TY - JOUR
T1 - The Formation of Stable Lung Tumor Spheroids during Random Positioning Involves Increased Estrogen Sensitivity
AU - Barkia, Balkis
AU - Sandt, Viviann
AU - Melnik, Daniela
AU - Cortés-Sánchez, José Luis
AU - Marchal, Shannon
AU - Baselet, Bjorn
AU - Baatout, Sarah
AU - Sahana, Jayashree
AU - Grimm, Daniela
AU - Wehland, Markus
AU - Schulz, Herbert
AU - Infanger, Manfred
AU - Kraus, Armin
AU - Krüger, Marcus
PY - 2024/10/12
Y1 - 2024/10/12
N2 - The formation of tumor spheroids on the random positioning machine (RPM) is a complex and important process, as it enables the study of metastasis ex vivo. However, this process is not yet understood in detail. In this study, we compared the RPM-induced spheroid formation of two cell types of lung carcinoma (NCI-H1703 squamous cell carcinoma cells and Calu-3 adenocarcinoma cells). While NCI-H1703 cells were mainly present as spheroids after 3 days of random positioning, Calu-3 cells remained predominantly as a cell layer. We found that two-dimensional-growing Calu-3 cells have less mucin-1, further downregulate their expression on the RPM and therefore exhibit a higher adhesiveness. In addition, we observed that Calu-3 cells can form spheroids, but they are unstable due to an imbalanced ratio of adhesion proteins (β1-integrin, E-cadherin) and anti-adhesion proteins (mucin-1) and are likely to disintegrate in the shear environment of the RPM. RPM-exposed Calu-3 cells showed a strongly upregulated expression of the estrogen receptor alpha gene ESR1. In the presence of 17β-estradiol or phenol red, more stable Calu-3 spheroids were formed, which was presumably related to an increased amount of E-cadherin in the cell aggregates. Thus, RPM-induced tumor spheroid formation depends not solely on cell-type-specific properties but also on the complex interplay between the mechanical influences of the RPM and, to some extent, the chemical composition of the medium used during the experiments.
AB - The formation of tumor spheroids on the random positioning machine (RPM) is a complex and important process, as it enables the study of metastasis ex vivo. However, this process is not yet understood in detail. In this study, we compared the RPM-induced spheroid formation of two cell types of lung carcinoma (NCI-H1703 squamous cell carcinoma cells and Calu-3 adenocarcinoma cells). While NCI-H1703 cells were mainly present as spheroids after 3 days of random positioning, Calu-3 cells remained predominantly as a cell layer. We found that two-dimensional-growing Calu-3 cells have less mucin-1, further downregulate their expression on the RPM and therefore exhibit a higher adhesiveness. In addition, we observed that Calu-3 cells can form spheroids, but they are unstable due to an imbalanced ratio of adhesion proteins (β1-integrin, E-cadherin) and anti-adhesion proteins (mucin-1) and are likely to disintegrate in the shear environment of the RPM. RPM-exposed Calu-3 cells showed a strongly upregulated expression of the estrogen receptor alpha gene ESR1. In the presence of 17β-estradiol or phenol red, more stable Calu-3 spheroids were formed, which was presumably related to an increased amount of E-cadherin in the cell aggregates. Thus, RPM-induced tumor spheroid formation depends not solely on cell-type-specific properties but also on the complex interplay between the mechanical influences of the RPM and, to some extent, the chemical composition of the medium used during the experiments.
KW - Cadherins/metabolism
KW - Cell Adhesion/drug effects
KW - Cell Line, Tumor
KW - Estradiol/pharmacology
KW - Estrogen Receptor alpha/metabolism
KW - Estrogens/pharmacology
KW - Gene Expression Regulation, Neoplastic/drug effects
KW - Humans
KW - Integrin beta1/metabolism
KW - Lung Neoplasms/metabolism
KW - Mucin-1/metabolism
KW - Spheroids, Cellular/metabolism
KW - tumor spheroids
KW - lung cancer
KW - phenol red
KW - cell signaling
KW - simulated microgravity
UR - http://www.scopus.com/inward/record.url?scp=85207681117&partnerID=8YFLogxK
U2 - 10.3390/biom14101292
DO - 10.3390/biom14101292
M3 - Journal article
C2 - 39456226
SN - 2218-273X
VL - 14
JO - Biomolecules
JF - Biomolecules
IS - 10
M1 - 1292
ER -