Detected the Expression of Large Non-coding RNA on Hepatocellular Carcinoma (HCC) using the FISH Method

• Main Authors: Li-Li Lin, 林莉莉
• Other Authors: Jan-Gowth Chang
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/74921184242434770411

碩士 === 高雄醫學大學 === 醫學研究所 === 98 === In human genome，only about 2% of nucleotides can encode proteins，while over 90% of the genome have non-protein coding transcripts (called non-coding RNA)。 In spite of much study focusing on miRNA, rRNA and tRNA，most of ncRNA have rarely been studied。 The cancer-related studies of ncRNA，excluding miRNA and a minor part of ncRNA，are also rarely studied， especially for the role of the very long ncRNA (over 5kb) in the development of cancers。The aim of this study is to use fluorescence in situ hybridization(FISH) method to analyze and compare the expression of ncRNA between Hepatocellular carcinoma (HCC) cancer and non-cancer cells。 In this experiment，the bioinformatic method was used to get 40 large ncRNA（over 5kb）. Using Real-time quantitative polymerase chain reaction (Real-time quantitative PCR) to analyze the expression of these large ncRNA in HCC. The result showed that the expression of HCC cancer cells was different from that of their neighbor normal cells。 The results were further confirmed using formalin fixed tissue and stain with fluorescence in situ hybridization(FISH)。 We therefore can use the paraffin section obtained from the HCC patient to observe the expression of large ncRNA in the liver tissue。Not only this method can use many years old the paraffin block , also it can observe large ncRNA in the cell''s distribution, and it can not be achieved by Real-time quantitative PCR .