论文部分内容阅读
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: AG490, an inhibitor of JAK2 / STAT3 pathway, is expected to be a potential drug in the clinical treatment of tumors because of its high efficiency and low toxicity in inhibiting tumor cells. However, the anti-tumor mechanism of AG490 is not yet clear. Therefore, this article intends to AG490 inhibition of lymphoma cell proliferation and its mechanism of action to further explore for AG490 clinical application to provide experimental evidence. METHODS: Lymphoma cells (Namalwa and JeKo-1), Jurkat T lymphocytic leukemia cells and THP-1 monocytic leukemia cells were treated with different doses of AG490 for 24 hours. AG490 (0μM, 2μM, 20μM, 50μM, 200μM). The changes of BATF2 mRNA were detected by real-time quantitative PCR. The protein level was detected by Western blot. The expression of BATF2 was inhibited by siRNA transfected into cells. CCK8 assay was used to detect the effect of AG490 on Namalwa cells Proliferation inhibitory effect. Results: AG490 inhibited the proliferation of Namalwa, JeKo-1 and Jurkat cells in a dose-dependent manner (P <0.05), and up-regulated the expression of BATF2 mRNA and protein (P <0.05). BATF2 expression also did not increase in THP-1 cells without significant inhibition (P> 0.05). The inhibitory effect of AG490 on the proliferation of Namalwa cells was significantly reduced by siRNA inhibition of BATF2 gene expression (P <0.05). CONCLUSION: The efficiency of AG490 killing tumor cells is positively correlated with the expression of BATF2 induced by AG490. The inhibitory effect of AG490 on the proliferation of tumor cells with BATF2 expression is significantly reduced. Therefore, AG490 may inhibit lymphoma cell proliferation by up-regulating BATF2 expression. This means that BATF2 is the target of AG490 killer lymphoma cells and may contribute to the development of new drugs.