Proteomics Approach to Identify microRNA-372 Target in Human Lung Adenocarcinoma Cell

• Main Authors: Yu-Chieh Huang, 黃毓傑
• Other Authors: 周綠蘋
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/59602505872116775195

碩士 === 國立臺灣大學 === 生物化學暨分子生物學研究所 === 96 === MicroRNAs are 18–24 nt single-strand, small non-coding RNAs that can act as endogenous RNA interference. They can negatively regulate the expression of hundreds of their target genes by translational repression or mRNA cleavage through partial complementary to the 3’-UTR of these targets. MicroRNAs are involved in a wide range of biological functions, such as cellular proliferation, differentiation and apoptosis. Recent evidence indicates that microRNAs may function as tumor suppressors or oncogenes, thus alteration in microRNA expression may play a critical role in tumorigenesis and cancer progression. Until now, the molecular basis of miRNA-mediated gene regulation and the effect of these genes on tumor growth remain largely unknown because of our limited understanding of miRNA targets. Lung cancer, predominant non-small-cell lung cancer (NSCLC), is the leading cause of cancer death in Taiwan for recent years because of the symptoms of delayed-diagnosis, malignancy, metastatic capacity and highly-mortality rate. The evidence obtained by Yu. et. al demonstrate that five-microRNA signature can predict survival in lung cancer patients. One of them is miRNA-372 which can significantly increase the invasion ability of cancer cells, and may play significant roles in the metastasis process. In the previous studies, miRNA-372 was reported to act as oncogene in testicular germ cell tumors by direct inhibition of the tumor suppressor LATS2 expression, thus permitting proliferation and tumorigenesis. The major interest of this study is to find out miRNA-372 targets and investigate the cellular mechniams involved in miRNA-372 overexpression. We used the 2D-DIGE analyze the differential protein expression between the lung adenocarcinoma cell line CL1-0 that stably over-express miRNA-372 and vector only. Eighteen proteins were down-regulated and twelve proteins were up-regulated after miRNA-372 overexpression. Using target prediction programs, we found seven proteins may have miRNA-372 target site. The differential expressed proteins were analysed by the signal transduction database tool and found them involved in MAPK pathway and cytoskeleton remodeling. The enhanced invasion ability may be caused by these pathway. Among the protein in these networks, we found more twenty proteins that have potential miRNA-372 target site. In the future, these miRNA-372 regulations and their physiological roles need to be further validated.