尼美舒利和5-氟尿嘧啶联用对胃癌细胞的协同抑制作用及机制  

Synergistic inhibitory effect of nimesulide in combination with 5-fluorouracil on gastric cancer cells and its possible mechanisms

作　　者：张丽[1] 刘文忠[1] 陆红[1] 李恩灵[1] 萧树东[1] 

机构地区：[1]上海第二医科大学附属仁济医院上海市消化疾病研究所,200001

出　　处：《中华消化杂志》2005年第9期530-533,共4页Chinese Journal of Digestion

摘　　要：目的体外研究选择性环氧合酶-2(COX-2)抑制剂尼美舒利和5-氟尿嘧啶(5-FU)对胃癌细胞的抑制作用及机制。方法以胃癌细胞株MKN45、MKN28为研究对象,观察尼美舒利和5-FU单独或联合应用对细胞增殖、凋亡和细胞周期的影响。应用MTT法检测细胞增殖,流式细胞仪检测细胞凋亡(FITC-Annexin-V/PI双标记)和细胞周期,RT-PCR观察用药前后COX-2mRNA在两株细胞中的表达,Western免疫印迹法观察经两种药物单独和联合作用48h后细胞内凋亡相关蛋白Bax和Bcl-2的表达。结果在MKN45和MKN28细胞中均可观察到不同水平的COX-2mRNA表达,尼美舒利和5-FU联合应用可明显抑制COX-2mRNA表达。尼美舒利可抑制两株细胞的增殖并诱导凋亡。尼美舒利和5-FU具有协同抑制细胞增殖及诱导凋亡的作用,该作用与两种药物作用顺序无关,但在联用时作用最强。两药协同抑制增殖的作用主要通过协同杀伤和诱导凋亡而实现。5-FU增强了凋亡诱导蛋白Bax的表达,而尼美舒利则减少凋亡抑制蛋白Bcl-2的表达。两药联用可明显抑制胃癌细胞株生长。结论选择性环氧合酶-2抑制剂尼美舒利和5-FU通过抑制COX-2mRNA的表达及增强Bax/Bcl-2的表达比率诱导胃癌细胞凋亡,从而对胃癌细胞起到协同抑制增殖的作用。Objective To investigate the inhibitory effects of combined treatment of nimesulide, a selective cyclooxygenase-2, inhibitor, with 5-fluorouracil （5-FU） on gastric cancer cell lines and its possible mechanisms. Methods The human gastric cancer cell lines MKN45 and MKN28 were used in the study. After 48 hours treatment with nimesulide or 5-FU alone or in combination, the cells growth was determined by MTT assay. Flow cytometry and FITC-Annexin-V/PI kit were used to determine the effect of drugs on tbe cell cycle and the apoptosis of gastric cancer cells. The expression of COX-2 mRNA was detected by RT-PCR. The expressions of apoptosis-associated protein Bax and Bcl-2 were detected by Western blotting. Results Differcnt level of COX-2 mRNA expression was observed in MKN45 and MKN28 cell lines. Treatment of the two cell lines whh nimesulide in combination with 5-FU can significantly reduce the expression of COX-2 mRNA. Nimesulide could inhibit the growth and induce apoptosis of gastric cells. Comhined treatment of nimesulide with 5-FU resulted in a synergistic effect of inhibiting growth and inducing apoptosis. The synergistic inhibiting effect on cell growth was irrespective of treatment sequence, but the highest cytotoxity was obtained when the cell lines were trcated with two drugs simultaneously. Treatment with 5-FU enhanced expression of the pro-apoptotic gene Bax, while nimesulide reduced expression of the anti-apoptotic gene Bcl-2, resulting in a significantly higher ratio ot Bax-to- Bcl-2. Conclusions The combined treatment of nimesulide with 5-FU results in synergistic inhibiting effect on growth of gastric cancer cell lines, and inducing apoptosis by enhanced Bax-to-Bcl-2 ratio and suppression of COX-2 mRNA expression may be the mechanisms.

关 键 词：环氧合酶-2抑制剂 胃癌细胞 凋亡 5-氟尿嘧啶 

分 类 号：R735.2[医药卫生—肿瘤]