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目的观察三羟基异黄酮(Genistein)联合放疗对人肺腺癌细胞A549增殖凋亡及其Ape1/Ref1蛋白的影响。方法 1组单用0、10、30、60、90μmol/L的Genistein处理A549细胞48 h,另1组用同样浓度的Genistein预处理24 h后予以直线加速器放疗8 Gy,继续培养24 h后用噻唑蓝比色法(MTT)检测各组活细胞数,确定最适Genistein联合放疗的浓度。用流式细胞仪检测对照组、单纯放疗组(IR 8 Gy)、单纯Genistein组(GEN 30μmol/L)、Genistein+IR组(GEN30μmol/L+IR 8 Gy)的细胞凋亡;采用Western blot法检测各组细胞中Ape1/Ref1蛋白表达。结果 30、60、90μmol/L的Genistein联合放疗比单用相应浓度的Genistein处理细胞增殖抑制率高(P<0.05),但低浓度10μmol/L联合放疗与单纯放疗比较差异不明显[(26.67±1.26)%vs(24.70±0.72)%,P=0.102]。IR组、Genistein+IR组早期凋亡率较对照组明显增高[(30.88±7.63)%、(42.98±1.46)%vs(6.80±0.98)%,P<0.05],加药组较对照组无明显统计学差异[(12.96±4.99)%vs(6.80±0.98)%,P=0.143)];与对照组、单纯放疗组、单纯加药组比,Genistein+IR组中Ape1/Ref1的表达明显下调。结论三羟基异黄酮可增加放疗对A549细胞的增殖抑制作用,可能与减少Ape1/Ref1蛋白的表达有关。
Objective To observe the effects of Genistein combined with radiotherapy on proliferation and apoptosis of human lung adenocarcinoma A549 cells and its effect on Ape1 / Ref1 protein. Method 1 The A549 cells were treated with 0, 10, 30, 60 and 90μmol / L of Genistein for 48 h. The other group was pretreated with the same concentration of Genistein for 24 h and then treated with linear accelerator 8 Gy. After 24 h, The number of viable cells in each group was determined by MTT assay to determine the concentration of Genistein in combination with radiotherapy. Flow cytometry was used to detect apoptosis of control group, radiotherapy group (8 Gy), Genistein group (GEN 30 μmol / L) and Genistein + IR group (GEN 30μmol / L + IR 8 Gy) Ape1 / Ref1 protein expression in each group was detected. Results Genistein combined with radiotherapy at 30, 60 and 90μmol / L had a higher rate of cell proliferation inhibition than Genistein alone (P <0.05), but no significant difference was found between low dose of 10μmol / L and radiotherapy alone [(26.67 ± 1.26)% vs (24.70 ± 0.72)%, P = 0.102]. The early apoptotic rate in IR group and Genistein + IR group was significantly higher than that in control group [(30.88 ± 7.63)%, (42.98 ± 1.46)% vs (6.80 ± 0.98)%, P <0.05] (12.96 ± 4.99)% vs (6.80 ± 0.98)%, P = 0.143). The expression of Ape1 / Ref1 in Genistein + IR group was significantly higher than that in control group, radiotherapy alone group and simple drug group Down. Conclusion Trihydroxy isoflavone can increase the radiosensitivity of A549 cells, which may be related to the decrease of Ape1 / Ref1 protein expression.