Rho isoforms have distinct and specific functions in the process of epithelial to mesenchymal transition in renal proximal tubular cells.
Epithelial to mesenchymal transition (EMT) is involved in embryological development, cancerous metastatic spread and organ fibrosis, including the kidney. This process is largely driven by transforming growth factor-beta and recent evidence has implicated Rho as a key intracellular signalling molecule. In this study we have used RNA interference to silence the genetically distinct Rho (A, B and C) isoforms to define their individual functions in human kidney epithelial cells undergoing EMT. We demonstrate that the downregulation of the epithelial cell marker E-cadherin is dependent upon the Rho effector, Rho-kinase. However, silencing RhoA or RhoC expression also results in E-cadherin loss, though each by different mechanisms. Loss of RhoA leads to an upregulation of Snail1 and a reduction in the transcription of E-cadherin whereas loss of RhoC upregulates its breakdown via proteasomal degradation. During EMT, the upregulation of alpha-smooth muscle actin can be blocked by inhibiting the expression of RhoA, but not by that of RhoB or RhoC. This effect is independent of Rho-kinase activity. RhoC is the isoform solely responsible for stress fibre formation and inhibiting its expression reduces EMT-induced migration by 50%. RhoB appears to play a role in cell survival as inhibiting its expression leads to >300% increase in cell apoptosis and a relocalization of focal adhesion kinase. We conclude that Rho is a key signalling molecule in the process of EMT but that each isoform has a distinct and specific role.
Hutchison
Nicol
N
King's College London, Department of Renal Medicine, London, UK.
Hendry
Bruce M
BM
Sharpe
Claire C
CC
D015820:Cadherins / Q000378:metabolism; D002465:Cell Movement; D004847:Epithelial Cells / Q000166:cytology*; D055785:Gene Knockdown Techniques; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000166:cytology*; D008648:Mesoderm / Q000166:cytology*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
D015820:Cadherins; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D053773:Transforming Growth Factor beta1; D020741:rho GTP-Binding Proteins