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目的研究塞来昔布在体外抑制人结肠癌细胞Caco-2生长增殖及其抗肿瘤的相关分子机制。方法体外培养人结肠癌细胞Caco-2,分组为正常组(无任何干预)及塞来昔布组。四甲基偶氮唑盐(MTT)法检测塞来昔布在相同浓度下,不同时间对于胃癌细胞增殖的影响,并计算半数抑制浓度(IC50)值;反转录聚合酶链式反应(RT-PCR)法检环氧化酶-2(COX-2)、基质金属蛋白酶9(MMP-9)的mRNA的表达影响。结果 MTT结果显示:相同干预浓度塞来昔布抑制人胃癌细胞增殖,其24 h,48 h,72 h的IC50分别为:99.519±10.355μmol/L、71.546±6.446μmol/L、59.622±15.999μmol/L;RT-PCR结果显示:人结肠癌细胞Caco-2正常对照组及塞来昔布干预组COX-2mRNA灰度值分别为:0.808±0.021,0.101±0.002(t=19.037,P=0.000);MMP-9 mRNA灰度值分别为:0.798±0.031,0.190±0.002(t=18.987,P=0.002)。结论塞来昔布可能通过抑制COX-2、MMP-9的mRNA的表达,从而抑制结肠癌细胞增殖。
Objective To study the molecular mechanisms of celecoxib inhibiting Caco-2 proliferation and anti-tumor activity in human colon cancer cells in vitro. Methods Human colon cancer Caco-2 cells were cultured in vitro and divided into normal group (without any intervention) and celecoxib group. MTT assay was used to detect the effect of celecoxib on the proliferation of gastric cancer cells at the same concentration and at different times, and the half-value inhibition concentration (IC50) value was calculated. Reverse transcriptase-polymerase chain reaction -PCR method was used to detect the mRNA expression of cyclooxygenase-2 (COX-2) and matrix metalloproteinase 9 (MMP-9). Results MTT results showed that the IC50 of celecoxib at the same concentration for 24 h, 48 h and 72 h were 99.519 ± 10.355μmol / L, 71.546 ± 6.446μmol / L, 59.622 ± 15.999μmol / L. The results of RT-PCR showed that the gray value of COX-2 mRNA in Caco-2 normal control group and celecoxib group were 0.808 ± 0.021,0.101 ± 0.002 (t = 19.037, P = 0.000 ); The gray value of MMP-9 mRNA were 0.798 ± 0.031,0.190 ± 0.002 (t = 18.987, P = 0.002). Conclusion Celecoxib inhibits the proliferation of colon cancer cells by inhibiting the mRNA expression of COX-2 and MMP-9.