Dephosphorylation of the Retinoblastoma protein (Rb) inhibits cancer cell EMT via Zeb.
The tumor suppressor Retinoblastoma (Rb) protein is highly phosphorylated in cancer cells largely due to the overexpression of cyclins or the loss of expression of cyclin dependent kinase inhibitors (cdki). Hyperphosphorylation of Rb promotes proliferation, and plays a role in the regulation of apoptosis. Recently, inhibition of cyclin dependent activity toward Rb has been identified as a strategy that has shown clinical efficacy. We utilized a method to induce phosphatase activity toward Rb in cells by shRNA silencing of PNUTS (Phosphatase Nuclear Targeting Subunit) that regulates PP1-mediated dephosphorylation of Rb. In this study, the effect of Rb dephosphorylation on the epithelial to mesenchymal transition (EMT) was determined. The EMT transition is observed in cancer cells that have acquired invasive characteristics. In breast cancer cells grown in 3D Matrigel cultures, MCF7 cells undergo apoptosis in response to Rb dephosphorylation, whereas MDA-MB-231 and Hs578T cells exhibit a reduction in the EMT. Cells devoid of phosphorylated Rb (nontransformed MCF10A and Rb-null MDA-MB-468) lacked any response to PNUTS depletion, showing the effect is Rb-dependent. In addition, these studies showed that Rb dephosphorylation in 3D Matrigel cultures of highly invasive HT1080 cells led to the inhibition of the EMT. Furthermore we observed association between dephosphorylated Rb with ZEB1, a zinc-finger E-box-binding transcription factor that regulates expression of E- and N-cadherins. Finally Rb dephosphorylation led to inhibition of ZEB1 transcriptional activity, this data supports the notion that Rb dephosphorylation modulates the EMT. These studies suggest targeting Rb phosphorylation in mesenchymal cancer cells may decrease invasiveness.
['Egger|Jacklynn V|JV|', 'Lane|Maria V|MV|', 'Antonucci|Lisa A|LA|', 'Dedi|Brixhilda|B|', 'Krucher|Nancy A|NA|']
D017209:Apoptosis D001943:Breast Neoplasms D045744:Cell Line, Tumor D049109:Cell Proliferation D058750:Epithelial-Mesenchymal Transition D005260:Female D006801:Humans D061986:MCF-7 Cells D010766:Phosphorylation D016160:Retinoblastoma Protein D014162:Transfection D000071799:Zinc Finger E-box-Binding Homeobox 1
Retinoblastoma EMT Apoptosis phosphatase MCF7 ZEB1 TNBC