AG490对食管鳞癌Eca-109细胞增殖、凋亡及放疗敏感性的影响  

Effect of Janus kinase inhibitor AG490 on proliferation,apoptosis and radiosensitivity of human esophageal carcinoma Eca-109 cells

作　　者：臧春宝[1] 万跃[2] 王勇[1] 侯妍利[1] 唐敏[1] 李少林[1] 

机构地区：[1]重庆医科大学基础医学院放射医学教研室,重庆400016 [2]重庆市肿瘤医院放疗科,重庆400030

出　　处：《第三军医大学学报》2013年第13期1357-1361,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金(81171365)~~

摘　　要：目的研究Janus激酶抑制剂AG490对食管鳞癌Eca-109细胞增殖、凋亡及放疗敏感性的影响,并探讨其作用机制。方法用不同浓度(0、25、50、100μmol/L)的AG490处理Eca-109细胞,采用MTT法检测细胞增殖,AnnexinV-FITC/PI双染流式细胞仪检测细胞凋亡,克隆形成实验检测细胞放疗敏感性,Western blot检测各组细胞Stat3、p-Stat3、Bcl-2、Bax蛋白表达。结果与对照组比较,AG490各浓度组(25、50、100μmol/L)在AG490作用后第1～5天Eca-109细胞增殖减慢,呈量效-时效关系(P<0.05,P<0.01)。与对照组比较,AG490各浓度组(25、50、100μmol/L)在AG490作用48 h后Eca-109细胞凋亡率增加,呈量效关系(P<0.01)。与对照组比较,各剂量X射线照射后50μmol/L组细胞存活率降低,放射生物学参数SF2、D0、Dq、N值均降低(P<0.05),SERD0和SERDq值增加(P<0.05)。与对照组比较,AG49050μmol/L+4Gy组和AG490 50μmol/L组p-Stat3蛋白表达下调(P<0.05,P<0.01)。与对照组比较,AG490各浓度(50和100μmol/L)组Bcl-2的表达降低(P<0.05)、Bax的表达增加(P<0.05)。结论 AG490抑制Eca-109细胞增殖,增加细胞凋亡,增加放疗敏感性,其机制可能是通过阻断Stat3蛋白的活化,调控凋亡蛋白(Bcl-2和Bax)的表达。Objective To investigate the effect and mechanism of Janus kinase inhibitor AG490 on the proliferation, apoptosis and radiosensitivity of human esophageal carcinoma Eca-109 cells. Methods Eca-109 ceils were treated with different concentrations of AG490 （0, 25, 50 and 100 p, moL/L）. Cell proliferation and apoptosis were detected by M3F assay and flow cytometry, respectively, and cell radiosensitivity was measured by colony formation assay. The expression levels of Stat3, p-Stat3, Bcl-2 and Bax were evaluated by Western blotting. Results Compared with the control group （0 μmol/L）, the proliferation of Eca-109 cells in AG490 treatment groups （25, 50 and 100 μmol/L） significantly decreased at 1-5 d after treatment in a dose- and time-dependent manner （P 〈0.05, P 〈0.01 ）. Compared with the control group （0 μmol/L）, the apoptosis of Eca-109 cells in AG490 treatment groups （25, 50 and 100 μmoL/L） significantly increased at 48 h after treatment in a dose-dependent manner （P 〈 0.01 ）. Compared with the control group （0 μmol/L）, the cell survival fraction and radiobiological parameters （SF2, Do, Dq and N） of the AG490 50μmol/L group significantly decreased （P 〈 0.05 ）, and the sensitivity enhancement ratio （SERD0 and SERDq ） significantly increased （P 〈 0.05）. Compared with the control group （0 μmol/L）, the expression of p-Star3 in AG490 50 μmol/L ＋ 4 Gy group and AG490 50 μmol/L group significantly decreased （ P 〈 0.05, P 〈 0.01 ）. Compared with the control group （0 μmol/L）, the expression of Bcl-2 in AG490 treatment groups （50 and 100 μmol/L） significantly decreased （ P 〈 0.05 ）, and the expression of Bax siznificantlv increased （ P 〈0. 05 ）. Conclusion AG490 can regulate the expression of Bcl-2 and Bax by blocking the activation of Stat3, thus inhibiting the proliferation and increasing the apoptosis and radiosensitivity of human esophageal carcinoma Eca-109 cells in vitro.

关 键 词：AG490 食管鳞癌 STAT3 放疗敏感性 

分 类 号：R73-36[医药卫生—肿瘤]