【摘要】 目的探讨白藜芦醇(resveratrol,RES)对5-氟尿嘧啶(5-fluorouracil,5-FU)耐药胃癌细胞(SGC-7901/5-FU)生物学效应的影响及其作用机制。方法用不同浓度(0μmol/L、10μmol/L、50μmol/L、200μmol/L、400μmol/L)的RES分别处理SGC-7901/5-FU细胞24 h、48 h与72 h,其次用100μmol/L浓度的RES与5-FU(5μmol/L)单独或联合处理SGC-7901/5-FU细胞48 h。CCK-8法检测细胞增殖情况,流式细胞术检测细胞凋亡情况,免疫荧光染色法观察细胞形态。RT-PCR检测多药耐药基因MDR1以及凋亡相关基因Bcl2与Caspase 3的mRNA表达水平,Western blot检测P-gp、Bcl2和Caspase 3蛋白的表达水平。结果不同浓度RES均能抑制SGC-7901/5-FU细胞增殖,且具有时间与剂量依赖效应;RES组、5-FU组及联合组均能诱导SGC-7901/5-FU细胞凋亡,细胞出现凋亡样形态改变,其中联合组变化最明显(P<0.001);RES组和联合组中MDR1 mRNA与P-gp蛋白表达水平均下降(P<0.05),且联合组变化更明显;RES组、5-FU组及联合组中Bcl2 mRNA与蛋白表达水平均下降(P<0.001),Caspase 3 mRNA与蛋白表达水平均上升(P<0.001),其中联合组变化最明显。结论 RES通过下调多药耐药基因MDR1/P-gp蛋白表达抑制胃癌耐药细胞SGC-7901/5-FU细胞增殖并诱导细胞凋亡,降低细胞耐药性。更多还原

【Abstract】 Objective To investigate the effect of resveratrol(RES) on the biological effects of SGC-7901/5-FU cells and its mechanisms.Methods The SGC-7901/5-FU cells were treated with RES at concentrations of 0 μmol/L, 10 μmol/L, 50 μmol/L, 200 μmol/L and 400 μmol/L, respectively, for 24 h, 48 h and 72 h, and then SGC-7901/5-FU cells were treated, with RES(100 μmol/L) and5-FU(5 μmol/L), respectively or simultaneously, for 48 h. The cell proliferation was detected by CCK-8 method, the apoptosis by flow cytometry,and the cell morphology by immunofluorescence staining method. The mRNA expression levels of MDR1, Bcl2 and Caspase 3 were detected by RT-PCR, and the protein expression levels of P-gp, Bcl2 and caspase 3 were detected by Western blot.Results RES with different concentrations could inhibit the proliferation of SGC-7901/5-FU cells effectively, depending upon time and dose. The RES group, 5-FU group and combined group could induce apoptotic and morphology changes in SGC-7901/5-FU cells, which was most remarkable in the combined group(P<0.001). The expression levels of MDR1 mRNA and P-gp protein in RES group and combined group were decreased(P<0.05), which was more significant in combined group; the Bcl2 mRNA and protein expression levels were decreased in RES group,5-FU group and combined group(P<0.001), while Caspase 3 mRNA and protein expression levels were increased(P<0.001), which was more pronounced in the combined group.Conclusions RES can inhibit the proliferation of SGC-7901/5-FU cells by down-regulating the expression of drug-resistance related gene MDR1/P-gp protein and induce cell apoptosis to reduce drug resistance.更多还原