摘要
目的研究不同孵育时间以及不同浓度的STAT3特异性抑制剂AG490作用于人移形上皮膀胱癌细胞株(pumc-91)中STAT3信号通路蛋白表达的条件优化。方法分别在0、24、36、48、60h收集细胞并提取细胞蛋白,应用Western Blot对各组中STAT3的蛋白表达水平进行半定量分析;加入100μmol/L AG490作用于pumc-91细胞,分别用0、50、100、200μmol/L的AG490处理pumc-91细胞48h后,应用Western Blot检测各组中STAT3的蛋白表达水平。结果 STAT3蛋白的表达随AG490作用时间延长而逐步降低,在作用时间为48h时,差异有统计学意义(P<0.05)。STAT3蛋白的表达明显随AG490浓度的升高而逐步下降,在浓度为200μmol/L时差异具有统计学意义(P<0.05)。结论 AG490通过抑制STAT3蛋白的表达,成功抑制了STAT3信号通路的活性,而且其抑制作用具有时间和剂量的依赖性,AG490在作用浓度为200μmol/L和作用时间为48h的条件下,有效的抑制了STAT3蛋白在pumc-91细胞系中的表达。
Objective To optimize the best condition of the effect of AG490,a STAT3 inhibitor on human transition cell carcinoma of the urinary bladder cell line（ pumc-91）. Methods Pumc-91 cells were given AG490 at 100μmol / L and then cultured separately for 0,24,36,48 and 60 hours. The others were treated with 0,50,100,200μmol / L of AG490 respectively and cultured for 48 h. Western blot was adopted to detect the expression of STAT3. Results With the growth of the action time of AG490,the expression of STAT3 protein decreased gradually. When the action time was 60 h,the difference was statistically significant（ P〈0. 05）. With the increase of the concentration of AG490,the expression of STAT3 protein decreased gradually. In the concentration of 200μmol / L,the difference was statistically significant（ P〈0. 05）. Conclusion By inhibiting the expression of STAT3 protein,AG490 can successfully inhibit the active of STAT3 signaling pathways in a time and dose dependent manner. When the concentration is 200μmol / L and the action time is 48 h,AG490 can inhibit the expression of STAT3 protein in pumc-91 cell effectively.
出处
《标记免疫分析与临床》
CAS
2016年第4期451-454,共4页
Labeled Immunoassays and Clinical Medicine
基金
尿液细胞分子诊断北京市重点实验室(Z151100001615060)
北京市中医药管理局(2014-ZYJ04)
关键词
膀胱癌
STAT3
AG490
蛋白免疫印迹
Bladder cancer
Signal transducer and activator of transcription 3（STAT3）
AG490
Western blotting
