| Summary: | 碩士 === 國立臺灣大學 === 生物科技研究所 === 105 === MicroRNAs (miRNAs) are endogenous non-coding RNAs of 22-24 nucleotides in length that regulate gene expression by complementary binding to the 3’ untranslated regions (3’UTR) of specific mRNA targets. In recent years, accumulating evidences suggested that dysregulated miRNAs can function either as tumor suppressor or oncogenes to affect progression, proliferation, metastasis and drug sensitivity of tumor cells. In addition, many studies mentioned that miRNAs were associated with patients’ prognosis, overall survival and recurrence and may play important roles in tumor progression and prognosis. Our previous study showed that the miR-338-5p expression level was higher in the non-recurrence esophageal squamous cell carcinoma (ESCC) patient samples than that in the recurrence ESCC patient samples by next generation sequencing (NGS) analysis. According to this result, miR-338-5p was considered a miRNA that may relate to ESCC patients’ recurrence. Therefore, the aim of this study is to study the possible roles of miR-338-5p and its underlying mechanisms in ESCC cells. We first transfected miR-338-5p mimic into CE81T/VGH cells, which is an ESCC cell line with higher migration, proliferation and survival ability than other ESCC cell lines. The results showed that the cell migration, proliferation and survival were significantly reduced in the cells with mimic-miR-338-5p using transwell assay, MTT assay and colony formation assay. In order to better understand regulatory mechanisms of miR-338-5p in ESCC cells, miRSystem and Ingenuity Pathways Analysis were used to predict the target genes of miR-338-5p. FERMT2 was identified and validated as the target genes of miR-338-5p using luciferase reporter assay. Furthermore, the results showed that the cell migration, proliferation and survival were significantly reduced in the cells with lower FERMT2 expression using transwell assay, MTT assay and colony formation assay in ESCC cell line. Next, we detected the cisplatin induced cytotoxicity in the CE81T/VGH cells and the results demonstrated that the cells with miR-338-5p overexpression or reduced FERMT2 expression could increase cisplatin induced cytotoxicity in the CE81T/VGH cells. Moreover, we analyzed the relationships between FERMT2 expression levels and the survival time of ESCC patients, and the result indicated that FERMT2 could be a good biomarker to predict the survival outcome of esophageal cancer patients. In summary, our study suggested that miR-338-5p might reduce cell proliferation, migration, survival, and to enhance the cisplatin sensitivity in the ESCC cells through FERMT2 regulation.
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