Study of the membrane dot blot on K-ras oncogen detection

• Main Author: 翁麗卿
• Other Authors: 曾博修
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/70794794439049321397

碩士 === 中山醫學大學 === 生化暨生物科技研究所 === 98 === K-ras oncogen is frequently found in human cancer. Activating K-ras mutations are present in colorectal adenomas carcinoma、 lung cancer、 breast cancer and pancreatic tumor. Several therapeutic agents have been developed in order to inhibit K-ras activation. However, there is no diagnostic tool available to detect oncogenic K-ras activation in patient，s specimens so far. Therefore, researchers have been developed a new K-ras diagnostic membrane arrays in 2004, trying to resolve the problem. It may monitor and control cancer transformation and malignant progression. In the past, the detection of K-ras mutation was mainly relying on direct sequencing, however the sequencing method could not provide researchers information but mutation sites. Using the new K-ras membrane array can detect circulating cancer cells with mutant K-ras oncogen in peripheral blood. This is a low-cost, convenient, and easy technique for detecting activated K-ras in a small number of circulating tumor cells by the colorimetric membrane array method (CLMA); but the sensitivity still needs improvement. The object of this study is to develop a new platform with chemiluminescence as reporter and weighted values of target genes on the chip in order to achieve a more sensitive, easier to read, and more accurate platform-membrane dot blot on K-ras oncogen detection . In advance, we collected cancer tissues from 220 cancer patients including breast cancer, colorectal cancer, and NSCLC cancer to evaluate the membrane dot blot on K-ras oncogen detection. The results showed the sensitivity of CLMA was fund 83% and the specificity was 96%. On the other hand, the sensitivity of the membrane dot blot on K-ras oncogen detection increase to 95.5% and the specificity increase to 97.9%. In summary, this study has provided a foundation model on which we can develop a variety of diagnostic membrane array for related oncogenes and tumor suppress genes.