%0 Journal Article
%T Targeting of the AXL receptor tyrosine kinase by small molecule inhibitor leads to AXL cell surface accumulation by impairing the ubiquitin-dependent receptor degradation
%A Anja Weber
%A Markus Lauter
%A Robert Torka
%J Archive of "Cell Communication and Signaling : CCS".
%D 2019
%R 10.1186/s12964-019-0377-8
%X Increased AXL protein level after 24£¿h of low ¦ÌM treatment with selective AXL TKI. BMS777607 (BMS) treatment led to increased AXL protein levels in (a) Hs578T, (b) H292 and (c) MDA-MB231 cells. Western blots of AXL after treatment with a three-fold serial dilution of BMS from 20£¿¦ÌM to 1.01£¿nM are shown. ACTIN or GAPDH served as loading controls. A representative sample of multiple biological replicates is displayed. (d) Treatment with low ¦ÌM concentrations of BMS increased the viability of 3D cell spheroids. Cells were cultivated in a 3D matrix in the presence of BMS (0.001£¿¦ÌM to 12.5£¿¦ÌM). ATP measurements were performed after 72£¿h. All results were normalized to DMSO control values as indicated by a doted horizontal line. Low BMS concentrations between 0.097£¿¦ÌM and 0.78£¿¦ÌM increased significantly the ATP content of Caliper, MDA-MB231, H292 and H1792 cells in a rage of 110 to 125%, in contrast to Hs578T cells. BMS treatment had no impact on AXL mRNA transcription. 0.5£¿¦ÌM BMS treatment showed no change of AXL mRNA levels by RT-qPCR analysis within 4£¿h in (e) Hs578T, (f) H292 and (g) H1792 cells. Housekeeping genes GAPDH and ALAS served as normalization controls utilizing the ¦¤¦¤CT method. Two different primer sets were used namely, P3 and PB. Mean values and SEM of three independent experiments are show
%K RTK
%K AXL
%K TKI
%K Ubiquitin
%K Degradation
%K 3D spheroid
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6555758/