*p .05, **p .01. Additionally, a trypan blue cell viability assay was performed to better characterize the effects of miR-96 and miR-183 on cell growth. Data: Quantitation of immunofluorescence data. (XLSX) pone.0233187.s009.xlsx (16K) GUID:?AB1C5452-D328-4459-99E6-AE38877FAAE2 S8 Data: Wound healing images following transfection with miRNA mimics or inhibitors. (PDF) pone.0233187.s010.pdf (576K) GUID:?5DB452C5-B847-4E20-85C6-7B823B90FC99 S9 Data: Immunofluorescence images of vimentin expression in MCF-7M cells. (PDF) pone.0233187.s011.pdf (229K) GUID:?D0CD88D6-FFCA-4FB1-9AFE-DE3C373EDA2F Data Availability StatementAll relevant data are within the manuscript and its Supporting Information files. Abstract Breast cancer is the most commonly diagnosed malignancy in women, and has the second highest mortality rate. Over 90% of all cancer-related deaths are due to metastasis, which is the spread of malignant cells from the primary tumor to a secondary site in the body. It is hypothesized that one cause of metastasis involves epithelial-mesenchymal transition (EMT). When epithelial cells undergo EMT and transition into mesenchymal cells, they display increased levels of cell proliferation and invasion, resulting in a more aggressive phenotype. While many factors regulate EMT, microRNAs have been implicated in driving this process. MicroRNAs are short noncoding RNAs that suppress protein production, therefore loss of microRNAs may promote the overexpression of specific target proteins important for EMT. The goal of this study was to investigate the role of miR-96 and miR-183 in EMT in breast cancer. Both miR-96 and miR-183 were found to be downregulated in post-EMT breast cancer cells. When microRNA mimics were transfected into these cells, there was a significant decrease in cell viability and migration, and a shift from a mesenchymal to an epithelial morphology (mesenchymal-epithelial transition or MET). These MET-related changes may be facilitated in part by the regulation of ZEB1 and vimentin, as both of these proteins were downregulated when miR-96 and miR-183 were overexpressed in post-EMT cells. These findings indicate that the loss of miR-96 and miR-183 may help facilitate EMT and contribute to the maintenance of a mesenchymal phenotype. Understanding the role of microRNAs in regulating EMT is significant in order to not only further elucidate the pathways that facilitate metastasis, but also identify potential therapeutic options for preventing or reversing this process. Introduction Breast cancer is the most commonly diagnosed malignancy in women, with approximately 1 in every 8 women at risk for the disease [1]. There are five clinical subtypes of breast cancer, which are characterized by the nature of the cells that make up the tumor [1]. The most common type of breast cancer, Luminal A, is characterized by an epithelial cell type, which typically indicates a better prognosis due to the low-level of invasiveness of the cells [2]. The characteristics of the epithelial cells found in some breast cancers include tight cell-cell junctions and cell-matrix adhesion, resulting in a cuboidal cell morphology with very low motility [2]. However, other types of breast cancer, such was Basal-like and Claudin-low, display mesenchymal cell characteristics including increased rates of cell growth, invasion, and metastasis [2]. One mechanism that promotes metastasis is the invasion of cancerous cells across the basement membrane, facilitating their entrance into the circulatory or lymphatic system [3]. This can result in the spread of the primary tumor to secondary sites in the body. The metastasis of tumors is responsible for over 90 percent of cancer-related deaths [4], therefore understanding the mechanisms that control this process is crucial to monitoring and treating cancer. It is hypothesized that the first step in the complex metastatic process for carcinomas is epithelial-mesenchymal transition (EMT) [3]. Mesenchymal cells are characterized by their loss of cell-cell junctions and cell-matrix adhesion. Furthermore, during EMT cells undergo changes in cytoskeletal proteins such as the upregulation of vimentin and fibronectin, resulting in a spindle-shaped morphology with increased cellular motility [3]. These changes cause an increase in the invasiveness of the cancer cells. It is hypothesized that EMT is driven by specific molecular changes, including dysregulation.Moreover, there are many other microRNA-based therapeutics currently being developed to treat cancers, with some in clinical trials [19, 20]. data are within the manuscript and its Supporting Information files. Abstract Breast cancer is the most commonly diagnosed malignancy in women, and has the second highest mortality rate. Over 90% of all cancer-related deaths are due to metastasis, which is the spread of malignant cells from the primary tumor to a secondary site in the body. It is hypothesized that one cause of metastasis involves epithelial-mesenchymal transition (EMT). When epithelial cells undergo EMT and transition into mesenchymal cells, they display increased levels of cell proliferation and invasion, resulting in a more aggressive phenotype. While many factors regulate EMT, microRNAs have been implicated in driving this process. MicroRNAs are short noncoding RNAs that suppress protein production, therefore loss of microRNAs may promote the overexpression of specific target proteins important for EMT. The goal of this study was to investigate the role of miR-96 and miR-183 in EMT in breast cancer. Both miR-96 and miR-183 were found to be downregulated in post-EMT breast cancer cells. When microRNA mimics were transfected into these cells, there was a significant decrease in cell viability and migration, and a shift from a mesenchymal to an epithelial morphology (mesenchymal-epithelial transition or MET). These MET-related changes may be facilitated in part by the rules of ZEB1 and vimentin, as both of these proteins were downregulated when miR-96 and miR-183 were overexpressed in post-EMT cells. These findings indicate that the loss of miR-96 and miR-183 may help facilitate EMT and contribute to the maintenance of a mesenchymal phenotype. Understanding the part of microRNAs in regulating EMT is definitely significant in order to not only further elucidate the pathways that PP1 facilitate metastasis, but also determine potential therapeutic options for avoiding or reversing this process. Introduction Breast tumor is the most commonly diagnosed malignancy in ladies, with approximately 1 in every 8 ladies at risk for the disease [1]. You will find five medical subtypes of breast cancer, which are characterized by the nature of the cells that make up the tumor [1]. The most common type of breast tumor, Luminal A, is definitely characterized by an epithelial cell type, which typically shows a better prognosis due to the low-level of invasiveness of the cells [2]. The characteristics of the epithelial cells found in some breast cancers include limited cell-cell junctions and cell-matrix adhesion, resulting in a cuboidal PP1 cell morphology with very low motility [2]. However, other types of breast tumor, such was Basal-like and Claudin-low, display mesenchymal cell characteristics including increased rates of PP1 cell growth, invasion, and metastasis [2]. One mechanism that promotes metastasis is the invasion of cancerous cells across the basement membrane, facilitating their entrance into the circulatory or lymphatic system [3]. This can result in the spread of the primary tumor to secondary sites in the body. The metastasis of tumors is responsible for over 90 HES7 percent of cancer-related deaths [4], consequently understanding the mechanisms that control this process is vital to monitoring and treating cancer. It is hypothesized the first step in the complex metastatic process for carcinomas is definitely epithelial-mesenchymal transition (EMT) [3]. Mesenchymal cells are characterized by their loss of cell-cell junctions and cell-matrix adhesion. Furthermore, during EMT cells undergo changes in cytoskeletal proteins such as the upregulation of vimentin and fibronectin, resulting in a spindle-shaped morphology with increased cellular motility [3]. These changes cause an increase in the invasiveness of the tumor cells. It.