A Novel siRNA-Based Approach to Study Mechanisms of Resistance/Action of a New Drug in Treatment of Breast Cancer. Addendum
Final rept. 30 Jul 2005-29 Jul 2006
UNIVERSITY OF MEDICINE AND DENTISTRY OF NEW JERSEY NEWARK
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The mechanism by which ET-743 YondelisTM, trabectedin exerts its antitumor activity is not understood. The goal of this study was to study the mechanism of actionresistance of ET-743 in breast cancer cells using a novel siRNA-based approach. Two primers representing the sense and anti-sense DNA template of the random siRNA inserts were cloned into the linearized pFIV-H1U6 siRNA expression vector. Out of 33 siRNA sequences obtained from vector transformed E.coli colonies, predicted that 67 of the siRNA template sequences would generate functional siRNA sequences and that 82 of the functional siRNA sequences were random. A pool of plasmids encoding the random siRNAs with a possible 105 to 106different siRNAs, either transduced with pseudo- lentiviral particles or transfected with electroporation, were tested in the MCF-7 breast cancer cell line. The cell line was then treated with a lethal dose of ET-743 and cytarabine, however no resistant colonies were obtained. Further scaling-up is required to fufill the goals of this project.
- Medicine and Medical Research