Akt抑制剂MK2206对舌鳞癌TCA-8113细胞增殖和凋亡的影响及其作用机制

Influence of Akt inhibitor MK2206 in proliferation and apoptosis of tongue squamous cell carcinoma TCA-8113 cells and its mechanism

目的:探讨Akt抑制剂MK2206对舌鳞状细胞癌(舌鳞癌)TCA-8113细胞增殖和凋亡的影响,并阐明其可能的作用机制。方法:取处于对数生长期舌鳞癌TCA-8113细胞株随机分为对照组和1、5、25、125及250nmol·L-1 MK2206实验组。在不同剂量及时间处理因素下,采用MTT法检测细胞增殖抑制率,流式细胞术(FCM)检测细胞凋亡率,蛋白质印迹分析检测细胞中caspase-9、Bad、GSK-3β、p-Akt和T-Akt蛋白表达水平。结果:舌鳞癌TCA-8113细胞在MK2206作用12、24和36h的半数抑制浓度(IC50)分别为(112.54±1.67)、(79.67±2.01)和(33.33±1.98)nmol·L-1。FCM法检测,1、5、25、125和250nmol·L-1 MK2206作用舌鳞癌TCA-8113细胞12h后,细胞凋亡率分别为(14.2±0.74)%、(19.3±0.45)%、(35.1±0.45)%、(39.6±0.48)%和(52.1±0.19)%,与对照组比较差异均有统计学意义(P<0.01)。蛋白质印迹分析,随着MK2206药物剂量和作用时间的增加,p-Akt、Bad和GSK-3β蛋白表达水平下降,与对照组β-actin比较其条带的颜色变暗;caspase-9蛋白表达水平升高,与对照组β-actin比较其条带的颜色变深。T-Akt蛋白表达变化不明显,与对照组β-actin比较条带的颜色无明显不同。结论:Akt抑制剂MK2206可抑制舌鳞癌TCA-8113细胞增殖并诱导凋亡。

Objective To explore the influence of Akt inhibitor MK2206 in the proliferation and apoptosis of tongue squamous cell carcinoma TCA-8113 cells,and to clarify the possible mechanism.Methods The tongue squamous carcinoma TCA-8113 cells at the logarithmic phase were randomly divided into control group and 1,5,25,125, 250 nmol·L-1 MK2206 groups.The inhibitory rate of proliferation of TCA-8113 cells was detected with MTT method,and the apoptotic rate of TCA-8113 cells was determined with flow cytometry（FCM）,and the expressions of caspase-9,Bad,GSK-3β,p-Akt and T-Akt proteins in the TCA-8113 cells were detected with Western blotting method.Results The IC50 of tongue squamous cell carcinoma TCA-8113 cells after treated with MK2206 for 12, 24,and 36 h were （112.54±1.67）,（79.67±2.01）,and （33.33±1.98）nmol·L-1 .The FCM results showed＆amp;nbsp;that the apoptotic rates of TCA-8113 cells after treated with 1,5,25,125,and 250 nmol·L-1 MK2206 for 12 h were （14.2±0.74）%,（19.3±0.45）%,（35.1±0.45）%,（39.6±0.48）% and （52.1±0.19）%;there were significant differences compared with control group（P〈0.01）.The Western blotting method results showed that the expressions of p-Akt, Bad and GSK-3βwere decreased with the increasing of dose and time of MK2206;compared with theβ-actin in control group,the bands got darken;the expression level of caspase-9 was increased, compared with theβ-actin in control group, the bands got darken;the T-Akt protein expression did not change significantly;compared with the β-actin in control group, the color of bands had no significant difference.Conclusion Akt inhibitor MK2206 can inhibit the proliferation of tongue squamous cell carcinoma TCA-8113 cells and induce apoptosis.