葫芦素B对人结肠癌细胞株LS174-T的生长抑制作用

Inhibitory effect of cucurbitacin B on proliferation of human colon cancer cells

目的 检测葫芦素B对于人结肠癌细胞的抑制作用.方法 0.01、0.10、1.00、10.00、100.00 μmol/L浓度的葫芦素B分别作用于LS174-T细胞24、48、72 h.细胞计数试剂盒（CCK-8）法检测细胞增殖,流式细胞仪检测细胞周期.荧光定量逆转录聚合酶链反应（FQ-PCR）检测转录信号转导子与激活子3 （STAT3）、半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3、及Ten mRNA水平.结果 CCK-8结果显示,0.01 μmol/L和100.00 μmol/L的葫芦素B作用24 h,细胞增殖抑制率分别为（5.22±2.02）％、（73.90±1.85）％.100.00 μmol/L的葫芦素B作用24和72 h,细胞增殖抑制率分别为（73.90±1.85）％、（98.84±0.51）％（P＜0.05）.流式细胞仪检测发现,与对照组比较,1.00 μmol/L浓度处理组处于G2/M期细胞的比例升高[（6.33±0.74）％比（30.43±4.09）％],Go/G1期细胞比例下降[（78.50±4.72）％比（43.20±5.65）％,P＜ 0.05].FQ-PCR结果显示,0.10μmol/L和10.00 μmol/L浓度组作用24 h,STAT3 mRNA表达分别为0.7500±0.0038、0.660 0±0.005 5,表达下降（P＜0.05）.0.10μmol/L浓度组作用24、48 h,Caspase-3 mRNA表达分别为4.30±0.29、6.79±0.13,表达上升（P＜0.05）.结论 葫芦素B对人结肠腺癌细胞株LS174-T的增殖具有抑制作用.

Objective To investigate the inhibitory effect of cucurbitacin B on proliferation of human colon cancer cells.Methods LS174-T cells were treated with different concentrations of cucurbitacin B （0.01,0.10,1.00,10.00,and 100.00 mol/L） for 24,48 and 72 h.The cell proliferation was detected by cell counting kit-8 （CCK-8） method,and the distribution of cell cycle by flow cytometry.signaling transducers and activators of transcription 3 （STAT3）,Caspase-3,and Tert mRNA were detected by fluorescence quantitative polymerase chain reaction （FQ-PCR）.Results CCK-8 results showed that after treatment with cucurbitacin B （0.01 and 100.00 moL/L） for 24 h,inhibitory rate of LS174-T cell proliferation was （5.22 ±2.02）% and （73.90 ± 1.85）% respectively.After treatment with 100.00 mol/L of cucurbitacin B for 24 h and 48 h,inhibitory rate was （73.90 ± 1.85）% and （98.84 ±0.51）% respectively.The statistically significant difference was dose-and time-dependent （P 〈 0.05）.Flow cytometry revealed that as compared with control group,the percentage of cells in G2/M phase in 1.00 μmol/L concentration group was increased [（6.33 ± 0.74） % and （30.43 ± 4.09） %],and that in G0/G1 phase decreased [（78.5 ± 4.72） % and （43.2 ± 5.65） %] with the difference being statistically significant （P 〈0.05）.The FQ-PCR indicated that after treatment with cucurbitacin B of 0.01 and 10.00 mol/L for 24 h,STAT3 level was 0.750 0 ±0.003 8,0.660 0 ±0.005 5,respectively.After treatment with cucurbitacin B of 0.10 μmol/L for 24 h and 48 h,Caspase-3 mRNA level was 4.30 ±0.29 and 6.79 ±0.13 respectively with the difference being statistically significant （P 〈 0.05）.Conclusion Cucurbitacin B can inhibit proliferation of colon cancer cells.