补骨脂乙素对舌鳞状细胞癌Tca8113细胞增殖的抑制作用和凋亡诱导作用及其机制

Effects of isobavachalcone on proliferation and apoptosis of tongue squamous cell carcinoma Tca-8113 cells and their mechanisms

目的:观察补骨脂乙素(IBC)对人舌鳞状细胞癌Tca8113细胞增殖和凋亡的影响,并阐明其可能的作用机制。方法:将体外培养的Tca8113细胞分为对照组(0μmol·L-1)和不同浓度IBC组(10、20、40及80μmol·L-1)。通过MTT法检测细胞增殖抑制率,流式细胞术检测细胞凋亡率,Western blotting法检测各组细胞中Akt、p-Akt、Erk、p-Erk、Bax、Bcl-2和Caspase-3蛋白的表达水平。结果:MTT法检测,随着IBC浓度的增加和作用时间的延长,Tca8113细胞的增殖抑制率逐渐增加,12、24和48h半数抑制浓度(IC50)分别为(285.13±8.97)、(132.40±7.76)和(58.56±5.93)μmol·L-1,不同时间点IC50比较差异有统计学意义(P<0.05)。流式细胞术检测,与对照组(1.69%±0.65%)比较,作用48h后20和40μmol·L-1 IBC组细胞凋亡率(分别为8.21%±2.32%和22.45%±1.18%)明显升高(P<0.05)。Western blotting法检测,与对照组比较,作用48h后20和40μmol·L-1 IBC组细胞中Bcl-2、p-Akt和p-Erk表达水平明显降低(P<0.05),Bax表达水平明显升高(P<0.05);Akt和Erk蛋白表达水平无明显变化(P>0.05);与对照组比较,作用48h后40μmol·L-1 IBC组Tca8113细胞中Caspase-3蛋白表达水平明显升高(P<0.05)。结论:IBC可抑制Tca8113细胞的增殖并诱导细胞凋亡,Akt和Erk通路可能是其诱导肿瘤细胞凋亡的途径。

Objective：To observe the effects of isobavachalcone（IBC）on the proliferation and apoptosis of tongue squamous cell carcinoma Tca-8113 cells,and to explore their mechanisms.Methods：The Tca8113 cells cultured in vitro were divided into control group and different doses（0,10,20,40,80μmol·L^-1）of IBC groups.The inhibitory rates of cell proliferation were detected by MTT method.The apoptosis was detected by flow cytometry.Western blotting method was used to detect the expressions of Akt,p-Akt,Erk,p-Erk,Bax,Bcl-2and Caspase-3proteins in the Tca8113 cells in various groups.Results：The MTT results showed that the inhibitory rates of proliferation of Tca8113 cells were increased in a concentration-and time-dependent manner;the IC50 at 12,24 and 48hwere（285.13±8.97）,（132.40±7.76）,and（58.56±5.93）μmol·L^-1,respectively;and there were significant differences between different time points（P〈0.05）.The FCM results showed that the apoptotic rates of Tca8113 cells in 20 and 40μmol·L^-1 IBC groups at 48hwere（8.21±2.32）% and（22.45±1.18）%,respectively;compared with control group（1.69% ± 0.65%）,the differences were statistically significant（P〈0.05）.The Western blotting results showed that the expression levels of Bcl-2,p-Akt,and p-Erk,in 20 and 40μmol·L^-1 IBC groups were decreased and the expression levels of Bax were increased compared with control group（P〈0.05）;the expression levels of Akt and Erk had no significant changes（P〈0.05）.The expression levels of Caspase-3in Tca8113 cells in 40μmol·L^-1 IBC group at 48 hwere increased compared with control group（P〈0.05）.Conclusion：IBC could inhibit the proliferation and induce the apoptosis of Tca8113cells;Akt and Erk signaling pathway may be the pathway of IBC to induce the apoptosis of tumor cells.