塞来昔布对胃癌MGC-803细胞RECK、MMP-2、MMP-9基因表达的影响

Celecoxib down-regulates RECK expression and up-regulates MMP-2 and MMP-9 expression in human gastric cancer cell line MGC-803

目的:研究非甾体类抗炎药(NSAID)塞来昔布对胃癌细胞MGC-803的抑制作用和对RECK、MMP-2、MMP-9基因表达的影响,以探讨塞来昔布的抗肿瘤机制.方法:培养胃癌MGC-803细胞,实验用不同浓度的塞来昔布(25、50、100μg/L)分别处理MGC-803细胞不同时间(12、24 h、48 h),无血清培养基饥饿24 h达同步化后,MTT(噻唑蓝比色法)观察MGC-803细胞增殖;RT-PCR法检测细胞周期调控因子MMP-9、MMP-2、RECK mRNA的表达;Western blot法检测MMP-9、MMP-2、RECK mRNA蛋白的表达.结果:M T T法显示塞来昔布能抑制胃癌MGC-803细胞增殖,作用12 h时,随着塞来昔布处理浓度的增高,其抑制作用不明显,组间比较差异不显著(P>0.05),在作用24、48 h时呈浓度依赖性(25-100μg/L),在25、50、100μg/L作用浓度呈时间依赖性(24-48 h).塞来昔布在100μg/L浓度作用细胞48 h时抑制.RT-PCR法检测结果显示,在12 h的作用时间点,随着塞来昔布处理浓度的增高,RECK基因及MMP-2、MMP-9 mRNA变化不是很明显,组间比较无显著差异(P>0.05),在12 h后,塞来昔布能增加胃癌细胞RECK mRNA和蛋白表达,作用呈浓度(25-100μg/L)和时间依赖性(24-48h),MMP-2、MMP-9表达则下降.Western blot法检测结果显示,作用12 h时,随着塞来昔布处理浓度的增高,其RECK蛋白及MMP-2,MMP-9蛋白改变不明显,组间比较差异不显著(P>0.05),在12 h以后,塞来昔布能增加RECKmRNA和蛋白表达并减少MMP-2、MMP-9表达,且作用呈浓度依赖性和时间依赖.结论:塞来昔布可抑制人胃癌MGC-803细胞增殖与转移;其可能是通过上调RECK基因,进而下调MMP-2、MMP-9来抑制胃癌细胞MGC-803的增殖与转移.

AIM： To explore the antitumor mechanisms of celecoxib by investigating the impact of non-ste- roidal anti-inflammatory drug （NSAID） celecoxib on the expression of RECK, MMP-2 and MMP-9 in human gastric cancer cell line MGC-803.METHODS： MGC-803 cells were starved in serum-free medium for 24 h and treated with different concentrations of celecoxib （25, 50, and 100 μg/L） for different durations （12, 24,and 48 h）. Cell proliferation was determined by MTT assay. The expression of MMP-9, MMP-2 and RECK mRNAs and proteins was detected by RT-PCR and Western blot, respectively.RESULTS： Celecoxib inhibited MGC-803 cell proliferation in a concentration-dependent man- ner at 24 and 48 h but not at 12 h. The inhibitory effect was also time-dependent. Cell prolifera- tion was mostly inhibited after treatment with celecoxib at a concentration of 100 μg/L for 48 h. The expression of RECK, MMP-2 and MMP-9 mRNAs showed no significant differences （all P 〉 0.05） among cells treated with different con- centrations of celecoxib for 12 h. After 12 h, celecoxib increased RECK mRNA expression in a concentrationand time-dependent manner, but decreased the expression of MMP-2 and MMP-9. Similar results were also obtained for RECK, MMP-2 and MMP-9 protein expression.CONCLUSION： Celecoxib inhibits the prolifera- tion and metastasis of MGC-803 cells possibly via mechanisms associated with up-regulation of RECK expression and down-regulation of MMP-2 and MMP-9 expression.