The Effect Of AG490 Combined With Gemcitabine On The Growth Of Human Pancreatic Cancer Cells In Vitro

Objective: To investigate the effects and mechanism of AG490 , Gemcitabine, their combination and different drug schedule on the proliferation and apoptosis, the expressions of the STAT3, p-STAT3, CyclinD1, Bcl-xl, Bax, and Survivin was detected under different treatments in human pancreatic cancer cell lines in vitro.Methods: The experiment was divided into six groups according to different drug treatment: the control group, the AG490 group, the Gemcitabine group, the AG490+Gemcitabine group, the AG490 followed by Gemcitabine group and the Gemcitabine followed by AG490 group. The cell proliferation was evaluated by MTT assay. The flow cytometry was used to observe the cell apoptosis. The transcriptional and translational expressions of STAT3, CyclinD1, Bcl-xl, Bax and Survivin were examined by Western blot and RT-PCR.Results: The cell growth rates of the AG490 group and the Gemcitabine group were (2.20±0.25) and (2.30±0.22) respectively, that were significantly lower than that of the control group(3.78±0.42). The apoptosis proportion of the AG490 group and the Gemcitabine group were (35.40±3.08)% and (34.64±1.38)% respectively, that were significantly higher than that of the control group (16.49±1.45)%. The expression of p-STAT3 protein of the AG490 group was significantly less than that of the control group (P<0.05). The expression of mRNAs and proteins of CyclinD1, Bcl-xl, and Survivin were less than that of the control group significantly as well. But the expression of mRNA and protein of Bax were significantly higher than that of the control group. The results of the Gemcitabine group were similar to that of the control group. The cell growth rate of the AG490 combined with Gemcitabine group was significantly lower than that of the AG490 or the Gemcitabine group (P<0.05). The apoptosis proportion of the AG490 combined with Gemcitabine group was significantly higher than that of the AG490 or the Gemcitabine group (P<0.05).The cell growth rate of the Gemcitabine followed by AG490 group was significantly lower than that of the AG490 followed by Gemcitabine group (P<0.05).The apoptosis proportion of the Gemcitabine followed by AG490 group was significantly higher than that of the AG490 followed by Gemcitabine group (P<0.05).Conclusion: Gemcitabine, AG490 or their combined treatment all have a significantly inhibitory effect on human pancreatic cancer cells proliferation and a significantly inducible apoptosis. AG490 could reduce the expressions of CyclinD1, Bcl-xL and Survivn, increase the expression of Bax, by blocking the activation of JAK and STAT3, inducing inhibition of the proliferation and promoting the apoptosis in pancreatic cancer cells. AG490 combined with Gemcitabine showed a synergic effect. The effect of the Gemcitabine followed by AG490 group is superior to that of the AG490 followed by Gemcitabine group.