Expression and functional analysis of LINC۰۰۵۱۳ in colorectal carcinoma tissue and cell lines and its association with Wnt signaling.

Background and Aim: The colorectal cancer is one of the most frequent malignant diseases worldwide with a remarkably high mortality rate. The cancer involves a slowly developing cancer that begins as a tumor or tissue growth on the rectum or colon. The main causes of the cancer include lifestyle, personal characteristics, and inheritance. The symptoms of colon cancer include: a persistent change in your bowel habits, rectal bleeding or blood in your stool, weakness or fatigue, and unexplained weight loss. The Wnt signaling pathway is important for both initiation and progression of CRC. The Wnt target genes affect multiple cellular functions including regulators of cell cycle progression, cell proliferation, angiogenesis, and apoptosis. A key initial mutation is the early mutation of the adenomatous polyposis coli (APC) tumor suppressor gene. The function of this tumor suppressor is to bind with the b-catenin and induces its degradation, thereby acting as a negative regulator of b-catenin. The Long non-coding RNAs (lncRNAs) are members of the non-protein coding RNA family. They play a critical role in the regulation of gene and protein expression influencing apoptosis, cell proliferation and they have a critical role in the development and progression of various cancers, including colorectal cancer (CRC). MicroRNAs (miRNAs) are important regulatory molecules which could affect a variety of cellular and molecular targets in CRC.Methods: ۱.Sample preparation and RNA extraction from different colorectal carcinoma specimens ۲.cDNA synthesis and qPCR analysis of the candidate LncRNA and Data interpretation ۳.Cloning of the candidate LncRNA in an expression vector ۴.Overexpression of the cloned LncRNA in colorectal cell lines (SW۴۸۰, HCT۱۱۶) ۵.Assessment of LncRNA expression on cell cycle and cell death ۶.Study of the LncRNA expression change on the target genes of the interacting miRNA ۷.Cloning of the LncRNA downstream of a luciferase ORF for luciferase reporter assay ۸.Study of the interaction between the candidate LncRNA and the corresponding miRNAResults: We aim to find a LncRNA from multiple bioinformatics databases and its effect on CRC. We found Linc۰۰۵۱۳ and investigate the expression level for this LncRNA and its correlation with candidate MiRNAs from normal and abnormal colorectal tissues. We report a significant difference in expression level of this gene on normal and abnormal tissue and start experimental work on colorectal cancer and normal tissue.Conclusion: We investigate the preclinical and clinical studies related to miRNAs and LncRNAs as diagnostic biomarkers for CRC.

Keywords:

Long non-coding RNA , Colorectal cancer , Micro RNA