Tyrosine phosphorylations of both DAB1 and ABL are essential for invasion of colorectal cancer cells. Tyrosine phosphorylations of both DAB1 and ABL are.

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Tyrosine phosphorylations of both DAB1 and ABL are essential for invasion of colorectal cancer cells. Tyrosine phosphorylations of both DAB1 and ABL are essential for invasion of colorectal cancer cells. A, exogenous expression of DAB1 mRNA increased Matrigel invasion of RKO cells, and SRC/ABL dual inhibitor dasatinib (Das; at 0.01 μmol/L) showed a tighter suppressive effect on Matrigel invasion of RKO cells than the SRC inhibitor PP2 (at 10 μmol/L), either with or without additional expression of DAB1. Imatinib (Ima; 10 μmol/L) also showed significant inhibition of invasion (see B below). B, the ABL inhibitor imatinib blocks Matrigel invasion of RKO cells in a dose-dependent manner. C, ABL knockdown in human colorectal cancer cells inhibits Matrigel invasion. RKO cells were transfected with either of two independent siRNA sets (#1 or #2), each consisting of distinct siRNAs against ABL1 (siABL1-1 and -2) and ABL2 (siABL2-1 and -2). Ns, nonsilencing control. D, DAB1 increases ABL Tyr-kinase activity in human colorectal cancer cells. Lanes 1 to 3, HCT116 cells were transfected with the same amount of FLAG-tagged WT DAB1 (lane 2) or its 5YF mutant (lane 3), and their phospho-Tyr (pY) contents were determined by immunoprecipitation–Western blotting (IP-WB). Lanes 4 to 7, HCT116 cells were cotransfected with FLAG-tagged DAB1 (DAB1-FLAG) and HA-tagged WT ABL1b (ABL1b-HA). Note that the input amount of DAB1-FLAG cDNA for lanes 6 and 7 was the same as that for lanes 2 and 3. Lanes 8 to 11, HCT116 cells were cotransfected with the same set of DAB1 cDNA as in Lanes 4 to 7, and KD mutant of ABL1b. E, ABL is found in DAB1 immunoprecipitation (IP) of intestinal tumors in Apc/Aes mice as analyzed by Western blotting (left), whereas DAB1 is found in ABL IP, reciprocally (right). WB indicates antibodies used for detection of ABL or DAB1 by Western blotting. Data are presented as the mean of three independent experiments, with SD. *, P < 0.01. Masahiro Sonoshita et al. Cancer Discovery 2015;5:198-211 ©2015 by American Association for Cancer Research