8), macrophage migration inhibitory factor (MIF), galectin1, midkine (MK), IL18, galectin3, VEGFA, hepatomaderived growth factor (HDGF), osteopontin (OPN), connective tissue growth factor (CTGF) and granulin (GRN) are known to be involved in angiogenesis. Tube formation, neutralisation and RNA interference assays revealed that VEGFA and HDGF function as angiogenic factors in EBC1 cells. To confirm whether VEGFA and HDGF also regulate angiogenesis in the other NSCLC cell lines, we established a novel culture method. NSCLC cells were embedded in collagen gel and cultured threedimensionally. Tube formation, neutralisation and RNA interference assays using the threedimensional (3D) culture supernatant showed that VEGFA and HDGF were not angiogenic factors in Lu99 cells. By gene microarray in EBC1 and Lu99 cells, we identified 61 mRNAs expressed only in Lu99 cells. Among these mRNAs, brainderived neurotrophic factor (BDNF), fibroblast growth factor2 (FGF2) and FGF5 are known to be involved in angiogenesis. Tube formation and neutralisation assays clarified that FGF2 functions as an angiogenic factor in Lu99 cells. These results indicate that HDGF enhances VEGFdependent angiogenesis and that FGF2 is a VEGFindependent angiogenic factor in human NSCLC cells.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Fibroblast Growth Factor 2
/
Culture Media, Serum-Free
/
Carcinoma, Non-Small-Cell Lung
/
Intercellular Signaling Peptides and Proteins
/
Vascular Endothelial Growth Factor A
/
Lung Neoplasms
Type of study:
Prognostic_studies
Limits:
Humans
Language:
En
Journal:
Oncol Rep
Journal subject:
NEOPLASIAS
Year:
2020
Type:
Article
Affiliation country:
Japan