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The target genes identification of MicroRNA 452 associated with colon cancer and their fuctions
Korea (KMSA) - Wonkwang university, North Jeolla province
Department of pathology
Type of Research Project
- Basic science
What is the background of the project?
MicroRNAs(miRNAs) are non-coding RNAs that play important roles in the pathogenesis of human diseases by regulating target gene expression in specific cells or tissues. We previously identified microRNA452(MIR452) as a colorectal Cancer-associated microRNA using a microarray analysis of colorectal cancer(CRC) tissues and normal colon tissues.
What is the aim of the project?
: We have focused on detecting microRNAs relate to colon cancer, identifying their target molecules, and analyzing the correlation between the microRNAs and their target genes in colon cell lines. Colorectal cancer-associated microRNAs were identified by microRNAs microarray analysis of Colon cancer tissues and normal colon tissues of human. The results were validated by quantitative real time-quantitaive polymerase chain reaction(RT-qPCR). MicroRNA452 target genes were identified by the microRNAs down-regulated in microRNA452-overexpressing cells (determined by microRNA microarray analysis). Luciferase reporter plasmids were constructed to confirm the effect of microRNA452 on target gene expression. The protein expression of the target genes were measured by western blot. Thirty-seven microRNAs were identified as colorectal cancer-associated microRNAs. Colorectal cancer is one of the most common cancers worldwide, and third leading cause of cancer-related mortality in the United States. Its etiology is very complex an not fully understood. Numerous epidemiological and biological studies have suggested various risk factors, including nutrition, physical inactivity, obesity, and diabetes, play a critical role in the etiology of colorectal cancer. Several genetic factors, including microRNAs, are also thought to contribute to colorectal cancer risk. Much evidence suggests that microRNAs are important regulators of oncogenesis. MicroRNAs are enogenously synthesized, short, noncoding RNA molecules of approximately 19-24 nucleotides. MicroRNAs contribute to the post-transcriptional regulation of gene expression in multicellular organisms by controlling the stability and translation of target microRNAs. MicroRNAs contribute to the regulation of crucial biological processes, such as cell proliferation, apoptosis, differentiation, and angiogenesis. They are also implicated in the pathogenesis of various diseases as tumor suppressor genes or oncogenes. Therefore, characterization of microRNA expression patternsin cancer cells may have substantial value for disease diagnosis, prognosis, and therapy.
What techniques and methods are used?
Gene cloning, Luciferase reporter assay. Western blot, quantitative Real time-polymerase chain reaction and cell culture. - Cell culture : The human colorectal cancer cell lines were obtained from Korea Cell Line Bank. - MicroRNA expression profiling : Total RNA(100 ng) is hybridize to an Agilent Human microRNA Microarray (Agilent Technologies, Santa Clara, CA, USA). MicroRNA is labeled, hybridized, and washed following Agilent’s instructions. Images of hybridized microarray images are acquire with a DNA microarray scanner, and the microarray images are analyze with Feature Extraction software. The standard of statistical significance is the correct ratio of the hybridization signal intensity in colon tumor tissue to the hybridization signal intensity in normal tissue. - MicroRNA target prediction by bioinformatics methods : The microRNA targets are predicted using the computer-aided algorithms TargetScan and miRWalk. - Plasmid constructions and luciferase assays. - RNA extraction and quantitative reverse transcription polymerase chain reaction(RT-PCR). - Antibodies and western blot analysis - Statistical analysis
What is the role of the student?
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
We will give students opportunities to experience the basic biomedical science and microRNA functions. Also, students will learn a variety of experimental techniques. They will learn polymerase chain reaction, western-blot & gel electrophoresis that they need to know in the laboratory. They will experience basic social life. They will learn how to operate simple experimental machines. They will learn how to read papers and how to plan experiments.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
No special skills are required
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students
- Khondoker Jahengir Alam; et al. MicroRNA 375 reulates proliferatin ad migration of colon cancr cells by suppressing the CTGF-EGFR signaling pathway. International Journal of Caner. 141; 1614-1629 (2017)
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