艾瑞昔布联合5-氟尿嘧啶对裸鼠结肠癌移植瘤侵袭和转移的影响

目的探讨艾瑞昔布联合5-氟尿嘧啶(5-Fu)对结肠癌HT-29细胞裸鼠移植瘤侵袭和转移的影响以及与环氧化酶-2(COX-2)、血管内皮生长因子-C(VEGF-C)、基质金属蛋白酶-9(MMP-9)的关系。方法 40只裸鼠构建人结肠癌HT-29细胞裸鼠皮下移植瘤模型,随机分为4组,每组10只,对照组(0.9%NaCl);艾瑞昔布组100 mg/(kg·d)];5-Fu组(20 mg/kg);联合用药组艾瑞昔布100 mg/(kg·d),5-FU 20 mg/kg],艾瑞昔布ig给药,1次/d,5-Fu ip给药,每3天1次,连续给药14 d。实验结束时测量各组裸鼠瘤体大小,并计算抑瘤率;酶联免疫吸附试验(ELISA)检测COX-2、VEGF-C、MMP-9血清浓度,实时荧光定量PCR(qRT-PCR)检测COX-2、VEGF-C、MMP-9的m RNA表达,免疫组化法检测瘤体微血管密度(MVD),Westernblotting法检测COX-2、VEGF-C、MMP-9蛋白表达。结果艾瑞昔布组、5-Fu组、联合用药组与对照组比较,抑瘤效果显著,其中以联合用药组抑瘤效果最佳(P<0.05)。艾瑞昔布组、5-Fu组、联合用药组较对照组裸鼠血清COX-2、VEGF-C、MMP-9浓度、瘤体COX-2、VEGF-C、MMP-9的m RNA和其蛋白表达、瘤体MVD均显著降低(P<0.05),其中以联合用药组降低最显著(P<0.05)。结论艾瑞昔布联合5-Fu可协同抑制结肠癌裸鼠移植瘤侵袭和转移,增强5-Fu的抗肿瘤效果,其作用机制可能与下调COX-2、VEGF-C、MMP-9表达有关。

Effects of ericoxib combined with fluorouracil on invasion and metastasis of colon cancer xenografts in nude mice

Objective To investigate the effect of ericoxib combined with fluorouracil on the invasion and metastasis of colon cancer HT-29 cells in nude mice, and with cyclooxygenase-2 (COX-2), vascular endothelial growth factor-C (VEGF-C), matrix metalloprotein-9 (MMP-9) relationship. Methods Forty nude mice were used to construct a subcutaneous xenograft model of human colon cancer HT-29 cells in nude mice. Model nude mice were randomly divided into 4 groups, 10 mice in each group. Four groups were control group (0.9% Nacl), ericoxib group ericoxib 100 mg/(kg.d)], fluorouracil group (5-Fu 20 mg/kg), and combination group ericoxib 100 mg/(kg.d), 5-Fu 20 mg/kg]. Ericoxib was administered by intragastrically, once daily, 5-Fu was administered by intraperitoneal injection, once 3 days, continuous administration for 14 days. Tumor size were measured and of tumor inhibition rate were calculated at the end of the experiment. Enzyme linked immunosorbent assay (ELISA) was used to detect concentrations of COX-2, VEGF-C and MMP-9 in serum of nude mice. mRNA expression of COX-2, VEGF-C and MMP-9 were detected by Real-time quantitative PCR (Real-time PCR). MVD of tumor was detected by immunohistochemical method. COX-2, VEGF-C and MMP-9 protein expression were analyzed by Western blotting. Results Compared with the control group, the tumor inhibition effect of the ericoxib group, the fluorouracil group and the combination group were remarkable, and the anti-tumor effect was the best in the combination group (P<0.05). Compared with the control group, COX-2, VEGF-C, MMP-9 concentrations in serum, mRNA and protein expression of tumor COX-2, VEGF-C, MMP-9, tumor MVD of nude in the ericoxib group, fluorouracil group, and combination group were reduced. Among them, anti-tumor effect of the combination group was the most significant (P<0.05). Conclusion Erexib combined with fluorouracil can synergistically inhibit the invasion and metastasis of human colon cancer xenografts in nude mice, and enhance the anti-tumor effect of fluorouracil. The mechanism may be related to the down-regulation of COX-2, VEGF-C and MMP-9 expression.