The animals were checked frequently for tumor progression

Figure 1D showed the quantity of t Src, p Src, t EGFR and p EGFR analyzed by software of ImageJ. The cell via bility of group A, B and C did not show any significant dif ference by various concentration of dasatinib in sk Hep1 and Huh 7 cells. Although we showed serum affected the cell Amuvatinib 臨床試験 proliferation, it couldnt affect the response of HCC cells to dasatinib. The effects of dasatinib on Src and downstream targets were detected by western blotting in dasatinib treated cells. The expression ratio of individual phosphor protein to B actin was quantified by ImageJ software. We analyzed the protein inhibition level in HCC cells when treated with dasatinib at the dosage of 1uM. In general, there was a significant correlation between the IC50 of dasatinib and the inhib ition of p Src, p Akt and p FAK576 577 by dasatinib.

In all 3 sensitive cell lines, sk hep1, Li 7 and PLC PRF 6, the sensitivity to dasatinib was significantly correlated with p Src and P FAK576 577 in hibition by dasatinib. 5 out of 9 HCC cell lines including all sensitive cell lines had a significant correlation between p Src inhibition and p FAK576 577 inhibition by dasatinib. P Src AT-406 代理店 inhibition and p Akt inhibition by dasatinib were also showed significant correlation in 5 HCC cell lines. We didnt find any significant inhibition of Stat3 and MAPK42 44 activities in all cell lines by dasatinib at the dosage of 1uM and below. Individually, sk Hep1, the most sensitive to dasatinib growth inhibition, showed only moderate inhibition of p Src, p FAK576 577 and p Akt by dasatinib at the dos age of 1uM.

Even though dasatinib completely inhibited the expression of p Src at 0. 1uM in Li 7 cells, it only moderately reduced the p FAK576 577 activity without inhibiting p Akt. both sk Hep1 and Li 7 expressed lower p Src and p Src t Src. It suggested that dasatinib may affect other signal pathway and inhibiting other protein kinase or growth factors to regulate cell growth in these two cell AG-490 臨床試験 lines. PLC PRF 6 was the only dasatinib sensitive cell line that co overexpressed t Src and t EGFR, higher baseline expression of p Src and lower p Src t Src. In order to investigate whether dasatinib would affect EGFR signaling pathway, the activity of EGFR was tested too. The p Src, p FAK576 577, p FAK861 and p Akt were significantly inhibited by dasatinib at 0. 1uM, p EGFR1068 was inhibited at 10uM.

No inhibition of t Src expression by dasatinib at all. It appeared at lower concentration of dasatinib there was a slight increase of p Src. The mechanism of such difference is unknown. However, the ratio of p Src t Src of control vs dasatinib treatment did not have any significant difference. Huh 7 was the least sensitive to dasatinib and very little level of p Src was detected before dasatinib treatment but inhibition of p Src can be demonstrated by dasatinib. In this cell line, dasatinib not only could not reduce p FAK at both 576 577 and 861 sites, but also increased the level of them suggesting Src dependant signaling pathway is not crucial in the regulation of oncogenic pro cesses for Huh 7 cells. HT 17 is one of the most resistant cell lines to dasatinib, but is sensitive to gefitinib.