Elucidating the Role of miR-148a in Gastric Cancer AGS Cells Using RNA-seq

• Main Authors: Kuan-Hao Hsu, 徐寬豪
• Other Authors: Hsueh-Fen Juan
• Format: Others
• Language: en_US
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/25760812776815055507

碩士 === 國立臺灣大學 === 分子與細胞生物學研究所 === 101 === MicroRNAs play an important role in various biological processes by post-transcriptionally regulating gene expression. To investigate the role of miR-148a in gastric cancer, the next generation RNA sequencing (RNA-seq) was applied to reveal the miR-148a-regulated gene expression profiles in gastric cancer cells. To uncover the role of miR-148a in gastric cancer, miR-148a was overexpressed in gastric cancer cell line AGS and followed by RNA-seq analysis. Using RNA-seq analysis, gene expression levels were estimated to identify differentially expressed genes in miR-148a-overexpressed cells. The biological processes and functional networks associated with the regulated genes were further analyzed by Ingenuity Pathway analysis (IPA). To validate the finding from RNA-seq, the expression level of the interested genes was measured by real-time reverse transcription PCR (qRT-PCR), the cell growth was monitored in real-time using the xCELLigence system, and the cell cycle analysis was examined by flow cytometry. We identified a total of 16,254 genes by comparing RNA-seq profiles of negative control (NTC) and miR-148a-overexpressed (Pre) AGS cells. Among them, 63 up-regulated and 48 down-regulated genes were differentially expressed in miR-148a-overexpressed cells. The IPA results showed that these miR-148a-regulated genes were significantly involved in the biological pathways including cell death and cell cycle. We demonstrated overexpression of miR-148a reduced the cell growth and alter the cell cycle G2/M phase population increased. This study illustrates the new role of miR-148a in controlling cell cycle by RNA-seq and provides information in gastric cancer therapy.