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目的研究塞来昔布在体外对人结肠癌细胞COLO205生长增殖及对环氧化酶2(COX-2)、金属蛋白酶9(MMP-9)mRNA表达的影响。方法体外培养人结肠癌细胞COLO205,分组为正常对照组和塞来昔布干预组,MTT法检测正常对照组和塞来昔布组抑制率,塞来昔布在不同时间且相同浓度的条件下,不同时间对于结肠癌细胞增殖的影响,测得抑制率并算得半抑制浓度(IC50)值;RT-PCR检测COX-2、MMP-9的mRNA的表达影响。结果 MTT结果显示:相同浓度的塞来昔布干预结肠癌COLO205细胞,分别在24 h,48 h,72 h测算IC50结果为:96.620±9.044μmol/L、71.561±5.706μmol/L、58.982±11.069μmol/L,塞来昔布浓度提高和干预时间延长,结肠癌COLO205细胞活力下降。RT-PCR结果显示:正常对照组与塞来昔布干预组COX-2 mRNA灰度值分别为:0.832±0.012,0.113±0.001,干预组塞来昔布干预细胞的COX-2mRNA表达降低,所应显示的条带消失,无表达率,两组比较差异有显著性(t=18.051,P=0.000);正常对照组与塞来昔布干预组MMP-9 mRNA灰度值分别为:0.756±0.050,0.171±0.001,干预组MMP-9 mRNA表达明显降低,条带消失,无mRNA表达,两组比较差异有显著性(t=17.286,P=0.001)。结论体外实验表明:塞来昔布抑制结肠癌COLO205细胞增殖,通过抑制COX-2及MMP-9的mRNA的表达来实现抗肿瘤作用。
Objective To investigate the effect of celecoxib on the growth and proliferation of human colon carcinoma cell line COLO205 and the expression of cyclooxygenase 2 (COX-2) and metalloproteinase 9 (MMP-9) mRNA in vitro. Methods COLO205 human colorectal cancer cells were cultured in vitro and divided into normal control group and celecoxib intervention group. MTT assay was used to detect the inhibition rate of normal control group and celecoxib group. Celecoxib at different time and concentration , The effect of different time on the proliferation of colon cancer cells, inhibition rate was measured and calculated half-inhibitory concentration (IC50) value; RT-PCR detection of COX-2, MMP-9 mRNA expression. Results MTT results showed that IC50 of colon cancer COLO205 treated with celecoxib at the same concentration were 96.620 ± 9.044μmol / L, 71.561 ± 5.706μmol / L and 58.982 ± 11.069, respectively at 24 h, 48 h and 72 h μmol / L, increasing the concentration of celecoxib and prolonging the time of intervention, the activity of COLO205 cells in colon cancer decreased. RT-PCR results showed that the gray value of COX-2 mRNA in the normal control group and celecoxib intervention group were 0.832 ± 0.012 and 0.1313 ± 0.001 respectively, and the expression of COX-2 mRNA in the intervention group decreased significantly The expression of MMP-9 mRNA in the normal control group and celecoxib group was significantly lower than that in the control group (t = 18.051, P = 0.000) 0.050,0.171 ± 0.001. The expression of MMP-9 mRNA in the intervention group was significantly lower than that in the control group (P <0.001). There was no expression of MMP-9 mRNA in the intervention group. There was significant difference between the two groups (t = 17.286, P = 0.001). Conclusion In vitro experiments show that celecoxib inhibits the proliferation of human colon cancer COLO205 cells and achieves anti-tumor effect by inhibiting the expression of COX-2 and MMP-9 mRNA.