环氧化酶-2活性抑制对人膀胱癌T24细胞Survivin、Smad2、VEGFmRNA及其蛋白表达的影响

Effects of the cyclooxygenase-2 activity inhibition on the expression of Survivin, Smad2, VEGF mRNA and protein in human bladder cancer cell line T24 in vitro

目的观察环氧化酶-2(COX-2)活性抑制后,人膀胱癌T24细胞中Survivin、Smad2、VEGFmRNA及其蛋白表达的变化,探讨COX-2及其特异性抑制剂尼美舒利(NIM)对人膀胱癌生物学行为影响的可能机制。方法体外培养的人膀胱癌T24细胞受终浓度为90μmol/L的NIM作用后,采用ELISA法检测细胞分泌PGE2变化,半定量RT-PCR法、WesternBlot法检测COX-2、Survivin、Smad2、VEGFmRNA及其蛋白表达变化。结果COX-2活性抑制后T24细胞分泌PGE2较对照组明显减少(P<0.05);实验组Survivin、VEGFmRNA及其蛋白表达较对照组减弱(P<0.05),Smad2mRNA及其蛋白表达增强(P<0.05),而COX-2mRNA及其蛋白表达无显著变化(P>0.05)。结论COX-2及其特异性抑制剂NIM可能通过PGE2调控凋亡负性调控基因Survivin、信号传导相关的Smad2基因及血管形成相关的VEGF基因的表达,进而影响膀胱癌的生物学行为。

Objective To observe the change of Survivin, Smad2, VEGFmRNA and their protein in T24 cell lines by inhibiting activated COX-2 to investigate potential mechanism of COX-2 and the effect of selective COX-2 inhibitor nimesulide on the biological behavior of human bladder cancer. Methods The T24 cell line was cultured in vitro, and exposed to nimesulide （90μmol/L）. PGEz level was detected by ELISA. Expression of COX-2, Survivin, Smad2, VEGFmRNA and their protein was examined by semiquantitative reverse transcriptase-polyerase chain reaction（RT-PCR） and Western blot analysis. Results PGE2 production, and the expression of survivin and VEGF decreased more in the experimental group than in the control group（P〈0.05）. Expression of Smad2 increased in the experimental group（P〉0.05）, but expression of COX-2 in T24 did not change. Oonclusion COX-2 and the selective COX-2 inhibitor nimesulide may affect the biological behavoir of bladder cancer through regulating the expression of Survivin, Smad2, and VEGF by PGE2.