Long-Term Simulation of Microgravity Induces Changes in Gene Expression in Breast Cancer Cells
Microgravity changes the gene expression pattern in various cell types. This study focuses on the breast cancer cell lines MCF-7 (less invasive) and MDA-MB-231 (triple-negative, highly invasive). The cells were cultured for 14 days under simulated microgravity (s-µ<i>g</i>) conditions us...
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| Format: | Article |
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MDPI AG
2023-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/2/1181 |
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| author | Jayashree Sahana José Luis Cortés-Sánchez Viviann Sandt Daniela Melnik Thomas J. Corydon Herbert Schulz Zexi Cai Katja Evert Daniela Grimm Markus Wehland |
| author_facet | Jayashree Sahana José Luis Cortés-Sánchez Viviann Sandt Daniela Melnik Thomas J. Corydon Herbert Schulz Zexi Cai Katja Evert Daniela Grimm Markus Wehland |
| author_sort | Jayashree Sahana |
| collection | DOAJ |
| description | Microgravity changes the gene expression pattern in various cell types. This study focuses on the breast cancer cell lines MCF-7 (less invasive) and MDA-MB-231 (triple-negative, highly invasive). The cells were cultured for 14 days under simulated microgravity (s-µ<i>g</i>) conditions using a random positioning machine (RPM). We investigated cytoskeletal and extracellular matrix (ECM) factors as well as focal adhesion (FA) and the transmembrane proteins involved in different cellular signaling pathways (MAPK, PAM and VEGF). The mRNA expressions of 24 genes of interest (<i>TUBB</i>, <i>ACTB</i>, <i>COL1A1</i>, <i>COL4A5</i>, <i>LAMA3</i>, <i>ITGB1</i>, <i>CD44</i>, <i>VEGF</i>, <i>FLK1</i>, <i>EGFR</i>, <i>SRC</i>, <i>FAK1</i>, <i>RAF1</i>, <i>AKT1</i>, <i>ERK1</i>, <i>MAPK14</i>, <i>MAP2K1</i>, <i>MTOR</i>, <i>RICTOR</i>, <i>VCL</i>, <i>PXN</i>, <i>CDKN1</i>, <i>CTNNA1</i> and <i>CTNNB1</i>) were determined by quantitative real-time PCR (qPCR) and studied using STRING interaction analysis. Histochemical staining was carried out to investigate the morphology of the adherent cells (ADs) and the multicellular spheroids (MCSs) after RPM exposure. To better understand this experimental model in the context of breast cancer patients, a weighted gene co-expression network analysis (WGCNA) was conducted to obtain the expression profiles of 35 breast cell lines from the HMS LINCS Database. The qPCR-verified genes were searched in the mammalian phenotype database and the human genome-wide association studies (GWAS) Catalog. The results demonstrated the positive association between the real metastatic microtumor environment and MCSs with respect to the extracellular matrix, cytoskeleton, morphology, different cellular signaling pathway key proteins and several other components. In summary, the microgravity-engineered three-dimensional MCS model can be utilized to study breast cancer cell behavior and to assess the therapeutic efficacies of drugs against breast cancer in the future. |
| first_indexed | 2024-03-09T12:25:14Z |
| format | Article |
| id | doaj.art-d72e030a85844752a30363a0b37e6e3f |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T12:25:14Z |
| publishDate | 2023-01-01 |
| publisher | MDPI AG |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d72e030a85844752a30363a0b37e6e3f2023-11-30T22:36:35ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-01-01242118110.3390/ijms24021181Long-Term Simulation of Microgravity Induces Changes in Gene Expression in Breast Cancer CellsJayashree Sahana0José Luis Cortés-Sánchez1Viviann Sandt2Daniela Melnik3Thomas J. Corydon4Herbert Schulz5Zexi Cai6Katja Evert7Daniela Grimm8Markus Wehland9Department of Biomedicine, Aarhus University, 8000 Aarhus, DenmarkDepartment of Microgravity and Translational Regenerative Medicine, Otto von Guericke University, 39106 Magdeburg, GermanyDepartment of Microgravity and Translational Regenerative Medicine, Otto von Guericke University, 39106 Magdeburg, GermanyDepartment of