ARRA: Alcohol and Breast Cancer

The goal of parental proposal is to elucidate the cellular/molecular mechanism underling alcohol induced promotion of human mammary tumors. Our central hypothesis was that ethanol induced oxidative stress upregulates MUC1 as an adaptor protein to promote ErbB2/ƒÀ]catenin interaction which induces dissociation of the ƒÀ]catenin/E]cadherin complex, leading to cell transformation and cell migration/invasion. In the parental proposal, we planned to investigate the effect of ethanol on the interaction among ErbB2, MUC1, E cadherin and ƒÀ catenin using biochemical approaches, such as co immunoprecipitation or protein binding assays. In the supplement, we will directly examine the protein/protein interaction and their redistribution using a time]lapse live cell image system and immunofluorescent microscopy. With this approach, we will investigate protein association/dissociation and redistribution of these complexes in different intracellular compartments, such as cytoplamic membrane, endoplasmic reticulum or nucleus. The new approaches will offer several advantages; these include 1.. we can directly visualize the distribution and co-localization of ErbB2, MUC1, E Cadherin, and ƒÀ catenin in different intracellular compartments; 2.. we are able to precisely monitor the time sequential effect of ethanol on the protein/protein interaction and their redistribution using time lapse live cell image; 3.. we are able to precisely monitor ethanol induced intracellular ROS production using immunofluorescent probes and correlate the changes in ROS profile with alterations in protein/protein interaction and redistribution. The new approach will provide more direct, accurate and sensitive detection of ethanol's effect on protein/protein interaction, distribution and the consequence of these interactions. The study will provide critical insight into the cellular/molecular mechanisms underlying alcohol induced tumor promotion and offer potential preventive/therapeutic strategy for treating breast cancers. Thus, the supplement will not only significantly enhance the scope and impact of parental proposal, but also stimulating the economy by procuring additional equipment and recruiting for additional needed skills.