Synthetic lethal and shRNA screen in oral cancer cell lines
Date Issued
2008
Date
2008
Author(s)
Lin, Meng-Yi
Abstract
We developed a straightforward surfection method for cells wherein DNA/polyethylenimine/gelatin complexes are coated on the surface of 96 well plates and stored in -80℃ until cells are ready for transfection. Comparing to most of the conventional transfection, surfection can be performed in the presence of serum. Importantly, DNA/polyethylenimine complexes promote cell adhesion to the plates. Also, long-term storage of the plates did not reduce the transfection efficiency nor it had any effects on the cell toxicity. Because of the stability of complexes, surfection enables large-scale tansfection thus providing a reproducible, cost-effective and versatile tool for parallel screening of proteins, gene regulatory elements and virus production used in diverse applications.n Taiwan, the incidence and the mortality of oral squamous cell carcinoma (OSCC) were ranked fourth in men among all malignancies. It is well documented that cigarette smoking and areca nut chewing contribute to the risk of oral squamous cell carcinoma (OSCC). The role of phosphorylated Akt (p-Akt) in oral carcinogenesis induced by nicotin and alkaline environments was investigated. eguelin, a rotenoid of the flavonoid family, is able to decrease tumor incidence in animal models for lung, colon, mammary and skin carcinogenesis through Akt inhibition. We used surfection to produce lentivirus shRNA to infect oral cancer cell SAS. High-throughput RNAi screening along Akt pathway was carried out in oral cancer cell to analyze cell apoptosis and growth. High throughput screening with Degulin and RNAi is of particular value as it’s not only for analyzing a compound’s mechanism of action and understanding of unwanted side effects but also for identifying potential gene targets for developing sensitizing agents for existing drugs.
Subjects
oral cancer cell
shRNA
deguelin
SDGs
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