AG490对胆囊癌细胞JAK2/STAT3信号传导通路的抑制作用研究

Inhibitory effect of AG490 on JAK2/STAT3 signal pathway in gallbladder carcinoma cells

目的研究JAK激酶抑制剂AG490对人胆囊癌细胞株GBC-SD生长增殖的影响,探讨AG490对JAK2/STAT3信号传导通路的抑制作用及其对胆囊癌治疗的意义。方法培养GBC-SD细胞,根据研究目的设置实验组和对照组,在第一项实验中,实验组以3个浓度梯度的AG490作用于GBC-SD细胞,对照组加入等量PBS,继续培养,24 h后以MTT法测定OD值,反映各组细胞的增殖活性;在第二项实验中,实验组以100μmol/mol的AG490作用24 h,对照组平行培养,收集两组细胞进行免疫印迹法检测,观察AG490对细胞中JAK2、STAT3蛋白表达的影响。结果 AG490能有效抑制胆囊癌细胞增殖,且抑制作用呈浓度依赖性,即浓度越高,抑制作用越明显,不同浓度组细胞增殖抑制率差异有统计学意义(P<0.05);AG490作用后细胞中JAK2、STAT3蛋白浓度降低,与对照组比较差异亦有统计学意义(P<0.05)。结论 AG490可以抑制细胞中JAK2、STAT3蛋白的表达,通过抑制JAK2/STAT3信号传导通路发挥抑制胆囊癌细胞增殖的作用。

Objective To study the impact of JAK kinase inhibitor AG490 on the growth of human gallbladder carcinoma cell GBC-SD, and to investigate the inhibitory effect of AG490 on JAK2/STAT3 signaling pathway and its implications for the treatment of gallbladder cancer. Methods GBC-SD cell was cultivated and grouped into testing group and control group according to this project purpose. In the first part, 3 different concentrations of AG490 was used on GBCSD cells of test group, while the control group was cultured normally PBS, MTT assay was used for testing cell proliferation OD after 24 hours to see its activity of proliferation. On the second part of experiment, the cells in testing group endured 100 μmol/mol of AG490 for 24 hours, after which cells in both groups were collected and handled, later, Western blot was performed to detect the impact of AG490 on the expression level of JAK and STAT3 protein. Results AG490 could inhibit cell proliferation in a concentration-dependent manner, the higher concentration resulted in higher inhibition, namely the inhibition rate of cell proliferation between different concentrations of groups was different, and the difference was statistically significant （P〈0.05）. Another founding that JAK2 and STAT3 protein both reduced compared with the control group with statistically significant difference （P〈0.05）. Conclusion AG490 can inhibit the proliferation of gallbladder cancer cells by inhibiting the expression of JAK2 and STAT3 protein, and it can interfere the JAK2/STAT3 signal pathway to perform its inhibition effect.