A Pleiotrophy-Specific SiRNA And Its Effect On PTEN～(-/-) MEF241 Cells

Objective: Pleiotrophin is a kind of secreted growth factor and signals with diverse functions, including cell growth, migration and angiogenesis. The study aims to design a siRNA which could inhibit malignant phenotype of Pten^-/- MEF241 cell lines and inhibit the tumorigenesis and its development.Methods: A group of double strand oligonucleotide fragments were synthesized and cloned into pSilencerTM 3.1-H1 hygro vector. Lipofectamine 2000 transfection and hygromycin B screening were used to establish Ptn stable silenced Pten^-/- MEF241 cell lines. Northern blot was used to measure the expression of Ptn and the cell growth rate was measured to analyze the effect of Ptn on Pten^-/- MEF241 cells'proliferation; Western blot was used to analyze the signal pathway; tumorigenesis model in nude mice was also established to observe the change of malignant phenotypes of the Pten^-/-M?EF241 cells after gene silencing of Ptn.Results: Three Ptn-specific siRNAs have been designed and constructed into pSilencer^TM 3.1-H1 hygro vector, and one of them was proved to be able to effectively depress Ptn gene in Pten^-/- MEF241cells.