RHBDD2 AND WWOX PROTEIN INTERACTION AS MODULATOR MECHANISM OF PROLIFERATION AND DIFFERENTIATION OF NORMAL AND NEOPLASTIC BREAST CELLS

FERRETTI V; CANZONERI R; LACUNZA E; KORDON E; ALDAZ CM; ABBA MC

Congreso; LXI REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE INVESTIGACIÓN CLÍNICA; 2016

The 􀀴homboid gene family constitutes a heterogeneous group of proteases / pseudoproteases that are conserved throughout evolution, involved in different biological processes such as EGFR, TNF, ERAD and UPR signaling pathways among others. 􀀹e previously demonstrated that RHBDD2 is a cancer relatedgene amplified and overexpressed in advanced breast cancer. On the other and, The WW domain containing protein WWOX has been postulated as a tumor suppressor in breast and other cancers. WWOX modulates the TGFB signaling path􀁙ay via direct WW-domain mediated binding and potential cytoplasmic sequestration of S􀀯A􀀦􀀕 protein. The aim of this study 􀁙as to characterize the RHBDD2 protein interaction and theirbiological relevance over the TGF􀁅 signaling path􀁙ay in normaland breast cancer cells. Functional proteomics analysis using TA􀀲/􀀯S allo􀁙ed us to identify a 􀀴H􀀤􀀦􀀦􀀔 􀅌 􀀹􀀹􀀱􀀺 protein􀀏proteininteraction in HE􀀭􀀔􀀛􀀕 cancer cells. This result 􀁙as validated innormal 􀀊HC􀀓􀀓􀀋 and breast cancer cell lines 􀀊􀀯CF􀀙, T􀀖􀀙􀀦, 􀀯􀀦-A􀀯􀀤􀀔􀀕􀀓􀀋 by Co􀀏immunoprecipitation 􀀊Co􀀏􀀫􀀲􀀋 and 􀁙estern􀀏blot. 􀀫naddition, co􀀏localization analysis corroborate the previous finding,demonstrating that such protein-protein interaction occurs at the􀀩olgi complex. HC􀀓􀀓 mouse mammary cell line 􀁙as employedto evaluate 􀀴hbdd􀀔 and 􀀹􀁙ox expression at m􀀴􀀰A and proteinlevels by 􀀴T􀀏􀀲C􀀴 and Co􀀏􀀫􀀲/􀀹􀀤 methods respectively. 􀀹edetected a high 􀀴hbdd􀀔 expression at the proliferative stage ofthe HC􀀓􀀓 cells 􀁙ith a dramatic decrease at the competent anddifferentiated stages, 􀁙hile 􀀹􀁙ox expression 􀁙as expressed atthe differentiated stage. 􀀱ligo􀀏microarray based analysis of 􀀯􀀦-􀀯􀀤A􀀔􀀕􀀓 􀀴H􀀤􀀦􀀦􀀔 silenced cells 􀀊si􀀴􀀰A􀀋 suggest that 􀀴H􀀤􀀦􀀦􀀔could be involved in the modulation of T􀀩F􀁅 signalling path􀁙ayamong other bioprocess via sequestration of 􀀹􀀹􀀱􀀺 protein atthe 􀀩olgi complex. This study sho􀁙 that the 􀀴H􀀤􀀦􀀦􀀔􀀏􀀹􀀹􀀱􀀺protein interaction is associated 􀁙ith the modulation of the mammary proliferation and differentiation process, suggesting that thisinteraction might play a relevant role in the normal developmentand neoplastic progression