Exploring Of Time Effect Of SU11274 Impacting On Biological Behavior Of Esophageal Carcinoma Eca109 Cells

Objective:To study the impact of SU11274-c-Met protein inhibitor on the biological behavior of Eca109 cells, and via the potential relationships between its target-c-Met protein, c-met mRNA and clock mRNA to explore whether time effect exists during the treatment by SU11274 on Eca109 cells.Method:(1) Eca109 cells were cultured in vitro and c-Met protein was tested by immuncytochemistry. (2) Cell proliferation, movement, invasion, apoptosis rate and cell cycle were detected with MTT, cell scratch healing test, transwell assay and FCM after treated with SU11274 (20nM was used as the IC5o). (3) Clock and C-met mRNAs were assayed by RT-PCR. (4) The expression of c-Met protein at different time was detected by FCM. (5) The cell cycles after treated with SU11274 at different time points were detected by FCM.Result:(1) Eca109 cell expressed c-Met protein. (2) After the cells were treated with SU11274, the abilities of growth, migration and invasion were decreased contrast with those in control groups, and the apoptosis rates were higher than that in control groups, while the percentages of cells at S stage in DNA duplication were less. (3) RT-PCR results showed that expression of Clock mRNA was oscillating during 48h (P＜0.05). (4) The Clock and C-met mRNA expressed with circadian rhythms in 24h (P<0.01), and the expression in both of them showed similarity. (5) The c-Met protein expressed with oscillation in 24h (P<0.05) which was in accord with that of C-met mRNA. (6) The decreasing extent of proliferation index was different when Eca109 cells were treated by SU11274 at different time points.Conclusion:(1) The c-Met inhibitor SU11274 can inhibit Eca109 cells growing, infiltrating and migration, increase cell apoptosis rate and restrain cells transiting to S from Gl stage in cell proliferation cycle. (2) Eca109 cell cultured in vitro express Clock with rhythm. (3) Eca109 cells expressed C-met mRNA and c-Met protein with rhythms. (4) The inhibition rates of proliferation index were different when Eca109 cells were treated at different time points, suggesting that time effect exist in the treatment for esophageal cancer by SU11274 and be taken seriously.