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Title:
Rab31-dependent regulation of transforming growth factor ß expression in breast cancer cells | Molecular Medicine
Description:
Background The small GTP-binding protein Rab31 plays an important role in the modulation of tumor biological-relevant processes, including cell proliferation, adhesion, and invasion. As an underlying mechanism, Rab31 is presumed to act as a molecular switch between a more proliferative and an invasive phenotype. This prompted us to analyze whether Rab31 overexpression in breast cancer cells affects expression of genes involved in epithelial-to-mesenchymal transition (EMT)-like processes when compared to Rab31 low-expressing cells. Methods Commercially available profiler PCR arrays were applied to search for differentially expressed genes in Rab31 high- and low-expressing CAMA-1 breast cancer cells. Differential expression of selected candidate genes in response to Rab31 overexpression in CAMA-1 cells was validated by independent qPCR and protein assays. Results Gene expression profiling of key genes involved in EMT, or its reciprocal process MET, identified 9 genes being significantly up- or down-regulated in Rab31 overexpressing CAMA-1 cells, with the strongest effects seen for TGFB1, encoding TGF-ß1 (> 25-fold down-regulation in Rab31 overexpressing cells). Subsequent validation analyses by qPCR revealed a strong down-regulation of TGFB1 mRNA levels in response to increased Rab31 expression not only in CAMA-1 cells, but also in another breast cancer cell line, MDA-MB-231. Using ELISA and Western blot analysis, a considerable reduction of both intracellular and secreted TGF-ß1 antigen levels was determined in Rab31 overexpressing cells compared to vector control cells. Furthermore, reduced TGF-ß activity was observed upon Rab31 overexpression in CAMA-1 cells using a sensitive TGF-ß bioassay. Finally, the relationship between Rab31 expression and the TGF-ß axis was analyzed by another profiler PCR array focusing on genes involved in TGF-ß signaling. We found 12 out of 84 mRNAs significantly reduced and 7 mRNAs significantly increased upon Rab31 overexpression. Conclusions Our results demonstrate that Rab31 is a potent modulator of the expression of TGF-ß and other components of the TGF-ß signaling pathway in breast cancer cells.
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Keywords {🔍}
rab, cells, cell, cancer, pubmed, tgfß, expression, article, google, scholar, levels, breast, cas, genes, cama, control, protein, overexpressing, signaling, vector, tumor, overexpression, proliferation, culture, pcr, tgfb, fig, growth, germany, central, profiler, emt, supernatants, analysis, pathway, mrna, array, elisa, min, tgfβ, biol, mdamb, human, total, processes, compared, identified, downregulation, proteins, lysates,
Topics {✒️}
akt/gsk-3β/snail signaling akt/gsk-3β/snail pathway small gtp-binding proteins growth factor-binding sequence permanox-type lab-tek slides tgf-β/bone morphogenic protein rabbit anti-goat igg tgf-ß/activin signaling cascades tgf-ß-induced actin remodeling measuring bioactive tgf-beta tgf-ß1/bmp array measure active tgf-ß rho/rac family tgfβ-induced transcription tgf-β1 induces emt mda-mb-231 rab31-overexpressing cells fluorochrome-coupled polyclonal antibody facilitates tgf-β signaling active tgf-ß levels mir-30c-2-3p regulates detect active tgf-ß1 tgf-ß-deficient mice article download pdf tgf-β1 antigen levels polyclonal antibody rt3-igg tgf-β1 protein expression tgf-ß reporter plasmid tgf-ß signaling pathway human tgf-ß levels tgf-β-induced emt tgf-ß induces emt mann–whitney u-test tgf-ß signaling axis universal probe-libray probes tgf-ß-induced emt tgfbeta signalling activate latent tgf-ß1 total tgf-ß1 levels tgf-ß1 mrna levels tgf-ß1 specific antibody ccr7/ccl21-mediated chemotaxis rab-gtpase regulates emt tgf-ß expression levels dose-dependent manner total tgf-ß content tgf-beta signaling profiler pcr array tgf-ß1 mrna expression rab31 low-expressing cells sensitive tgf-ß bioassay
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headline:Rab31-dependent regulation of transforming growth factor ß expression in breast cancer cells
