| Objective:To investigate the effects of apatinib on esophageal cancer cells in proliferation,migration and apoptosis,and investigate the possible mechanisms.Methods:MTT assay was used to measure the proliferation rate;transwell assay was used to determine the migration capacity;colony formation assay was used to assess the clone formation rate.The effect of apatinib on cell cycle and apoptosis was analyzed by flow cytometry.The expression levels of VEGF and VEGFR2 were measured by real-time quantitative PCR(qRT-PCR).The concentration of VEGF in the supernatant of esophageal cancer cell was assessed by enzyme-linked immunosorbent assay(ELISA).The expression levels of STAT3,p-STAT3,CHK2 and CDC2 were detected by western blot.Results : Apatinib inhibited the proliferation of esophageal cancer cell in a time-dependent(P<0.05)and concentration-dependent(P<0.05)manner.After 20,40μmol/L of apatinib intervention,the migration of KYSE-150 cells were 428.67±4.16,286.67±1.53,which was significantly lower than the control group(874.67±22.75),(P<0.05);the migration of ECA-109 cells were 1123.67±70.00,477.33±26.84,which was significantly lower than the control group(874.67±22.75),(P<0.05).After 20,40 μmol/L of apatinib intervention,the clone formation rates of KYSE-150 cell were(58.19±24.73)% and(29.10±22.40)%(P<0.05);the clone formation rates of ECA-109 cell were(61.12±17.21)% and(43.09±11.13)%(P< 0.05).After 20 μmol/L apatinib intervention,the apoptosis rates of KYSE-150,ECA-109 cells were(15.65±1.54)% and(49.26±1.62)%,which were significantly higher than that of control group [(3.49±0.74)%,(6.31±1.43)% ](P<0.05).In addition,apatinib increased the proportion of cells in G2 / M phase(P <0.05).After 20 μmol/L of apatinib intervention,the relative expressions of VEGF mRNA and VEGFR2 mRNA in KYSE-150 cell were(42.57±10.43)% and(36.09±10.82)%.The relative expressions of VEGF mRNA and VEGFR2 mRNA in ECA-109 cell were(75.68±9.37)% and(17.24±9.52)%,all of which were significantly decreased comparing to control group(P <0.05).After 20 and 40 μmol/L of apatinib intervention,concentration of VEGF inKYSE-150 cell were 766.48±114.27 pg/ml and 497.40±102.18 pg/ml,which were significantly decreased comparing to control group(967.41±57.75pg/ml)(P <0.05).Meanwhile,concentration of VEGF in ECA-109 cell were 675.21±46.69 pg/ml and598.95±47.60 pg/ml,which were significantly decreased comparing to control group(980.68±92.74pg/ml)(P <0.05).After 20 μmol/L of apatinib intervention,the gray value of A STAT3/A GAPDH 、A p-STAT3/A GAPDH 、A CHK2/A GAPDH、A CDC2/A GAPDH were0.62±0.09 、 0.36±0.13 、 0.47±0.12 and 0.48±0.08 respectively,which were significantly decreased comparing to control group(0.96±0.15 、 0.85±0.16 、0.91±0.20、0.78±0.15)(P <0.05).Conclusions : Apatinib can induce apoptosis of esophageal cancer cell KYSE-150 and ECA-109,and inhibit the cell proliferation,migration and colony formation.Moreover,apatinib can inhibit the tumor growth in esophageal carcinoma xenograft models.This inhibitory action of apatinib is related to the alterations in VEGF-related pathways such as JAK2/STAT3 pathway. |
