| Summary: | 碩士 === 長庚大學 === 生物醫學研究所 === 97 === MicroRNAs (miRNAs) are short 20~22 nucleotide RNA molecules that function as negative regulators of gene. Numerous studies have found that miRNAs are aberrantly expressed in cancer and demonstrated that some miRNAs may function as oncogenes or tumor suppressor genes.
For the purpose of understanding the roles of miRNA in the pathogenesis of glioblastoma multiform (GBM), we used a quantitative reverse transcription polymerase chain reaction (qRT-PCR) to profile 29 GBM tissue samples and found that human microRNA miR-196b is significantly elevated in GBM samples. We also measured the expression level of miR-196b in brain tumor cell lines derived from high grade GBM that have higher level of miR-196b. Base on our finding, we suspect that miR-196b may participate in the tumorgenesis of brain tumor.
In order to explore the function of miR-196b in GBM, we constructed Lentiviral vectors and expressed miR-196b in cell lines. Over-expression of mir-196b in brain cell line leads to promote cell growth, survival and soft agar anchorage-independent growth. Phalanx gene expression array was used to analyze mRNA expression and more than 764 genes were found to be differential expression upon miR-196b overexpression. Computational algorithms have played a central role in the target prediction for microRNAs. Using an in-house miRNA target prediction script, we identified 897 high efficacy human targets. Then we used statistical enrichment analysis to identify Gene Ontology categories as well as biological functions and signaling pathways regulated by miR-196b. In addition, we combined both results and identified 9 genes that could be potential direct targets of miR-196b.
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