Lnc-EPCAM AND Lnc-BHLHE41 AS RNA REGULATORS OF BREAST CANCER AND BREAST CANCER PREVENTION

Abstract

The objective of this study was to unveil a novel area of gene regulation in breast cancer and breast cancer prevention through the study of a recent discovered class of genetic regulators named long non-coding RNAs (lncRNAs). LncRNAs are RNA molecules longer than 200 nucleotides that are not translated into proteins, but regulate the transcription of genes involved in different cellular processes, including differentiation, cancer initiation and progression. The link between lncRNAs and cancer is well documented in the literature. More recently, their relevance in the transcription field is beginning to be explored and their roles have been found to vary from guiding proteins to the genome to scaffolding proteins complexes needed for the transcription of a specific gene. Initial transcriptome analysis of normal breast of parous and nulliparous postmenopausal women revealed that several lncRNAs are differentially expressed in the parous breast. This observation provided evidence of a potential role of lncRNAs in the regulation of transcription and their function in pregnancy’s preventive effect in reducing the lifetime risk of developing breast cancer. Specifically, RNA sequencing of healthy postmenopausal breast tissue biopsies from eight parous and eight nulliparous women using Illumina platform was performed. The sequencing results showed that there are 42 lncRNAs differentially expressed between parous and nulliparous breast tissue. These data led to the hypothesis that these novel lncRNAs may be drivers in the process of development that occurs in the mammary gland during pregnancy, providing protection against breast cancer. After analysis of these 42 lncRNAs using bioinformatics tools, review of the scientific literature, and real-time PCR analysis, two lncRNAs (lncBHLHE41 and lncEPCAM) were selected to be tested in vitro, using different molecular techniques in human epithelial breast cell lines to determine their relevance in breast cancer. This project provided novel information on lncRNAs induced by pregnancy in the breast tissue, and identified two lncRNAs as potential key regulators in breast differentiation and cancer progression. The manipulation of these lncRNAs led to evidence of their function in vitro and, using xenograft studies, we determined their relevance in vivo. Although treatment for cancer using lncRNAs as targets is in its infancy at the clinic, the advancement in knowledge and technology to study their relevance in disease could lead to the development of therapeutics for breast cancer and breast cancer prevention in the near future.