Effects Of 5-Fu Combined With Nimesulide On The Expression Of VEGF In Human Esophageal Carcinoma EC109 Cell Line

ObjectiveThrough investigating the effects of NIM(a cyclooxygenase 2 inhibitor),5-FU, and their combination on the expression of VEGF in human esophageal carcinoma EC109 cell line,we hope to primary explore 5-FU,NIM inhibit angiogenesis mechanism of esophageal carcinoma.Methods1.Nimesulide(100,200,400μmol/ml) and 5-FU(100,200,400μmol/ml)were used.EC109 cell lines were divided into control group(RPMI1640 which contains only 0.2% DMSO),Nimesulide treated group,5-FU treated group, and Nimesulide combined with 5-FU group.The inhibition on the proliferation of EC109 cells respectively treated with Nimesulide ,5-FU, and their combination were detected by MTT .The best effection group be choosed to next steps.2.EC109 cells were treated with control group, 400μmol/ml of NIM, 400μmol/ml 5-FU and their combination for 24h, and their apoptosis rates were detected by FCM. 3.EC109 cells were treated with control group, 400μmol/ml of NIM, 400μmol/ml 5-FU and their combination for 24h, then extracting the RNA.Reverse transcriptionpolymerase chain reaction(RT-PCR) was used to detect transcription of VEGF mRNA.4.EC109 cells were treated with control group, 400μmol/ml of NIM, 400μmol/ml 5-FU and their combination for 24h, then extracting the VEGF protein. The expression level of VEGF protein was detected by western blot.Results1.Different concentration of 5-FU and NIM could inhibit the growth of EC109 cells.Comparing to the control group, the survival rate of EC109 cells was significant descended along with increasing the drug dose(P<0.05).However,the depression effect of the inhibition rate of Nimesulide combined with 5-FU were significantly higher than the single NIM or 5-FU single treated (P <0.05).So,we choose 400μmol/ml of NIM and 400μmol/ml 5-FU continue next steps.2.Apoptosis rate of NIM combined with 5-FU were significantly higher than the single Nimesulide or 5-FU single treated (P<0.05).3.Comparing to the control group,both NIM and 5-FU could decrease the mRNA expression of VEGF.But,when the two drugs combined , the mRNA expression of VEGF were obviously decreased (P<0.05). 4. NIM combined with 5-FU significantly reduced VEGF protein expression,as compared with the single Nimesulide and 5-FU treated (P<0.05).Conclusions1.NIM and 5-FU could inhibit the growth of esophageal carcinoma cells EC109 when they single used,and the depression effect is obviously depend on the drug dosage. The inhibition rate and apoptosis rate of Nimesulide combined with 5-FU were significantly higher than the single Nimesulide or 5-FU single treated.2.Both NIM and 5-FU could decrease the protein and mRNA transription activity of VEGF,and compared with the control group,the depression effect became more and more stronger by the two drugs combined.3.The combined effect of NIM with 5-FU significantly reduced VEGFmRNA and protein ,as compared with the single effect Nimesulide and 5-FU. The mechanism of depression effect may be depend on regulating the exprssion of VEGF.