Effect of tyrosine kinase inhibitor BGJ398 on proliferation,apoptosis and migration of human hepatocellular cancer Huh-7 cells and its mechanism / 中国生物制品学杂志

@#Objective To evaluate the effect of tyrosine kinase inhibitor BGJ398 on the proliferation,apoptosis and migration of human hepatocellular cancer Huh-7 cells and explore the mechanism.Methods The effects of 10 tyrosine kinase inhibitors on the survival of Huh-7 cells were detected by MTT assay,and the sensitivity of Huh-7 cells to BGJ398 was analyzed by single-target kinetic equation and biphasic kinetic equation respectively.Huh-7 cells were added with 10,30 and 90 nmol/L BGJ398 respectively,and the control group(without drugs)was set.The effects of BGJ398 on the apoptosis and cell cycle of Huh-7 cells were detected by flow cytometry after culturing at 37℃for 24 h,the effect on the migration ability was detected by wound healing assay and the effect on the expression of multiple pathway-related proteins was detected by Western blot.Results All of 10 tyrosine kinase inhibitors inhibited the proliferation of Huh-7 cells,among which Huh-7 cells were most sensitive to BGJ398 and the IC_(50)was(0.020±0.013)μmol/L;The response of Huh-7 cells to BGJ398 was composed of two phases with F_1 accounted for 92.8%(K_(d1)was 36 nmol/L)and F_2 accounted for 7.2%(K_(d2)>1 000μmol/L).Compared with the control group,the apoptosis rate and the percentage of Huh-7 cells in G1 phase increased significantly in 30 and 90 nmol/L BGJ398 groups(t=-6.407～-4.459,each P<0.05),while the percentage of Huh-7 cells in S phase decreased significantly in 10,30 and 90 nmol/L BGJ398 groups(t=2.982,7.859 and 12.425,respectively,each P<0.05);After 24 and 48 h of scratching,the scratch area of 30 and 90 nmol/L BGJ398groups decreased significantly(t=5.376～18.197,each P<0.05);The expression levels of phosphorylated fibroblast growth factor receptor(FGFR)and phosphorylated extracellular signal-regulated kinase 1/2(Erk1/2)protein decreased significantly in 30 and 90 nmol/L BGJ398 groups(t=4.015～6.729,each P<0.01).Conclusion BGJ398 can inhibit the proliferation and migration of human hepatocellular cancer Huh-7 cells,induce apoptosis and cell cycle arrest,which might be achieved by inhibiting FGFR phosphorylation and MAPK signaling pathway.BGJ398 is expected to be a potential agent for the treatment of hepatocellular cancer.

Inhibitory Eefects of the novel tyrosine kinase inhibitor BGJ398 against human leukemic cell line KG-1 cells / 中华血液学杂志

Objective: To explore the effects and possible mechanisms of the novel pan-FGFR inhibitor BGJ398 on KG-1 cells in vitro. Methods: Effects of BGJ398 on cells proliferation were detected by CCK-8, the apoptosis was assessed by Annexin V-FITC. Reverse transcriptionquantitative polymerase chain reaction (q-PCR) analysis was used to detect the expression of apoptosis-related genes B cell lymphoma-2 (Bcl-2) and caspase-3. Western blotting analysis was performed to explore the proteins expression levels of Bcl-2, caspase-3 and the expression of p-AKT, p-S6K, p-ERK and FGFR1. Results: BGJ398 effectively inhibited cell proliferation by dose-dependent manners. BGJ398(1.4 µmol/L) induced apoptosis of KG-1 cells by 36.4%, compared with 4.5% in the control group(P<0.001). Treatment with BGJ398 at 1.4 µmol/L led to significant increases in the expression levels of caspase-3, and decreases in the expression of Bcl-2 (P<0.005). In accordance with these results, Western blot analysis further confirmed the increased expression of Bcl-2 protein along with elevated caspase-3 activity. In addition, BGJ398 markedly down-regulated FGFR1OP2-FGFR1 fusion protein, p-AKT and p-S6K expression, but not p-ERK expression. Conclusion: Novel pan-FGFR inhibitor BGJ398 substantially suppressed KG-1 cell growth and induced apoptosis by inhibiting the expression of FGFR1, p-AKT, p-S6K and regulating apoptosis-related proteins.

Inhibitory Eefects of the novel tyrosine kinase inhibitor BGJ398 against human leukemic cell line KG-1 cells / 中华血液学杂志

Objective@#To explore the effects and possible mechanisms of the novel pan-FGFR inhibitor BGJ398 on KG-1 cells in vitro.@*Methods@#Effects of BGJ398 on cells proliferation were detected by CCK-8, the apoptosis was assessed by Annexin V-FITC. Reverse transcriptionquantitative polymerase chain reaction (q-PCR) analysis was used to detect the expression of apoptosis-related genes B cell lymphoma-2 (Bcl-2) and caspase-3. Western blotting analysis was performed to explore the proteins expression levels of Bcl-2, caspase-3 and the expression of p-AKT, p-S6K, p-ERK and FGFR1.@*Results@#BGJ398 effectively inhibited cell proliferation by dose-dependent manners. BGJ398(1.4 µmol/L) induced apoptosis of KG-1 cells by 36.4%, compared with 4.5% in the control group(P<0.001). Treatment with BGJ398 at 1.4 µmol/L led to significant increases in the expression levels of caspase-3, and decreases in the expression of Bcl-2 (P<0.005). In accordance with these results, Western blot analysis further confirmed the increased expression of Bcl-2 protein along with elevated caspase-3 activity. In addition, BGJ398 markedly down-regulated FGFR1OP2-FGFR1 fusion protein, p-AKT and p-S6K expression, but not p-ERK expression.@*Conclusion@#Novel pan-FGFR inhibitor BGJ398 substantially suppressed KG-1 cell growth and induced apoptosis by inhibiting the expression of FGFR1, p-AKT, p-S6K and regulating apoptosis-related proteins.