α桐酸对舌鳞状细胞癌CAL-27细胞的体外抑制作用

Inhibitory effect ofα-eleostearic acid on tongue squamous cell carcinoma CAL-27

目的:探讨苦瓜籽油中的α桐酸(α-eleostearic acid,α-ESA)对舌鳞状细胞癌CAL-27细胞增殖、迁移、凋亡的影响。方法:将0、70、80、90、100、110、120μmol/L的α-ESA培养CAL-27细胞后,用CCK-8法和流式细胞术分别检测加药24、48、72 h后细胞增殖抑制率和加药48 h后细胞凋亡率。再分别以0、80、100μmol/L的α-ESA培养CAL-27细胞不同时间后,用细胞划痕实验检测加药24 h后细胞的迁移能力,Hoechst 33258染色在荧光显微镜下观察加药48 h后凋亡细胞形态。采用SPSS 21.0软件包对实验数据进行统计学分析。结果:α-ESA对CAL-27细胞的增殖抑制作用呈明显时间和浓度依赖性(P<0.05)。作用24、48、72 h后,半数抑制浓度(IC50)逐渐降低,分别为(123.48±1.00)、(80.22±0.03)和(69.27±80.34)μmol/L。流式细胞检测结果显示,细胞凋亡率随着α-ESA浓度的增加而大幅增高(P<0.05)。培养24 h后,加入80、100μmol/L的α-ESA的细胞划痕愈合率分别为(40.08±2.19)%、(22.06±2.04)%,显著低于对照组(74.74±2.17)%(P<0.01)。荧光显微镜下观察到致密浓染或碎块状亮蓝色荧光的细胞凋亡现象。结论:α-ESA能够抑制CAL-27细胞的体外增殖及迁移能力,并可诱导细胞凋亡,从而产生一定的抗肿瘤生长作用。

PURPOSE:To investigate the effect ofα-eleostearic acid(α-ESA)in bitter melon seed oil on the proliferation,migration and apoptosis of tongue squamous cell carcinoma CAL-27 cells.METHODS:CAL-27 cells were cultivated in vitro andα-ESA was added at the concentrations of 0,70,80,90,100,110 and 120μmol/L.CCK-8 assay and flow cytometry were used to detect the cell proliferation inhibition rate at 24,48,and 72 h and apoptosis rate at 48 h respectively.CAL-27 cells were treated withα-ESA at concentrations of 0,80 and 100μmol/L.The cell migration capacity after 24 hours ofα-ESA administration was measured by cell scratching assay.The morphology of apoptotic cells after 48 h ofα-ESA administration was observed by Hoechst 33258 staining.SPSS 21.0 software package was used for statistical analysis.RESULTS:The proliferation of CAL-27 cells was inhibited byα-ESA in a time and concentrationdependent manner(P<0.05).After 24,48,and 72 h ofα-ESA administration,the half inhibitory concentration(IC50)was(123.48±1.00)μmol/L,(80.22±0.03)μmol/L,and(69.27±80.34)μmol/L,respectively,which was gradually decreased.The results of flow cytometry showed that the apoptosis rate increased significantly with the increase ofα-ESA concentration(P<0.05).After 24 h of incubation,the wound healing rate of CAL-27 cells was(40.08±2.19)%and(22.06±2.04)%after administration of 80 and 100μmol/Lα-ESA,which was significantly lower than the control group(74.74±2.17)%(P<0.01).Densely stained or fragmented bright blue fluorescence was observed under fluorescence microscope which represented cell apoptosis.CONCLUSIONS:α-ESA can inhibit the proliferation and migration of CAL-27 cells in vitro,and therefore has certain anti-tumor effects.