Genistein对人尿道鳞癌细胞增殖和凋亡的影响

Effect of genistein on cell proliferation and apoptosis in the human urethral squamous cancer cell line HUS-98

目的 观察Genistein(GEN)对人尿道鳞癌细胞系HUS 98细胞增殖和凋亡的影响。方法 MTT法检测细胞增殖 ,碘化丙啶 (PI)和AnnexinV FITC双染色的流式细胞分析方法检测凋亡 ,台盼蓝拒染法测定淋巴细胞活力 ,免疫细胞化学检测雌激素受体 (ER)蛋白的表达。结果 低浓度 (1× 10 11～ 1× 10 8mol/L)GEN促进HUS 98细胞的生长 ,处理 3d时 ,1× 10 8mol/L组的细胞数比对照组的增加 9% (P <0 .0 1) ;高浓度 (1× 10 7mol/L～ 5× 10 4mol/L)GEN则浓度依赖性地引起HUS 98细胞数目逐日减少 ,但是除 5× 10 4mol/L外的其余浓度对正常淋巴细胞的生长没有影响。 1× 10 6mol/LGEN作用于HUS 98细胞 2 4～ 4 8h ,凋亡细胞和坏死细胞的比率均增加 ,且有时间依赖性。HUS 98细胞表达ER蛋白。结论 高浓度GEN具有抑制ER阳性的人尿道鳞癌细胞的生长和诱导凋亡及坏死的作用 ,为今后用于尿道鳞癌的治疗提供了实验依据。

Objective To investigate the effect of genistein(GEN) on the proliferation, apoptosis and necrosis in a human urethral squamous cancer cell line HUS-98. Methods The effect of GEN on proliferation of HUS-98 cells was shown by MTT assay, the effect on cell viability was determined using trypan blue exclusion, cell apoptosis and necrosis were examined by PI and Annexin V-FITC double staining with flow cytometry, and immunocytochemistry was used to examine expression of estrogen receptor (ER) protein. Results GEN at low concentrations(1×10 -11-1×10 -8mol/L)enhanced growth of HUS-98 cells, on the 3rd day, cell number in 1×10 -8 mol/L GEN group was 9 % more than that of control(P<0.01);while GEN at high concentrations(1×10 -7mol/L-5×10 -4mol/L)could make the cell number decrease day after day in a concentration-dependent manner, but had no effect on the viability of normal lymphocytes except at 5×10 -4mol/L. After treatment with GEN at 1×10 -6mol/L for 24-48 h , both the percentage of the apoptosis cells and the necrosis cells increased in a time-dependent manner. Conclusion GEN at high concentrations can inhibit growth and induce apoptosis and necrosis in ER-positive human urotract cancer cells. The results provide evidence for the potential use of genistein in the treatment of human urethral squamous cancer.