| Summary: | 碩士 === 國立陽明大學 === 口腔生物研究所 === 107 === Head and neck carcinoma is among the top five cancer deaths in males of Taiwan. To understand the pathogenesis of oral squamous cell carcinoma (OSCC) and find either biomarkers or therapeutic targets is important. MicroRNA (miRNA) are short non-coding RNA and play important roles in the regulation of gene expression and biological development. miRNAs deregulated cancer development and progression. Deregulated expression of miRNAs is involved in tumor progression. Recent studies have shown that miR-371/372/373 cluster, localized in human chromosome 19q13.4, is co-expressed in human embryonic stem cells and acts as an oncogene in several cancers including hepatoblastoma, parathyroid carcinomas, colorectal cancer and others. The miR-371/372/373 has been found to enhance cancer formation and progression. Our previous studies have evidenced that the individual miR-371/372/373 is highly expressed in OSCC. However, the role of miR-371/372/373 cluster in oral carcinogenesis remains obscure. We use CRISPR-Cas9 system to establish the cell subclones having the deletion of miR-371/372/373 cluster. Cell subclones exhibiting the depletion of the promoter of this gene cluster are also established. CRISPR-dcas9 SAM system are used to induce endogenous miR-371/372/373 expression by promoter activation. The analysis of miR-371/372/373 cluster deletion and promoter deletion cell subclones shows a decrease in oncogenic phenotypes. In addition, deletion of miR-371/372/373 gene cluster or its promoter increases the sensitivity of the cisplatin. DKK1 is amongst the known targets most eminently up-regulated in miR-371/372/373 deletion subclones. This study also shows that the depletion of the promoter is unable to affect the expression of genes localized juxtaproximal to miR-371/372/373 cluster. Conversely, the activation of the miR-371/372/373 promoter enriches the oncogenic phenotypes of SAS line and activates FAK, AKT, ERK and Src signaling elements which are associated with cancer progression.
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