As_2O_3对肾癌786-0细胞5-氟尿嘧啶药物敏感性的研究

RESEARCH ON THE DRUG SENSITIVITY OF 786–0 RENAL CARCINOMA CELLS TO ARSENIC TRIOXIDE

目的探讨As2O3与5-氟尿嘧啶(5-FU)联合应用对肾癌细胞786-0细胞株细胞增殖、细胞凋亡及X连锁凋亡抑制蛋白(XIAP)表达的影响。方法取对数生长期786-0细胞,MTT比色法检测As2O3和/或不同浓度5-FU对细胞生长抑制作用;流式细胞仪检测细胞凋亡率;RT-PCR法及蛋白质印迹法检测XIAP mRNA表达及XIAP蛋白质表达的变化。结果不同浓度的5-FU分别作用786-0细胞后,细胞生长受到抑制,但无明显的时间、浓度相关性;2μmol/L As2O3与不同浓度5-FU联合应用对786-0细胞的增殖抑制率随作用时间、药物浓度的增加逐渐升高,与单药相比较(P<0.01),两药相互作用系数显示为协同作用。48h检测2μmol/L As2O3与不同浓度5-FU联合用药处理的786-0细胞的凋亡率明显增加,与单药相比(P<0.01);XIAP mRNA表达明显减少,与单药相比(P<0.01);2μmol/L As2O3与400μg/ml 5-FU联合应用48h XIAP蛋白表达亦明显减少,与单药相比(P<0.01)。结论 As2O3联合5-FU可以明显提高肾透明细胞癌的化疗敏感性,抑制786-0细胞增殖,诱导肿瘤细胞凋亡,这一作用与抑制786-0细胞XIAP基因的表达有关。

Objective To explore the effect of the combined use of arsenic trioxide and 5-fluorouracil on the pro- liferation, apoptosis and the expression of XIAP of 786 - 0 renal carcinoma cells. Methods 786 - 0 cells at log phase were chosen, and the growth inhibiting effect of As203 and/or different levels of 5-fluorouraeil was observed with chromatometry. Flow cytometry was used to detect the rate of apoptosis. RT-PCR and immunoblotting method were used to detect the expressions of XIAP mRNA and the changes in the expressions of XIAP. Results The differ- ent concentrations of 5-fluomuracil had growth inhibiting effect on cell growth but showed no obvious dependability in time and concentration. The ratio of cytostasis of 786-0 ceils caused by the the combined use of 2？ mol of arse-nic trioxide and 5-fluorouracil of different concentrations increased with time and the concentration of drugs, and there existed significant difference between the combined use of drugs and the drugs used separately（ P 〈0.01 ）. The coefficient of interaction belonged to synergistic action. The detection performed after 48 hours showed the rate of apop- tosis of 786-0 cells caused by the combined use of drugs in-creased obviously, and there existed significant difference between the combined use of drugs and the drugs used separately（ P 〈0.01 ）. The expressions of XIAP mRNA decreased obviously, and there existed significant differ- ence between the combined use of drugs and the drugs used separately（ P 〈0.01 ）. The expressions of XIAP after 48 hours decreased obviously, and there existed significant difference between the combined use of drugs and the drugs used separately（ P 〈 0.01 ）. Conclusion The chemotherapy sensitivity of renal carcinoma can be enhanced by the the combined use of arsenic trioxide and 5-fluorouracil to inhibit the growth of 786-0 cells and to induce the apoptosis of tumor ceils, which is related to the expressions of XIAP genes of 786-0 calls.