| Objects To investigate whether AG490 combined with cisplatin reduced the growth of a human cervical carcinoma cell line HCE1 cell through induction of apoptosis in vitro.To observe the effect of AG490 on the expression of P53.Methods(1)We cultured the human cervical carcinoma cell line HCE1 cell.(2)Essay grouping and measure:HCE1 cell were cultured and treated by AG490 with 20umol / L,40umol / L and 80umol / L concentrations for 24 hours respectively.Cell viability were measured by MTT assay.At the sametime,HCE1 cell were cultured and treated by AG490 with 20umol / L,40umol / L and 80umol / L concentration for 24,48 and 72 hours,by cisplatin with 20ug / mL,40ug / mL and 80ug / mL concentration for 24,48 and 72 hours,by AG490 combined with cisplatin for 24,48 and 72 hours,the effect of AG490 on HCE1 cells were analyzed by MTT assay,respectively.HCE1 cells were cultured and treated by AG490 with 20umol / L,40umol / L and 80umol / L concentrations for 24,48,72 hours,the apoptotic rate of HCE1 cell were analyzed by flow cytometry after PI staining,respectively.Treated with 20umol / L,40umol / L and 80umol / L AG490,20ug / mL,40ug / mL and 80ug / mL cisplatin,AG490 combined with cisplatin for 24,48 and 72 hours,the apoptotic rate and cell cycle distribution were analyzed by flow cytometry after PI staining,respectively.(3)P53 expression: HCE1 cells were treated with 20umol / L,40umol / L and 80umol / L AG490 for 48 hours,respectively.And the expression of P53 protein were detected by immunocytochemistry.During the experiments the control group's cells were treated by the same concentration DMSO,statistical analysis was used to check the significant differences among them.Results(1)HCE1cells became round,small,cell shrinkage,membrane blebbing and so on,after exposed to AG490.(2)After treated with AG490 with concentration about 20umol/L,40umol/L,80umol/L 24 hours,HCE1cell inhibition rate discern is 9.5%±0.66%,17.1%±1.51%,33.3%±1.77%,after 48 hours,HCE1cell inhibition rate discern is 28.0%±2.27%,45.2%±3.15%,66.0%±2.95%,after 72 hours,HCE1cell inhibition rate discern is 39.4%±2.81%,66.2%±7.02%,81.5%±1.78%; After treated with cisplatin with concentration about 20ug / mL,40ug / mL and 80ug / mL 24 hours,HCE1cell inhibition rate discern is 3.6%±0.56%,6.5%±0.98%,14.1%±2.52%,after 48 hours,HCE1cell inhibition rate discern is 6.5%±0.46%,9.9%±1.35%,20.0%±3.05%,after 72 hours,HCE1 cell inhibition rate discern is 14.1%±3.05%,42.1%±3.90%,59.4%±4.26%;After treated with 20umol/L AG490 combined with 20ug / mL cisplatin,HCE1 cell inhibition rate discern is 30.2%±3.34%,MTT assays showed that Janus Kinase inhibitor AG490 inhibited the growth of the cervical carcinoma cells by concentration and time dependent,and synergistic effect appeared when combined with cisplatin.(3)Flow cytometry results demonstrated that the cell cycle was redistributed:the G1-Phase cell fraction was conspicuous increased while the S-Phase cell fraction was significantly decreased after the cells were treated with AG490(P＜0.05).However,the result was opposite after treated with cisplatin.When treated with both of them,G1-Phase and S-Phase cell fraction were between the values of single agent.For example,after treated with 20umol/L AG490 48 hours,the G1-Phase cell fraction and the S-Phase cell fraction were 86.0%±3.01%,9.1%±0.66%, after treated with 40ug/mL cisplatin 48 hours,the G1-Phase cell fraction and the S-Phase cell fraction were 2.1%±0.26%,92.2%±6.24%,after treated with 20umol/L AG490 combine with 40ug/mL cisplatin 48 hours, the G1-Phase cell fraction and the S-Phase cell fraction were 13.2%±0.78%,80.5%±2.46%.(4)Both of AG490 and cisplatin could induce apoptosis in cervical carcinoma cells by a time and dose dependent way.The apoptotic index were significantly increased after treated with AG490 combined with cisplatin.(5)In contrast to control group,the P53 protein expression level obvious up-regulated in AG490 group(P＜0.05).Conclusions Janus Kinase inhibitor AG490 inhibited the growth of the cervical carcinoma cells by concentration and time dependent,the mechanism of which relates to its cell cycle arrest and apoptosis induction on cervical carcinoma cells.(2)Janus Kinase inhibitor AG490 up-regulates the expression P53 via inhibiting the JAK-STAT3 signaling pathway,further inhibits the growth of cervical carcinoma cells and promotes its apoptosis.(3)The apoptotic index was significantly increased after treated with AG490 combined with cisplatin,suggesting that the combination of them could exert synergistic antiproliferative effect on cervical carcinoma cells,the apoptotic effect of AG490 in cervival cancer cells were proposed to following by P53-overexpressing.
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