Dominant Negative Effect Of RON△170, A Splicing Product Of RON Receptor Tyrosine Kinase, On An Auto-phosphorylating Variant RON△160

ObjectiveTo investigate the roles of RON△170 receptor tyrosine kinase on an auto-phosphory lating variant RON△160 in 3T3 cell line.MethodsNIH-3T3 cell and 3T3-RONA160 cell were transfected with plasmid pcDNA3.1-RON△170 to construct 3T3-RON△170 and 3T3-RON△160RON△170 cell line; The tyrosine phosphorylation of RON was measured by Immunoprecipitation method; Cell growth and motile/invasive ability were tested by soft agar and migration assay.Results3T3-RON△170 and 3T3-RON△160RONA170 cell lines stably expressing RON△170 protein were constructed successfully; 3T3-RON△170 had no phosphorylated RON expression; phosphorylated RON expression of 3T3-RON△160RON△170 cell was lower than 3T3-RONA160 cell. Soft agar results showed growth cell clones of 3T3, 3T3-RON△160,3T3-RONA170 and 3T3-RON△160RON△170 were 12.33±2.08, 121.33±12.58,14.00±3.00 and 34.33±8.32,3T3-RONA160 compared to 3T3-RONA160RON△170,t=7.93, P< 0.05; Transwell migration assay results showed migrated cells of 3T3,3T3-RONA160,3T3-RONA170 and 3T3-RON△160RON△170 were 24.50±3.50,399.00±13.00,31.00±6.00 and 200.50±11.50,3T3-RON△160 compared to 3T3-RON△160RON△170, t=132.33, P< 0.05;ConclusionsRONA170 inhibits the tyrosine auto-phosphorylating, transfect and oncogenic ability of RONA160; RONA170 is a potential dominant negative variant of RON receptor tyrosine kinase.