艾瑞昔布联合奥沙利铂对裸鼠人结肠癌LOVO细胞移植瘤模型的细胞凋亡作用机制研究

Effect and mechanism of imrecoxib combined with oxaliplatin on apoptosis of human colon cancer LOVO cells transplanted in nude mice model

目的探讨艾瑞昔布联合奥沙利铂对裸鼠人结肠癌LOVO细胞移植瘤模型的细胞凋亡的影响及与生存素(Survivin)、半胱氨酸天冬氨酸蛋白酶-3(Caspase 3)的关系。方法用裸鼠构建人结肠癌LOVO细胞裸鼠皮下移植瘤模型,将造模成功裸鼠随机分为4组:空白对照组、艾瑞昔布组、奥沙利铂组和联合用药组,每组16只。空白对照组(0.9%NaCl);艾瑞昔布组(艾瑞昔布100 mg·kg-1·d-1,灌胃,1次/天);奥沙利铂组(奥沙利铂30 mg·kg-1·d-1,静脉注射,1次/周);联合用药组(艾瑞昔布组+奥沙利铂组),连续给药21 d。计算瘤体体积,缺口末端标记法法检测瘤体凋亡指数(AI)%,酶联免疫吸附法检测裸鼠血清Survivin和Caspase-3浓度,逆转录-聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)检测瘤体Survivin和Caspase-3基因表达,免疫组化检测瘤体Survivin和Caspase-3蛋白表达。结果空白对照组、艾瑞昔布组、奥沙利铂组和联合用药组的瘤体体积分别为(1210±230),(953±205),(897±174)和(651±155)mm3;这4组的AI%分别为(10.35±1.52)%,(32.60±2.26)%,(40.57%±2.74)%和(55.23±3.40)%。这4组的血清Survivin浓度分别为(16.15±1.33),(13.76±1.26),(13.25±1.20)和(10.24±1.17)μg·L-1;这4组的Caspase-3浓度分别为(5.62±1.16),(13.06±1.23),(14.44±1.26)和(22.16±1.38)μg·L-1;基因和蛋白结果的趋势与血清浓度一致。上述指标:3个给药组与空白对照组比较,差异均有统计学意义(均P<0.05);艾瑞昔布组、奥沙利铂组与联合用药组比较,差异均有统计学意义(均P<0.05),说明联合用药组疗效最好。结论艾瑞昔布联合奥沙利铂可能通过调控Survivin、Caspase-3表达,抑制结肠癌移植瘤细胞凋亡,增强奥沙利铂的抗肿瘤效果。

Objective To investigate the effect of imrecoxib combined with oxaliplatin on apoptosis of human colon cancer LOVO cells transplanted in nude mice and its relationship with Survivin and Caspase-3. Methods Nude mice were selected to construct human colon cancer LOVO cells subcutaneously transplanted tumor model. According to the random number method,the nude mice successfully modeled were randomly divided into 4 groups:blank control group,imrecoxib group,oxaliplatin group,joint group,16 mice in each group.Blank control group(0.9% NaCl,gavage,1 time/day),imrecoxib group(imrecoxib 100 mg·kg-1d-1,gavage,1 time/day),oxaliplatin group(oxaliplatin 30 mg·kg-1d-1,intravenous injection,1 time/week),joint group(imrecoxib group + oxaliplatin group),continuous administration for 21 d. Tumor volume was calculated. Serum concentrations of Survivin and Caspase-3 were detected by enzyme linked immunosorbent assay. Survivin and Caspase-3 mRNA expression were detected by reverse transcription polymerase chain reaction(RT-PCR). Tumor apoptosis index(AI,%) were detected by TdT-mediated dUTP nick-end labeling. Results Tumor volume in blank control group,imrecoxib group,oxaliplatin group,and joint group were(1210 ± 230),(953 ± 205),(897 ± 174) and(651 ± 155) mm3;the AI% in the 4 groups were(10. 35 ± 1. 52) %,(32. 60 ± 2. 26) %,(40. 57% ± 2. 74) % and(55. 23 ± 3. 40) %;Serum Survivin concentrations in the 4 groups were(16. 15 ± 1. 33),(13. 76 ± 1. 26),(13. 25 ± 1. 20) and(10. 24 ± 1. 17) μg · L-1;Serum Caspase-3 concentrations in the 4 groups were(5. 62 ± 1. 16),(13. 06 ± 1. 23),(14. 44 ± 1. 26) and(22. 16 ± 1. 38)μg·L-1. The trend of gene and protein results is consistent with that of serum concentration. Compared with the blank control group,the difference of the factors in the three treatment groups were significant(all P < 0. 05);compared between joint group and irexeb group,oxaliplatin group,the difference of the factors were significant(all P < 0. 05).The results showed that the joint group had the best effect. Conclusion Imrecoxib combined with oxaliplatin may inhibit the apoptosis of colon cancer transplanted tumor cells by regulating the expression of Survivin and Caspase-3,and enhance the anti-tumor effect of oxaliplatin.