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Updated: Mar 30, 2026
A Cancer Cell Spheroid Assay to Assess Invasion in a 3D Setting
Published on: November 20, 2015
JAK/STAT pathway interacts with intercellular cell adhesion molecule (ICAM) and vascular cell adhesion molecule
Fahriye Duzagac1, Sevinc Inan, Fatma Ela Simsek
1Department of Stem Cell, Institute of Health Sciences, Ege University, Izmir, Turkey.
Purpose:
JAK/STAT is an evolutionarily conserved pathway and very important for second messenger system. This pathway is important in malignant transformation and accumulated evidence indicates that this pathway is involved in tumorigenesis and progression of several cancers. It was possible to assume that activation of JAK/STAT pathway is associated with increase in the expressions of ICAM/1 and VCAM-1. In this study we hypothesized that when cells were maintained as spheroids or monolayers, the structure of cancer stem cells (CSCs) could show differentiation when compared with non-CSCs.
Methods:
DU-145 human prostate cancer cells were cultured using the Ege University molecular embryology laboratory medium supplemented wıth 10% fetal bovine serum. Clusters of differentiation 133 (CD133)(+high)/CD44(+high) prostate CSCs were isolated from the DU145 cell line by using BD FACSAria. CD133//CD44+ CSCs were cultured until confluent with 3% noble agar. The expression of these proteins in CSCs and non-CSCs was analyzed by immunohistochemistry.
Results:
Different expression profiles were observed in the conventional two-dimensional (2D) and three-dimensional (3D) experimental model system when CSCs and non-CSCs were compared. Human prostate CSCs exhibited intense ICAM-1 and VCAM-1 immunoreaction when compared with non-CSCs. These findings were supported by the fact that VCAM-1 on the surface of cancer cells binds to its counterreceptor, the α4β1 integrin (also known as very-late antigen, VLA-4), on metastasis-associated macrophages, triggering VCAM-1-mediated activation of the phosphoinositide 3-kinase growth and survival pathway in cancer cells.
Conclusions:
The results of this study showed that changes in JAK/STAT pathway are related with adhesion molecules and could affect cancer progression.
Insights
Cancer stem cells (CSCs) show increased expression of adhesion molecules ICAM-1 and VCAM-1, linked to the JAK/STAT pathway. This suggests a connection between JAK/STAT signaling, adhesion molecules, and cancer progression.
Area of Science:
- Cell Biology
- Cancer Research
- Molecular Biology
Background:
- The Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway is crucial for cellular communication and implicated in cancer development.
- Understanding the role of cancer stem cells (CSCs) in tumorigenesis is vital for developing effective cancer therapies.
Purpose of the Study:
- To investigate the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in prostate cancer stem cells (CSCs) compared to non-CSCs.
- To explore the potential association between the JAK/STAT pathway, adhesion molecules, and cancer stem cell differentiation.
Main Methods:
- Prostate CSCs (CD133+/CD44+) were isolated from DU-145 cells using flow cytometry.
- CSCs and non-CSCs were cultured in 2D and 3D models and analyzed for protein expression via immunohistochemistry.
Main Results:
- Prostate CSCs displayed significantly higher ICAM-1 and VCAM-1 immunoreactivity than non-CSCs.
- VCAM-1 expression on cancer cells correlated with α4β1 integrin on macrophages, activating pro-survival pathways.
Conclusions:
- Alterations in the JAK/STAT pathway are associated with changes in adhesion molecules.
- These molecular changes may influence cancer progression and highlight potential therapeutic targets.
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