[Changes of phenotypes associated with activated Src by overexpressed PTPalpha in NIH3T3 cells].
NIH3T3 cells transfected with inducible expression vectors harboring the protein tyrosine phosphatase alpha (PTPalpha) gene were used as a model to study the mechanism of tumor formation, by which the changes of phenotypes in inducible cells were studied. When NIH3T3 cells transfected with PTPalpha were induced for 24 hours, the expression levels of PTPalpha in inducible cells was higher than those in uninducible cells as judged by RT-PCR and Western blotting; the expression level of Src in inducible cells was the same as those in uninducible cells, but activity of Src kinase in inducible cells was higher than those in uninducible cells.The level of phosphated tyrosine in Src was reduced in inducible cells. The malignant changes of phenotypes in the inducible cells were verified by flow cytometry and electron microscope. The results show that 24 h induction of PTPalpha could initiate the transformation of NIH3T3 cells transfected with PTPalpha inducible expression vectors, and the transformation may be associated with activated Src where pTyr(527) in the C tail was dephosphorylated by increased expression level of PTPalpha in the cells.
Yang
Jin-Song
JS
Department of Biochemistry, Lab of Molecular Biology and Research Center for Human Gene Therapy, Shanghai Second Medical University, Shanghai 200025, China. sschen@shsmu.edu.cn
Zheng
Xin-Min
XM
Chen
Shi-Shu
SS
D016475:3T3 Cells; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015153:Blotting, Western; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D002466:Cell Nucleolus / Q000648:ultrastructure; D002467:Cell Nucleus / Q000648:ultrastructure; D002471:Cell Transformation, Neoplastic; D003593:Cytoplasm / Q000648:ultrastructure; D004318:Doxycycline / Q000494:pharmacology; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D051379:Mice; D008854:Microscopy, Electron; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D019061:src-Family Kinases; D017027:Protein Tyrosine Phosphatases; D004318:Doxycycline