AG490上调BATF2表达抑制淋巴瘤细胞增殖

AG490 Suppresses Lymphoma Cell Proliferation by up-regulating BATF2 Expression

目的:AG490作为JAK2/STAT3通路的抑制剂,在对肿瘤细胞的抑制作用上所展现出的高效低毒性,使其有望成为临床上治疗肿瘤的一种可能的药物。然而,AG490的抗瘤机制尚未明确。因此,本文拟对AG490抑制淋巴瘤细胞增殖的效应及其作用机制进行进一步探讨,为AG490应用于临床提供实验依据。方法:用不同剂量的AG490处理淋巴瘤细胞(Namalwa和JeKo-1)、Jurkat T淋巴细胞性白血病细胞和THP-1单核细胞性白血病细胞24小时,CCK-8法检测AG490(0μM、2μM、20μM、50μM、200μM)对上述细胞的增殖抑制作用,实时定量PCR法检测BATF2 mRNA的变化,Western blot法检测其蛋白水平的变化,细胞转染siRNA法抑制BATF2表达后CCK8法检测AG490对Namalwa细胞的增殖抑制效应。结果:AG490呈剂量依赖性地抑制Namalwa、JeKo-1、Jurkat细胞的增殖(P<0.05),同时上调其BATF2 mRNA水平和蛋白水平的表达(P<0.05)。对于无显著抑制作用的THP-1细胞,BATF2的表达亦未见升高(P>0.05)。siRNA法抑制BATF2基因表达后,AG490对Namalwa细胞的增殖抑制效果明显降低(P<0.05)。结论:AG490杀肿瘤细胞的效率与其诱导的BATF2的表达呈正相关,抑制BATF2的表达后AG490抑制肿瘤细胞增殖的效率明显降低。因此,AG490可能是通过上调BATF2表达的方式抑制淋巴瘤细胞增殖。这意味着BATF2是AG490杀伤淋巴瘤细胞的作用靶点,可能为新药的开发做出一定的贡献。

Objective： The inhibitory effect of AG490, the JAK2/STAT3 pathway inhibitor, on tumor cells with high efficiency and low toxicity makes it a potential drug that is clinically used to treat tumor. Nevertheless, the mechanism how AG490 suppresses tumor remains not elusive. Therefore, this paper will investigate the effect and the mechanism of AG490 suppressing lymphoma cell proliferation. Methods： Lymphoma cells （Namalwa and JeKo-1）, T-cell leukemia cells（Jurkat） and monocytic leukemia cells（THP-1） were treated with different doses ofAG490（0 μM, 20 μM and 50 μM） for 24h. Cell viability was measured by CCK-8 assay. The mRNA level of BATF2 was detected by real-time PCR, and the protein level was detected by Western blot. Namalwa cells were transfected with siR- NA to inhibit BATF2 expression. Results： Namalwa, JeKo-1 and Jurkat cells were dose-dependently suppressed by AG490 （P〈0.05）. Correspondingly, the mRNA and protein expression of BATF2 were significantly up-regulated （P〈0.05）. However, no significant change was shown on cell proliferation suppression or BATF2 expression of THP-1 cells （P〉0.05）. AG490 couldn＇t inhibit Namalwa cells prolif- eration when BATF2 was repressed by siRNA. Conclusion： The inhibitory effect of AG490 on lymphoma cells was positively correlated with the BATF2 expression induction. Moreover, the inhibitory effect was decreased after BATF2 gene was knocked down. Thus, upreg- ulation of BATF2 may be a possible mechanism, by which AG490 inhibits the proliferation of lymphoma cells. These results imply that BATF2 is the target of AG490 for its anti-lymphoma effect, which might contribute to the development of new drugs.