description:The small GTP-binding protein Rab31 plays an important role in the modulation of tumor biological-relevant processes, including cell proliferation, adhesion, and invasion. As an underlying mechanism, Rab31 is presumed to act as a molecular switch between a more proliferative and an invasive phenotype. This prompted us to analyze whether Rab31 overexpression in breast cancer cells affects expression of genes involved in epithelial-to-mesenchymal transition (EMT)-like processes when compared to Rab31 low-expressing cells. Commercially available profiler PCR arrays were applied to search for differentially expressed genes in Rab31 high- and low-expressing CAMA-1 breast cancer cells. Differential expression of selected candidate genes in response to Rab31 overexpression in CAMA-1 cells was validated by independent qPCR and protein assays. Gene expression profiling of key genes involved in EMT, or its reciprocal process MET, identified 9 genes being significantly up- or down-regulated in Rab31 overexpressing CAMA-1 cells, with the strongest effects seen for TGFB1, encoding TGF-ß1 (> 25-fold down-regulation in Rab31 overexpressing cells). Subsequent validation analyses by qPCR revealed a strong down-regulation of TGFB1 mRNA levels in response to increased Rab31 expression not only in CAMA-1 cells, but also in another breast cancer cell line, MDA-MB-231. Using ELISA and Western blot analysis, a considerable reduction of both intracellular and secreted TGF-ß1 antigen levels was determined in Rab31 overexpressing cells compared to vector control cells. Furthermore, reduced TGF-ß activity was observed upon Rab31 overexpression in CAMA-1 cells using a sensitive TGF-ß bioassay. Finally, the relationship between Rab31 expression and the TGF-ß axis was analyzed by another profiler PCR array focusing on genes involved in TGF-ß signaling. We found 12 out of 84 mRNAs significantly reduced and 7 mRNAs significantly increased upon Rab31 overexpression. Our results demonstrate that Rab31 is a potent modulator of the expression of TGF-ß and other components of the TGF-ß signaling pathway in breast cancer cells.
datePublished:2021-12-14T00:00:00Z
dateModified:2021-12-14T00:00:00Z
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TGF-ß signaling
GTPase
Rab31
Breast cancer
EMT
Molecular Medicine
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headline:Rab31-dependent regulation of transforming growth factor ß expression in breast cancer cells
description:The small GTP-binding protein Rab31 plays an important role in the modulation of tumor biological-relevant processes, including cell proliferation, adhesion, and invasion. As an underlying mechanism, Rab31 is presumed to act as a molecular switch between a more proliferative and an invasive phenotype. This prompted us to analyze whether Rab31 overexpression in breast cancer cells affects expression of genes involved in epithelial-to-mesenchymal transition (EMT)-like processes when compared to Rab31 low-expressing cells. Commercially available profiler PCR arrays were applied to search for differentially expressed genes in Rab31 high- and low-expressing CAMA-1 breast cancer cells. Differential expression of selected candidate genes in response to Rab31 overexpression in CAMA-1 cells was validated by independent qPCR and protein assays. Gene expression profiling of key genes involved in EMT, or its reciprocal process MET, identified 9 genes being significantly up- or down-regulated in Rab31 overexpressing CAMA-1 cells, with the strongest effects seen for TGFB1, encoding TGF-ß1 (> 25-fold down-regulation in Rab31 overexpressing cells). Subsequent validation analyses by qPCR revealed a strong down-regulation of TGFB1 mRNA levels in response to increased Rab31 expression not only in CAMA-1 cells, but also in another breast cancer cell line, MDA-MB-231. Using ELISA and Western blot analysis, a considerable reduction of both intracellular and secreted TGF-ß1 antigen levels was determined in Rab31 overexpressing cells compared to vector control cells. Furthermore, reduced TGF-ß activity was observed upon Rab31 overexpression in CAMA-1 cells using a sensitive TGF-ß bioassay. Finally, the relationship between Rab31 expression and the TGF-ß axis was analyzed by another profiler PCR array focusing on genes involved in TGF-ß signaling. We found 12 out of 84 mRNAs significantly reduced and 7 mRNAs significantly increased upon Rab31 overexpression. Our results demonstrate that Rab31 is a potent modulator of the expression of TGF-ß and other components of the TGF-ß signaling pathway in breast cancer cells.
datePublished:2021-12-14T00:00:00Z
dateModified:2021-12-14T00:00:00Z
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TGF-ß signaling
GTPase
Rab31
Breast cancer
EMT
Molecular Medicine
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Analytics and Tracking {📊}
- Google Tag Manager
Libraries {📚}
- Clipboard.js
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CDN Services {📦}
- Crossref