环氧化酶-2选择性抑制剂NS-398逆转HCT-8/FU细胞多药耐药及其机制探讨

Reversion effect of COX-2 selective inhibitor NS-398 on multi-drug resistance of HCT-8/FU cells and its mechanism

目的观察环氧化酶-2(COX-2)选择性抑制剂NS-398对结肠癌细胞HCT-8/FU多药耐药(MDR)的逆转作用,并初步探讨其作用机制。方法通过CCK-8法检测NS-398对HCT-8/FU的耐药逆转倍数,应用免疫细胞化学法检测NS-398作用前后细胞膜P-gp表达情况,流式细胞术检测细胞早期凋亡。结果 NS-398作用HCT-8/FU细胞24 h后,细胞对5-FU的敏感性显著增加,浓度为10μmol/L和20μmol/L时耐药逆转倍数为3.55和10.73倍,呈现剂量依赖性,20μmol/L作用24 h后免疫细胞化学法检测P-gp表达减少,流式细胞术检测到NS-398 10μmol/L、20μmol/L作用24 h,细胞早期凋亡率分别为(11.95±0.325167)%及(21.49±0.690217)%,均验证了该细胞对5-FU重新恢复敏感性。结论环氧化酶-2选择性抑制剂NS-398可逆转HCT-8/FU细胞多药耐药,一定浓度范围内呈现剂量依赖关系,无毒剂量的NS-398亦可具有逆转耐药的作用,其机制可能通过抑制P-gp的表达及促进细胞凋亡起作用。

【Objective】 Observe the multi-drug resistance（MDR） reversal of cyclooxygenase-2（COX-2） selective inhibitor NS-398 on colon cancer cells HCT-8/FU,and explore its mechanism preliminarily.【Methods】 detect the reversal of multi-drug resistance on HCT-8/FU by NS-398 through the method of CCK-8,detect the expression of P-gp before and after the effect of NS-398 through the immunocytochemical method,detect the apoptosis by flow cytometry（FCM）.【Results】 24 hours later after NS-398＇s effect on HCT-8/FU cells,the cells＇ sensitiveness to 5-FU increases remarkably;with the concentration of 10μmol/L and 20μmol/L,the reversal of multi-drug resistance are 3.55 and 10.73 respectively,and the dose dependent is presented;the expression of P-gp decreases after being effected by 20μmol/L for 24 hours through detection by immunocytochemical method;with the effect of NS-398 10μmol/L and 20μmol/L for 24 hours,apoptosis rate are 11.95±0.325167 % and 21.49±0.690217% respectively,which is detected by flow cytometry（FCM）.They all verify the inhibition of 5-FU to cells.【Conclusion】 Cyclooxygenase-2 selective inhibitor NS-398 can reverse multi-drug resistance of HCT-8/FU cell,and within certain range of concentration,the dose dependent relationship is presented.Non-toxic NS-398 also has the function of reversal resistance and it plays its role by inhibiting the expression of P-gp and promoting the apoptosis.