Pharmacologic targeting of the P-TEFb complex as a therapeutic strategy for chronic myeloid leukemia.
Pharmacologic targeting of the P-TEFb complex as a therapeutic strategy for chronic myeloid leukemia.
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P-TEFb 复合物的药理学靶向作为慢性粒细胞白血病的治疗策略
DOI：
10.1186/s12964-021-00764-5
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发表时间：
2021-08-09
期刊：
影响因子：
--
通讯作者：
Hui H
中科院分区：
文献类型：
--
作者：
Qing Y;Wang X;Wang H;Hu P;Li H;Yu X;Zhu M;Wang Z;Zhu Y;Xu J;Guo Q;Hui H
背景正转录延伸因子b (P-TEFb) 激酶活性参与转录过程。细胞周期蛋白依赖性激酶 9 (CDK9) 是 P-TEFb 的核心成分，调节转录延伸过程，这与许多癌症类型的分化和细胞凋亡相关。汉黄芩素是从黄芩中分离得到的天然CDK9抑制剂。本研究旨在探讨汉黄芩素抗慢性粒细胞白血病(CML)细胞的分子机制。材料与方法采用RT-qPCR和Western blot分析mRNA和蛋白水平。流式细胞术用于评估细胞分化和凋亡。应用细胞转染、免疫荧光分析和免疫共沉淀（co-IP）测定来研究汉黄芩素的潜在调节机制。采用KU-812细胞异种移植NOD/SCID小鼠模型来评估和验证体内机制。结果我们报道汉黄芩素诱导的K562、KU-812和原代CML细胞的抗CML作用是由P-TEFb复合物调节的。我们还通过敲低CDK9的表达证实了CDK9与红系分化之间的关系。为进一步研究汉黄芩素诱导红系分化的机制，采用co-IP实验证明汉黄芩素可增加GATA-1与FOG-1之间的结合，但降低GATA-1与RUNX1之间的结合，而这种结合依赖于P-TEFb。此外，汉黄芩素还可诱导细胞凋亡并降低 KU-812 细胞中 MCL-1（P-TEFb 的下游）的 mRNA 和蛋白水平。体内研究表明汉黄芩素在KU-812异种移植NOD/SCID小鼠模型中具有良好的抗肿瘤作用，并降低小鼠脾脏中人CD45+细胞的比例。我们还验证了汉黄芩素在体内通过调节P-TEFb活性发挥抗CML作用。结论我们的研究表明了CML细胞中P-TEFb激酶活性调节的特殊机制，为汉黄芩素在CML临床治疗中的进一步应用提供了依据。视频摘要
BackgroundThe positive transcription elongation factor b (P-TEFb) kinase activity is involved in the process of transcription. Cyclin-dependent kinase 9 (CDK9), a core component of P-TEFb, regulates the process of transcription elongation, which is associated with differentiation and apoptosis in many cancer types. Wogonin, a natural CDK9 inhibitor isolated fromScutellaria baicalensis.This study aimed to investigate the involved molecular mechanisms of wogonin on anti- chronic myeloid leukemia (CML) cells.Materials and methodsmRNA and protein levels were analysed by RT-qPCR and western blot. Flow cytometry was used to assess cell differentiation and apoptosis. Cell transfection, immunofluorescence analysis and co-immunoprecipitation (co-IP) assays were applied to address the potential regulatory mechanism of wogonin. KU-812 cells xenograftNOD/SCIDmice model was used to assess and verify the mechanism in vivo.ResultsWe reported that the anti-CML effects in K562, KU-812 and primary CML cells induced by wogonin were regulated by P-TEFb complex. We also confirmed the relationship between CDK9 and erythroid differentiation via knockdown the expression of CDK9. For further study the mechanism of erythroid differentiation induced by wogonin, co-IP experiments were used to demonstrate that wogonin increased the binding between GATA-1 and FOG-1 but decreased the binding between GATA-1 and RUNX1, which were depended on P-TEFb. Also, wogonin induced apoptosis and decreased the mRNA and protein levels of MCL-1 in KU-812 cells, which is the downstream of P-TEFb. In vivo studies showed wogonin had good anti-tumor effects in KU-812 xenograftsNOD/ SCIDmice model and decreased the proportion of human CD45+cells in spleens of mice. We also verified that wogonin exhibited anti-CML effects through modulating P-TEFb activity in vivo.ConclusionsOur study indicated a special mechanism involving the regulation of P-TEFb kinase activity in CML cells, providing evidences for further application of wogonin in CML clinical treatment.Video Abstract
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11.4
作者：
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