T. Of the four inhibitors reached, the h HIGHEST 324,674 bcr-abl pathway inhibitory effect on cell proliferation in both HT29 and SW480 cells. In addition, we observed that treatment of EGFR / HER2/HER4 had a negligible inhibitory effect on both cell lines, with 47 4% of the cells at 20 lm survived in the HT29 cells, and 52 3% of the cells survived to 20 lm in SW480 cells. By generating dose-response curves for each cell line by MTT assay, we calculated the IC 50 values of the four ITC. Both HT29 and SW480 were strongly inhibited by the irreversible inhibitor EGFR 324,674, with an IC50 of 1.96 ml for the HT29 cells and LM 1.04 SW480 cells. However, both cell lines were less sensitive to AG1478 and GW583340 with 35.90 IC50, s in the range of 9.11 to LM. For EGFR/HER2/HER4-Hemmer-Therapie, a increased Hte IC50 LM 63.
71 was observed for HT29 and SW480 cells at LM 88.34. Overall, these results indicate that, was compared with the other three inhibitors, the irreversible EGFR inhibitor 324674 the gr Te inhibitory effect on HT29 and SW480 cell proliferation. 3.2. Irreversible EGFR inhibitors strongly HT29 and SW480 324 674-cell apoptosis, the observation that strongly inhibited 326 474 HT29 and SW480 cell proliferation suggested that TKI can effectively induce apoptosis. To this end, HT29 and SW480 cells were treated with or without 1 or 2 meters ITK 324 674, AG1478, or GW583340 EGFR/HER2/HER4 inhibitors. Forty-eight hours after treatment, cell apoptosis by annexin V binding assay was assessed followed by flow cytometry.
In both HT29 and SW480 cells, the treatment of 326,474 treatment at a final concentration of LM 2 significantly induces cell apoptosis, w During a treatment with the three other ITC at the same concentration had a lower influence on cell apoptosis. Under treatment with 2 ml of 326 474 in the HT29 cell apoptosis rate reached 18.74% of the total number of cells, w While the rate is below 10% in the other three treatments ITC. In SW480 cells, 2 ml of 326 474 treatment was entered Born a rate of 19.38% apoptotic cells, w While the other three ITC treatment led to a rate of apoptotic cells by 12%. These results indicate that irreversible EGFR inhibitors Fnd 324674 efficient cell apoptosis as inhibitors AG1478 or GW583340 EGFR/ErbB2/ErbB4 Promoted. 3.3.
Irreversible EGFR inhibitor blocked EGFR phosphorylation and downstream 324,674 events is known that upon activation, the activated EGFR dimerization of the kinase activity of t and tyrosine residue 1068 autophosphorylated rapidly. This autophosphorylation then the activation of the downstream Rtigen signaling proteins, comprising MAPK, Akt and ERK, which leads to a result of DNA synthesis and cell proliferation. Given the observation that the irreversible EGFR inhibitor treatment 324 674 cell proliferation and apoptosis also greatlyinhibited, we investigated whether it could EGFR autophosphorylation and suppress its downstream events. In both HT29 and SW480 cells, EGF treatment significantly solo h Here pY EGFR 1068th Therefore, signals downstream proteins, including Akt and ERK increased pp Ht. But additionally to introduce Tzlicher irrev