白桦脂酸抑制食管鳞状细胞癌增殖及相关机制的实验研究

 目的研究抗肿瘤药物白桦脂酸（BA）对人食管鳞癌KYSE170细胞的抑制作用及抗肿瘤作用机制。方法采用不同质量浓度(0、5、10、20、40、60、80、100 μg·mL-1)白桦脂酸处理KYSE170细胞，作用不同时间 (24、48、72 h)后 ，噻唑蓝（MTT）法检测KYSE170 细胞生长的短期抑制作用；不同质量浓度（0、5、20、40 μg·mL-1）白桦脂酸作用于细胞24 h后以克隆形成实验检测药物对细胞生长的长期抑制作用；不同质量浓度白桦脂酸 (10、60、100 μg·mL-1)作用KYSE170细胞 24、48 h后以流式细胞仪FITC Annexin V/PI双染法检测细胞凋亡变化，不同质量浓度白桦脂酸（0、10、40 μg·mL-1）作用细胞24 h后以流式细胞仪检测细胞周期的改变。结果噻唑蓝实验显示，不同质量浓度白桦脂酸对KYSE170细胞均有抑制作用，抑制效果呈时间和剂量依赖性，24、48、72 h的IC50分别为(5681±256)、(3973±277)和(2928±305)  μg·mL-1；克隆形成结果显示，0、5、20、40 μg·mL-1药物作用细胞后，克隆形成率分别为（8956+500）%、（6100+203）%、（3133+351）%和（1533+233）%，随白桦脂酸质量浓度增大而明显降低（P< 001）。凋亡结果显示， 各实验组KYSE170细胞凋亡率随白桦脂酸质量浓度加大和药物作用时间的延长明显增高（P＜ 005或001）。细胞周期结果显示，0、1、10 μg·mL-1白桦脂酸作用细胞24 h后，随药物质量浓度增大，G1期细胞比例减少，S期细胞比例不断增加，实验组与空白组对比均有统计学差异（P<001）。结论白桦脂酸能抑制人食管鳞癌KYSE170细胞的生长，这一作用可能是通过诱导细胞凋亡、阻滞细胞停留在S期来实现的。

Inhibitory Effect and Mechanisms of Betulinc Acid on Esophageal Squamous Carcinoma Cell

OBJECTIVE To explore the inhibitory effect of betulinc acid(BA) on esophageal squamous cell carcinoma(ESCC) KYSE170 cells and the mechanisms of the inhibitory effect.METHODS Different concentrations of BA(0,5,10,20,40,60,80 and 100 μg·mL-1) were used to detect their effects and the inhibitory rate on the cells for 24 to 72 h.The clone formation test was used to detect the long term inhibitory effects of different BA concentration(0,5,20 and 40 μg·mL-1)on KYSE170 cells.Different concentrations of BA(10,60 and 100 μg·mL-1) for 24 and 48 h were used to detect the apoptosis rate of cells by flow cytometry.Different concentrations of BA(0,10 and 40 μg·mL-1) for 24 h were used to detect the cell cycle of cells by flow cytometry.RESULTS BA inhibited the growth of KYSE170 cells in a dose-and time-depentent manner.IC50 at 24,48 and 72 h were(56.81±2.56),(39.73±2.77) and(29.28±3.05) μg·mL-1,respectively.The clone formation plating efficiencies were(89.56+5.00)%,(61.00±2.03)%,(31.33±3.51)% and(15.33±2.33)% when the drug concentrations were 0,5,20 and 40 μg·mL-1,respectively.With the increase of BA concentration and the prolong of BA incubation time the apoptosis rate increased significantly(P< 0.05 or 0.01).After 0,1 and 10 μg·mL-1BA added for 24 h,the rate of G1 phase cells decreased,while the rate of S phase cells increased(P<0.01).CONCLUSION BA can inhibit the growth of KYSE170 cell by inducing cell apoptosis and blocking cells to stay in S phase.