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PDGF-BB/PDGFRβ promotes epithelial-mesenchymal transition by affecting PI3K/AKT/mTOR-driven aerobic glycolysis in
Jia-Qi Guo1,2, Chang-Dong Wang2,3, Hu-Ying Tang1,2
1Department of Physiology, Basic Medical College, Chongqing Medical University, Chongqing, China.
Abstract:
Wilms' tumor (WT) is the most common pediatric renal malignancy. PDGFRβ belongs to the type III receptor tyrosine kinase family and is known to be involved in tumor metastasis and angiogenesis. Here, we studied the effect and underlying mechanism of PDGFRβ on WT G401 cells. Transwell assay and wound-healing assay were used to detect the effect of PDGFRβ on G401 cells invasion and migration. Western blot and immunofluorescence were used to detect the expression of EMT-related genes. The expression of PI3K/AKT/mTOR pathway proteins was detected by Western blot. The relationship between PDGFRβ and aerobic glycolysis was studied by assessing the expression of glycolysis-related enzymes detected by qRT-PCR and Western blot. The activity of HK, PK, and LDH was detected by corresponding enzyme activity kits. The concentration of lactic acid and glucose was detected by Lactic Acid Assay Kit and Glucose Assay Kit-glucose oxidase method separately. To investigate the mechanism of PDGFRβ in the development of WT, the changes of glucose and lactic acid were analyzed after blocking PI3K pathway, aerobic glycolysis, or PDGFRβ. The key enzyme was screened by Western blot and glucose metabolism experiment after HK2, PKM2, and PDK1 were inhibited. The results showed that PDGFRβ promoted the EMT process by modulating aerobic glycolysis through PI3K/AKT/mTOR pathway in which PKM2 plays a key role. Therefore, our study of the mechanism of PDGFRβ in G401 cells provides a new target for the treatment of WT.
Insights
Platelet-derived growth factor receptor beta (PDGFRβ) promotes Wilms' tumor (WT) cell invasion and migration. PDGFRβ enhances the epithelial-mesenchymal transition (EMT) process by modulating aerobic glycolysis via the PI3K/AKT/mTOR pathway, identifying PKM2 as a key enzyme.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Wilms' tumor (WT) is the most common pediatric kidney cancer.
- Platelet-derived growth factor receptor beta (PDGFRβ) is implicated in tumor metastasis and angiogenesis.
Purpose of the Study:
- To investigate the effect and mechanism of PDGFRβ in WT G401 cells.
- To explore the role of PDGFRβ in epithelial-mesenchymal transition (EMT) and aerobic glycolysis.
Main Methods:
- Transwell and wound-healing assays for cell invasion and migration.
- Western blot, immunofluorescence, and qRT-PCR for gene and protein expression.
- Enzyme activity kits and biochemical assays for glycolysis and metabolite concentrations.
Main Results:
- PDGFRβ significantly promoted G401 cell invasion and migration.
- PDGFRβ upregulated EMT-related genes and proteins.
- PDGFRβ enhanced aerobic glycolysis through the PI3K/AKT/mTOR pathway, with PKM2 identified as a key enzyme.
Conclusions:
- PDGFRβ drives WT cell progression by promoting EMT via aerobic glycolysis.
- The PI3K/AKT/mTOR pathway and PKM2 are crucial mediators of PDGFRβ's effects.
- Targeting PDGFRβ offers a potential therapeutic strategy for WT.
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