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Circulating tumor cells from 4D model have less integrin beta 4 expression
Dhruva K Mishra1, Kenneth L Scott2, Joanna M Wardwell-Ozgo2
1Department of Surgery, Houston Methodist Research Institute, Houston, Texas.
Background:
Currently, there is no in vitro or ex vivo model that can isolate circulating tumor cells (CTCs). Recently, we developed a four-dimensional (4D) lung cancer model that allows for the isolation of CTCs. We postulated that these cells have different properties than parental (2D) cells.
Materials And Methods:
We obtained CTCs by growing A549, H1299, 393P, and 344SQ cell lines on the 4D lung model. The CTCs were functionally characterized in vitro and gene expression of the cell adhesion molecules was compared with respective 2D cells. Integrin beta 4 (ITGB4) was further investigated by stably transfecting the A549 and H1299 cells.
Results:
We found that all cell lines produced CTCs, and that CTCs from the 4D model were less adherent to the plastic and have a slower growth rate than respective 2D cells (P < 0.01). Most of the cell adhesion molecules were downregulated (P < 0.05) in CTCs, and ITGB4 was the common molecule, significantly more underexpressed in CTCs from all cell lines than their respective 2D cells. The modulation of ITGB4 led to a differential function of 2D cells.
Conclusions:
CTCs from the 4D model have different transcriptional, translational, and in vitro characteristics than the same cells grown on a petri dish, and these CTCs from the 4D model have the properties of CTCs that are responsible for metastasis.
Insights
A novel four-dimensional (4D) lung cancer model successfully isolates circulating tumor cells (CTCs). These 4D-derived CTCs exhibit distinct properties, including reduced adhesion and slower growth, crucial for metastasis.
Area of Science:
- Oncology
- Cell Biology
- Biotechnology
Background:
- Limited in vitro/ex vivo models for isolating circulating tumor cells (CTCs).
- Development of a novel four-dimensional (4D) lung cancer model for CTC isolation.
- Hypothesis that 4D-derived CTCs possess different properties than traditional 2D cultured cells.
Purpose of the Study:
- To characterize circulating tumor cells (CTCs) isolated using a 4D lung cancer model.
- To compare the properties of CTCs derived from the 4D model with parental 2D cells.
- To investigate the role of cell adhesion molecules, specifically Integrin beta 4 (ITGB4), in CTC behavior.
Main Methods:
- Generated CTCs from A549, H1299, 393P, and 344SQ cell lines using the 4D lung model.
- Performed in vitro functional characterization of CTCs.
- Compared gene expression of cell adhesion molecules between 4D-derived CTCs and 2D cells.
- Investigated ITGB4 function through stable transfection in A549 and H1299 cells.
Main Results:
- All tested cell lines produced CTCs using the 4D model.
- CTCs from the 4D model showed significantly reduced adherence and slower growth rates compared to 2D cells (P < 0.01).
- Most cell adhesion molecules were downregulated in CTCs (P < 0.05), with ITGB4 being significantly underexpressed across all cell lines.
- Modulation of ITGB4 resulted in differential functional changes in 2D cells.
Conclusions:
- CTCs derived from the 4D lung model exhibit distinct transcriptional, translational, and in vitro characteristics compared to 2D cultured cells.
- The 4D-derived CTCs possess properties associated with metastatic potential.
- The 4D model provides a valuable tool for studying CTCs and their role in cancer metastasis.
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