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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 of Janus kinase inhibitor AG490 on the proliferation, apoptosis and radiosensitivity of esophageal squamous cell carcinoma Eca-109 cells and to explore its mechanism. Methods Eca-109 cells were treated with different concentrations of AG490 (0, 25, 50 and 100 μmol / L). Cell proliferation was detected by MTT assay. Apoptosis was detected by flow cytometry with Annexin V-FITC / Cell radiosensitivity and Western blot were used to detect the expression of Stat3, p-Stat3, Bcl-2 and Bax in each group. Results Compared with the control group, the proliferation of Eca-109 cells on the 1st to 5th day after AG490 treatment was dose-response-dependent (P <0.05, P <0.01) in AG490 concentration groups (25,50,100μmol / L) ). Compared with the control group, the apoptotic rates of Eca-109 cells in AG490 concentration groups (25, 50 and 100μmol / L) were increased after AG490 treatment for 48 h (P <0.01). Compared with the control group, the survival rate of the cells treated with 50μmol / L X-ray irradiation decreased, the biological parameters SF2, D0, Dq and N decreased (P <0.05) and the SERD0 and SERDq increased (P <0.05) . Compared with the control group, the expressions of p-Stat3 protein in AG49050μmol / L + 4Gy group and AG49050μmol / L group were down-regulated (P <0.05, P <0.01). Compared with the control group, the expression of Bcl-2 was decreased (P <0.05) and the expression of Bax was increased (P <0.05) at the concentrations of 50 and 100μmol / L in AG490. Conclusion AG490 inhibits the proliferation, increases the apoptosis of Eca-109 cells and increases the sensitivity of radiotherapy. The possible mechanism is that AG490 regulates the expression of Bcl-2 and Bax by blocking the activation of Stat3.