1. Academic Validation
  2. 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

  • Cell Commun Signal. 2021 Aug 9;19(1):83. doi: 10.1186/s12964-021-00764-5.
Yingjie Qing  # 1 Xiangyuan Wang  # 1 Hongzheng Wang 1 Po Hu 1 Hui Li 1 Xiaoxuan Yu 2 Mengyuan Zhu 1 Zhanyu Wang 1 Yu Zhu 3 Jingyan Xu 4 Qinglong Guo 5 Hui Hui 6
Affiliations

Affiliations

  • 1 State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, 210009, People's Republic of China.
  • 2 Department of Pharmacology, School of Medicine and Holostic Integrative Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, People's Republic of China.
  • 3 Department of Hematology, The First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing, 210029, People's Republic of China.
  • 4 Department of Hematology, The Affiliated DrumTower Hospital of Nanjing University Medical School, Nanjing, 210008, People's Republic of China.
  • 5 State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, 210009, People's Republic of China. anticancer_drug@163.com.
  • 6 State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing, 210009, People's Republic of China. moyehh@163.com.
  • # Contributed equally.
Abstract

Background: The 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 from Scutellaria baicalensis. This study aimed to investigate the involved molecular mechanisms of wogonin on anti- chronic myeloid leukemia (CML) cells.

Materials and methods: mRNA 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 xenograft NOD/SCID mice model was used to assess and verify the mechanism in vivo.

Results: We 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 xenografts NOD/ SCID mice 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.

Conclusions: Our 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.

Keywords

CDK9; CML; Cell differentiation; P-TEFb; Wogonin.

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