P-215: Discovery of A Novel APA Variant of A Human Potential Gene Based on Expressed Sequenced Tags Analysis

Background: Expressed sequence tags (ESTs) are sequences of cDNA fragments prepared from different tissue sources. There are over one million of these sequences in the publicly available database, and these sequences are believed to represent more than half of all human genes. The ESTs belong to different cDNA libraries, was prepared from one particular cell type, organ, or tumor. Therefore, the presence or absence of ESTs in different libraries provides information about the organ, cell type, or tumor specificity of expressed genes. Also, a gene is often represented by many ESTs; generally, the more a gene is expressed in a given tissue, the more ESTs for that gene will be found in the library. Materials and Methods: 1. Computational study: Bioinformatics Analysis showed some important ESTs can be representation of a new potential gene. Important ESTs were selected to design specific primers. 2. Cell Culturing: U87-MG cell line cultured in RPMI medium. RNA was extracted and then single strand cDNA was prepared based on oligo dT primer. RT-PCR was performed by nested primers for our based EST and then its sequence confirmed by DNA sequencing. Results: We have discovered a novel alternative polyadenylation variant that encoded by our potential gene, experimentally. This variant has a potential role in tumorigenesis. Our sequencing data revealed this variant can misses a spicific region that potentially has a binding site for a known micro-RNA. Conclusion: Based on bioinformatics approaches, our study revealed this novel variant that is longer than our based EST has a target site for has-miR-130a-5p but it needs an experimental validation. Expression of this variant in U87-MG cell line but not in normal tissues, supports this hypothesis that 3’UTR of mRNA involves in tumorigenesis.