Microgravity and Translational Regenerative Medicine, Otto von Guericke University, 39106 Magdeburg, GermanyDepartment of Biomedicine, Aarhus University, 8000 Aarhus, DenmarkDepartment of Microgravity and Translational Regenerative Medicine, Otto von Guericke University, 39106 Magdeburg, GermanyCenter for Quantitative Genetics and Genomics, Aarhus University, 8000 Aarhus, DenmarkInstitute of Pathology, University of Regensburg, 93953 Regensburg, GermanyDepartment of Biomedicine, Aarhus University, 8000 Aarhus, DenmarkDepartment of Microgravity and Translational Regenerative Medicine, Otto von Guericke University, 39106 Magdeburg, GermanyMicrogravity changes the gene expression pattern in various cell types. This study focuses on the breast cancer cell lines MCF-7 (less invasive) and MDA-MB-231 (triple-negative, highly invasive). The cells were cultured for 14 days under simulated microgravity (s-µ<i>g</i>) conditions using a random positioning machine (RPM). We investigated cytoskeletal and extracellular matrix (ECM) factors as well as focal adhesion (FA) and the transmembrane proteins involved in different cellular signaling pathways (MAPK, PAM and VEGF). The mRNA expressions of 24 genes of interest (<i>TUBB</i>, <i>ACTB</i>, <i>COL1A1</i>, <i>COL4A5</i>, <i>LAMA3</i>, <i>ITGB1</i>, <i>CD44</i>, <i>VEGF</i>, <i>FLK1</i>, <i>EGFR</i>, <i>SRC</i>, <i>FAK1</i>, <i>RAF1</i>, <i>AKT1</i>, <i>ERK1</i>, <i>MAPK14</i>, <i>MAP2K1</i>, <i>MTOR</i>, <i>RICTOR</i>, <i>VCL</i>, <i>PXN</i>, <i>CDKN1</i>, <i>CTNNA1</i> and <i>CTNNB1</i>) were determined by quantitative real-time PCR (qPCR) and studied using STRING interaction analysis. Histochemical staining was carried out to investigate the morphology of the adherent cells (ADs) and the multicellular spheroids (MCSs) after RPM exposure. To better understand this experimental model in the context of breast cancer patients, a weighted gene co-expression network analysis (WGCNA) was conducted to obtain the expression profiles of 35 breast cell lines from the HMS LINCS Database. The qPCR-verified genes were searched in the mammalian phenotype database and the human genome-wide association studies (GWAS) Catalog. The results demonstrated the positive association between the real metastatic microtumor environment and MCSs with respect to the extracellular matrix, cytoskeleton, morphology, different cellular signaling pathway key proteins and several other components. In summary, the microgravity-engineered three-dimensional MCS model can be utilized to study breast cancer cell behavior and to assess the therapeutic efficacies of drugs against breast cancer in the future.https://www.mdpi.com/1422-0067/24/2/1181breast cancerspheroidscytoskeletonextracellular matrixfocal adhesionmicrogravity |
| spellingShingle | Jayashree Sahana José Luis Cortés-Sánchez Viviann Sandt Daniela Melnik Thomas J. Corydon Herbert Schulz Zexi Cai Katja Evert Daniela Grimm Markus Wehland Long-Term Simulation of Microgravity Induces Changes in Gene Expression in Breast Cancer Cells International Journal of Molecular Sciences breast cancer spheroids cytoskeleton extracellular matrix focal adhesion microgravity |
| title | Long-Term Simulation of Microgravity Induces Changes in Gene Expression in Breast Cancer Cells |
| title_full | Long-Term Simulation of Microgravity Induces Changes in Gene Expression in Breast Cancer Cells |
| title_fullStr | Long-Term Simulation of Microgravity Induces Changes in Gene Expression in Breast Cancer Cells |
| title_full_unstemmed | Long-Term Simulation of Microgravity Induces Changes in Gene Expression in Breast Cancer Cells |
| title_short | Long-Term Simulation of Microgravity Induces Changes in Gene Expression in Breast Cancer Cells |
| title_sort | long term simulation of microgravity induces changes in gene expression in breast cancer cells |
| topic | breast cancer spheroids cytoskeleton extracellular matrix focal adhesion microgravity |
| url | https://www.mdpi.com/1422-0067/24/2/1181 |
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