Protein kinase D inhibitor CRT0066101 suppresses tumor growth by inhibiting the PI3K/AKT signaling pathway to promote apoptosis and autophagy in hepatocellular carcinoma cells
10.12360/CPB202504027
- VernacularTitle:蛋白激酶D抑制剂CRT0066101通过抑制PI3K/AKT信号通路促进肝癌细胞凋亡和自噬抑制肿瘤
- Author:
Hao-hua DENG
1
;
Bao-yuan TANG
;
Bei XIE
;
Lin-jing LI
Author Information
1. 兰州大学第二医院,甘肃兰州 730000
- Publication Type:Journal Article
- Keywords:
hepatocellular carcinoma;
CRT0066101;
autophagy;
apoptosis;
inhibitor;
protein kinase D
- From:
Chinese Pharmacological Bulletin
2025;41(12):2297-2305
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate the inhibitory effect of the protein kinase D(PKD)-specific inhibitor CRT0066101 on hepatocellular carcinoma(HCC)and its underlying molecular mechanisms,providing new theoretical insights and therapeutic strategies for targe-ted HCC treatment.Methods HCC cell lines were treated with varying concentrations of CRT0066101.The inhibitory effects on cell proliferation were assessed using the CCK-8 assay,colony formation assay,and EdU staining.The impact on cell migration and inva-sion was evaluated through wound-healing assays and Transwell migration and invasion assays.was employed toThe effects of CRT0066101 on the phosphorylation levels of PKD and key proteins in the downstream PI3K/AKT signaling pathway were analyzed using Western blot.Additionally,the drug's regulatory effects on apoptosis and autophagy in HCC cells were examined using Western blot,flow cytometry,and the mRFP-GFP-LC3 dual-fluorescence reporter system.Results CRT0066101 significantly inhibited the pro-liferation,migration and invasion of HCC cells.West-ern blotting results demonstrated that CRT0066101 dose-dependently suppressed the phosphorylation of PKD family proteins and downregulated the activation of the PI3K/AKT signaling pathway.Furthermore,CRT0066101 treatment upregulated the expression of the pro-apoptotic protein Bax while downregulating the anti-apoptotic protein Bcl-2.It also markedly increased the expression levels of autophagy marker proteins Bec-lin-1 and LC3B-Ⅱ,suggesting that the drug simulta-neously induced apoptosis and autophagy in HCC cells.Conclusions CRT0066101 specifically inhibits PKD activity,blocks the PI3K/AKT signaling path-way,suppresses HCC cell proliferation and metastasis,and induces apoptosis and autophagy.These findings indicate that CRT0066101 is a promising small-mole-cule inhibitor for targeted HCC therapy with potential clinical applications.
