白藜芦醇对人胃癌耐药细胞SGC-7901/5-FU生物学效应的影响及其作用机制

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（0 μmol/L、10 μmol/L、50 μmol/L、200 μmol/L、400 μmol/L）的RES分别处理SGC-7901/5-FU细胞24 h、48 h与72 h，其次用100 μmol/L浓度的RES与5-FU（5 μmol/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。

Effect of resveratrol on biological effects of human gastric cancer SGC-7901/5-FU cells and its mechanisms

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) and 5-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.