Ag490 Combined With Cisplatin On Proliferation And Apoptosis Of Human Cervical Carcinoma Hce1 Cell Function

Objects:To investigate whether AG490 combined with cisplatin reduced the growth of a human cervical carcinoma cell line HCEI cell through induction of apoptosis in vitro.Methods:(1)We cultured the human cervical carcinoma cell line HCEI cell.(2)Essay grouping and measure:HCEI 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,HCEI cell were cultured and treated by AG490 with 20umol/L,40umol/L and 80umol/L concentration for 24,48and72hours, by cisplatin with 20ug/mL,40ug/mL and 80ug/mL concentration for 24,48and72hours, by AG490 combined with cisplatin for 24,48and72 hours, the effect of AG490 on HCEI cells were analyzed by MTT assay, respectively. HCEI 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 HCEI 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 for24,48 and 72 hours, the apoptotic rate and cell cycle distribution were analyzed by flow cytometry after PI staining, respectively.Results:(1)HCEI cells became round, small, cell shrinkage, membrane blebbing and so on, after exposed to AG490.(2)After treated with AG490 with concentration about 20umcol/L,40umol/L,80umol/L 24 hours, HCEI cell inhibition rate discern is 10.6%±0.68%,18.1%±1.53%,34.3%±1.79%, after 48 hours, HCEI cell inhibition rate discern is29.1%±2.29%,45.3%±3.17%,67.0%±2.97%, after72hours, HCEI cell Inhibition rate discern is 40.5%±2.83%,66.3%±7.04%,82.5%±1.77%; After treated with cisplatin with concentration about 20ug/mL,40ug/mL and 80ug/mL 24 hours, HCEI cell inhibition rate discern is 4.6%±0.58%,7.5%±0.99%,15.1%±2.54%, after48hours, HCEI cell inhibition rate discern is 7.5%±0.48%,10.9%±1.37%,21.0%±3.07%, after 72 hours, HCEI cell inhibition rate discern is 15.1%±3.07%,43.1%±3.92%,60.4%±4.28%;After treated with 20umol/L AG49O combined with 20ug/mL cisplatin, HCEI cell inhibition rate discern is 31.2%±3.35%, MIT 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 inereased 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 20 umol/L AG490 48 hours, the G1-Phase cell fraction and the S-Phase cell fraction were 87.1%±3.12%, 9.9%±0.67%, after treated with 40ug/ML cisplatin 48 hours, the G1-Phase cell fraction and the S-Phase cell fraction were 3.1%±0.27%,93.2%±6.25%, after treated with 20 umol/L AG490 combine with 40ug/mL cisplatth AG490 combined with cisplatin.Conclusions (1)Janus Kinase inhibitor AG490 inhibited the growth the cervical careinoma cells by concentration and time dependent, The mechanism of which relates to its cell cyele arrest and apoptos in duction on cervical careinoma cells.(2)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.