GPRC5A-mediated inhibition of cell invasion and anchorage-independent growth of NSCLCs was repressed by tyrosine phosphorylation. A, The proliferation rate of H1792 cells stable transfected with vector, GPRC5A-WT, or -4 F were grown in 96-well plates in growth medium with 10% FBS with or without 100 ng/ml EGF for 24, 48, 72 , 96 and 120 hours. The number of cells in each well was measured by the MTT assay. B, The migration of H1792 stable transfectants as indicated was assayed. The photomicrographs of cells on the bottom side of the filter (migrated cells) (left) and the mean (and 95% confidence intervals) number of migrated cells was shown in the bar graph (right). The differences between the number of cells in EGF-treated GPRC5A-WT and -4 F stable transfectants in H1792 cells were not statistically significant (P >0.05; two-sided z test). C, anchorage-independent growth was assessed in GPRC5A-WT and -4 F transfected H1792 cells. The cells were resuspended in agarose/Matrigel and analyzed for colony formation over 2 weeks. The photomicrography of colonies (left), and the number of colonies (bar on right) were shown. The differences between the number of colonies in EGF-treated GPRC5A-WT and 4 F mutant H1792 cells were statistically significant (P <0.01; two-sided z test).