Regulation of THZ1,an inhibitor of cyclin-dependent kinase 7,on stemness of glioma stem cells and its mechanism
- VernacularTitle:细胞周期蛋白依赖性激酶7抑制剂THZ1对脑胶质瘤干细胞干性的调控及机制
- Author:
Enxi HU
1
;
Wenying HE
;
Xiang TAO
;
Peijing DU
;
Libin WANG
Author Information
- Publication Type:Journal Article
- Keywords: THZ1; glioma; cancer stem cell; mammosphere culture; stemness marker; Wnt/β-catenin signaling pathway; epithelial-mesenchymal transformation; proliferation; migration
- From: Chinese Journal of Tissue Engineering Research 2025;29(25):5374-5381
- CountryChina
- Language:Chinese
- Abstract: BACKGROUND:THZ1,an inhibitor of cyclin-dependent kinase 7,has been shown to inhibit the proliferation of a variety of tumor cells,but whether THZ1 can affect the stemness of glioma stem cells through the Wnt/β-catenin signaling pathway remains unclear.OBJECTIVE:To investigate the effect of THZ1 on stemness of glioma cell U87 and its mechanism.METHODS:U87 adherent cells were cultured to form stem cell mammospheres.The expressions of stemness related proteins were verified by western blot assay.The effect of THZ1 on half maximal inhibitory concentration(IC50)of U87 cells was determined by Cell Counting Kit-8(CCK-8)assays.The effects of THZ1 on proliferation and migration of U87 cells were determined by cell colony-formation assays,cell wound healing assays,and Transwell migration assays.The effect of THZ1 treatment on mammosphere forming rate and mammosphere size of U87 stem cells was analyzed.Stemness associated proteins CD133,ABCG2,Nanog,OCT4,SOX2,epithelial-mesenchymal transformation-related proteins E-cadherin,N-cadherin,Occludin,Snail,and Wnt/β-catenin pathway associated proteins Axin1,β-Catenin,WNT-5A,GSK3β,Cyclind-1,and C-myc were measured by western blot assay.RESULTS AND CONCLUSION:(1)Compared with adherent cells,the expressions of stemness related proteins Nestin,CD133,ABCG2,Nanog,OCT4,and SOX2 were significantly increased.(2)Compared with the control group,THZ1 decreased the proliferation and migration of U87 cells.(3)THZ1 inhibited the mammosphere forming rate and mammosphere size of U87 stem cells.(4)After THZ1 treatment,the expression of N-cadherin and Snail decreased,while the protein expression of E-cadherin and Occludin increased.(5)THZ1 treatment decreased the expression of Wnt/β-catenin pathway related proteins Axin1,β-Catenin,Wnt-5A,GSK3β,Cyclind-1,and C-myc in U87 stem cells.It is concluded that THZ1 can suppress the proliferation and migration of U87 cells,and inhibit the mammosphere forming ability,stemness related protein expression,and epithelial-mesenchymal transformation ability of U87 stem cells by down-regulating the expression of Wnt/β-catenin signaling pathway related molecules.
