Project title
微小RNA 於轉化生長因子BETA第一型受體調控滋養層細胞侵襲與子癲前症相關之研究
Internal ID
NSC101-2314-B002-082
Principal Investigator
Start Date
August 1, 2012
End Date
July 12, 2013
Investigators
Partner Organizations
Project Web Site
The Role of Mirna in the Regulation of Typei Tgfbeta Receptor-Mediated Trophoblast Invasion and the Pathogenesis of Preeclampsia = 微小RNA 於轉化生長因子BETA第一型受體調控滋養層細胞侵襲與子癲前症相關之研究
Description
The invasion of trophoblast is a key step for the development of embryo and placenta. Shallow invasion of trophoblast may cause preeclampsia, while deep invasion results in placenta accreta of pregnancy. In our last project, we have demonstrated that the trophoblast invasion is regulated by the TRI expression. However, how TRI expression exerts their signals to regulate the invasion capacity of trophoblast is largely unknown. In this project, we’ll apply the emerging technique of micro RNA (miRNA) to understand how miRNA regulate the TRI-mediated trophoblast invasion. Using miRNA array, we hope we can identify the new regulators of trophoblast invasion, and then using them as biomarkers to early detect preeclampsia. In the first year, we use 3 different trophoblastic cell lines and the primary trophoblast from the first-trimester curettage. We will clone 6 sets of RNAi targeting to TRI into the lentiviral plasmid, then deliver into the target cell. These RNAi may result in increasing invasion and migration of the trophoblast. After confirming by real-time PCR and Western blotting that TRI is really decrease, we extract total RNA from the cells then sent for miRNA analysis. The most significant upregulated and downregulated miRNA (before and after treatment) will be picked up by bioinformatics. In the project of the second year, we overexpress the target miRNAs by plasmid transfection, and also knockdown them using miRNA sponge in the other. After confirmation by phenotypic and functional assays by invasion chamber and time-lapse video microscopy, we can verify these miRNAs do regulate the TRI-mediated trophoblast invasion. In the last year, we’ll extract total RNA from the blood of the patients from preeclamptic pregnancies and healthy control before the day of delivery. The samples are sent to real-time PCR to compare the expressions of our target miRNAs identified in the first 2 years. After statistics, the most significant changes of miRNA will be picked up. They may serve the biomarkers of preeclampsia and probably lead to the development of the personalized treatment.