Harnessing 3D models of mammary epithelial morphogenesis: An off the beaten path approach to identify candidate biomarkers of early stage breast cancer. Issue 2 (1st October 2016)

• Record Type: Journal Article
• Authors: Rossetti, Stefano; Bshara, Wiam; Reiners, Johanna A.; Corlazzoli, Francesca; Miller, Austin; Sacchi, Nicoletta
• Is Part Of: Cancer letters. Volume 380:Issue 2(2016)
• Page Start: 375
• Page End: 383
• Publication Date: 2016-10-01
• Subjects: 3D mammary epithelial morphogenesis -- Breast cancer -- Ductal carcinoma in situ (DCIS) -- Biomarkers -- Annexins; 2D two dimensional -- 2D DIGE two dimensional difference gel electrophoresis -- 3′ UTR 3′ untranslated region -- 3D three dimensional -- ADH atypical ductal hyperplasia -- ANXA2 annexin A2 -- ANXA8 annexin A8 -- C.I. confidence interval -- DCIS ductal carcinoma in situ -- ERA estrogen receptor alpha -- EV empty vector -- HME1 human mammary epithelial cell line -- HME1-MYC HME1 cells overexpressing MYC -- HME1-shERA ERA knock down HME1 cells -- HME1-shMTG16 MTG16 knock down HME1 cells -- HME1-shPER2 PER2 knock down HME1 cells -- HME1-shANXA2 ANXA2 knock down cells -- IPA ingenuity pathway analysis -- miRNA microRNA -- MW molecular weight -- PI isoelectric point -- PER2 Period 2 -- RA retinoic acid -- RARA403 dominant negative RA receptor alpha -- SCR scrambled -- TMA tissue microarray; Cancer -- Periodicals; Neoplasms -- Periodicals; Cancer -- Périodiques; Electronic journals; 616.994
• DOI: 10.1016/j.canlet.2016.07.003
• Languages: English
• ISSNs: 0304-3835

Abstract

Highlights: HME1 cells with distinct genetic mutations develop similar 3D DCIS-like morphology. HME1 cells developing a 3D DCIS-like morphology share a core protein signature. Inverse deregulation of ANXA2 and ANXA8 is a feature of DCIS-like HME1 precursors. ANXA8 expression is significantly higher in DCIS vs normal breast tissue and ADH. Abstract: Regardless of the etiological factor, an aberrant morphology is the common hallmark of ductal carcinoma in situ (DCIS), which is a highly heterogeneous disease. To test if critical core morphogenetic mechanisms are compromised by different mutations, we performed proteomics analysis of five mammary epithelial HME1 mutant lines that develop a DCIS-like morphology in three dimensional (3D) culture. Here we show first, that all HME1 mutant lines share a common protein signature highlighting an inverse deregulation of two annexins, ANXA2 and ANXA8. Either ANXA2 downregulation or ANXA8 upregulation in the HME1 cell context are per se sufficient to confer a 3D DCIS-like morphology. Seemingly, different mutations impinged on a common mechanism that differentially regulates the two annexins. Second, we show that ANXA8 expression is significantly higher in DCIS tissue samples versus normal breast tissue and atypical ductal hyperplasia (ADH). Apparently, ANXA8 expression is significantly more upregulated in ER-negative versus ER-positive cases, and significantly correlates with tumor stage, grade and positive lymph node. Based on our study, 3D mammary morphogenesis models can be an alternate/complementary strategy for unraveling new DCIS mechanisms and biomarkers.