吉林大学学报(医学版)

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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

李 翔,张 斌,马 竟,高 琦,史 毅   

  1. 辽宁医学院附属第一医院口腔科,辽宁 锦州 121001
  • 收稿日期:2013-11-16 出版日期:2014-05-28 发布日期:2014-06-05
  • 通讯作者: 张 斌(Tel:0416-4197029,E-mail:5100973@qq.com) E-mail:5100973@qq.com
  • 作者简介:李 翔(1987-),女,新疆维吾尔自治区乌鲁木齐市人,在读医学硕士,主 要从事舌癌细胞生物学的研究。 
  • 基金资助:

    辽宁省教育厅高等学校科学研究项目资助课题(L2010315)

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

LI Xiang,ZHANG Bin,MA Jing,GAO Qi,SHI Yi   

  1. Department of Stomatology,First Affiliated Hospital,Liaoning Medical University,Jinzhou 121001,China
  • Received:2013-11-16 Online:2014-05-28 Published:2014-06-05

摘要:

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250 nmol/L 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36 h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。FCM法检测,1、5、25、125和250 nmol?L-1 MK2206作用舌鳞癌TCA-8113细胞12 h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。

关键词:  , 舌鳞状细胞癌, 细胞增殖, 细胞凋亡, PI3K/Akt通路

Abstract:

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 MK2206 groups.The inhibitory rate of proliferation of TCA-8113 cells was detected with MTT method,and the a
poptotic 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.The FCM results showed that the apoptotic rates of TCA-8113 cells after treated with 1,5,25,125, and 250 nmol/L 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-3β were decreased with the increasingof dose and time of MK2206; compared with  the  β-actin in  control group,the bands got darken;the expression level of caspase-9 was increased,compared with the β-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 squamou
s cell carcinoma TCA-8113 cells and induce apoptosis.

Key words: tongue squamous cell carcinoma; cell proliferation, apoptosis, PI3K/Akt pathway

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