山慈菇对甲状腺癌SW579细胞增殖及凋亡的影响

Effect of cremastra appendiculata makino on proliferation and apoptosis of thyroid cancer SW579 cells

目的探讨山慈菇对甲状腺癌SW579细胞增殖及凋亡的影响。方法采用不同浓度(10、20、40、80、160 mg/ml)的山慈菇提取液作用于甲状腺癌SW579细胞48 h,同时以只加培养液的处理组为对照组,采用噻唑蓝(MTT)法检测SW579细胞的增殖抑制率,计算半数抑制浓度(IC50)。收集经10 mg/ml和20 mg/ml山慈菇处理48 h的SW579细胞和只加培养液处理48 h的对照组SW579细胞,采用流式细胞仪检测SW579细胞的凋亡率,采用蛋白质印迹(Western blot)法检测SW579细胞中B细胞淋巴瘤因子-2(Bcl-2)的表达水平。结果 MTT检测结果显示,山慈菇浓度为0、10、20、40、80、160 mg/ml时,对SW579细胞的增殖抑制作用呈先上升后下降的趋势,山慈菇浓度为20 mg/ml时的增殖抑制率最高,经计算IC50为11.59 mg/ml。流式细胞仪检测结果显示,经10 mg/ml和20 mg/ml山慈菇处理的SW579细胞的凋亡率分别为(25.56±1.62)%和(58.85±2.26)%,均高于对照组的(8.05±1.05)%,差异均有统计学意义(P﹤0.05)。Western blot检测结果显示,经10 mg/ml和20 mg/ml山慈菇处理的SW579细胞中Bcl-2蛋白表达水平均低于对照组。结论山慈菇能够抑制甲状腺癌SW579细胞的增殖并诱导其凋亡,其作用机制可能与下调Bcl-2蛋白表达有关。

Objective To investigate the effects of cremastra appendiculata makino on proliferation and apoptosis of thyroid cancer SW579 cells. Method Thyroid cancer SW579 cells were treated with different concentrations（10, 20, 40,80, 160 mg/ml） of cremastra appendiculata makino extract for 48 h, and the control group was treated with only culture medium, the inhibition rate of SW579 cell proliferation was detected by MTT method, and the half inhibitory concentration（IC50） was calculated. The apoptosis rates of SW579 cells which were treated for 48 h by cremastra appendiculata makino with 10 mg/ml and 20 mg/ml concentrations, and the apoptosis rate of SW579 cells in control group, were all detected by flow cytometry. The expression level of Bcl-2 protein in SW579 cells was detected by Western blot. Result MTT test results showed that the proliferation inhibition rate of SW579 cells was first increased and then decreased along with cremastra appendiculata makino concentration in 0, 10, 20, 40, 80, 160 mg/ml, the inhibition rate reached the maximum at the 20 mg/ml cremastra appendiculata makino concentration, and the IC50 was 11.59 mg/ml. Flow cytometry results showed that the apoptosis rates of SW579 cells treated by cremastra appendiculata makino with 10 mg/ml and 20 mg/ml concentrations were（25.56＋1.62）% and（58.85＋2.26）%, respectively, which were significantly higher than that of the control group（8.05 ＋ 1.05）%（P〈0.05）. Western blot result showed that the expression levels of Bcl-2 protein in SW579 cells treated by cremastra appendiculata makino with 10 mg/ml and 20 mg/ml concentrations were significantly lower than that of the control group. Conclusion Cremastra appendiculata makinocan inhibit the proliferation and induce the apoptosis of thyroid cancer SW579 cells, and its mechanism may be related to the down-regulation of Bcl-2 protein.