Proapoptotic and antimigration properties of osthole in combination with LY294002 against human glioma cells
Joanna Sumorek-Wiadro1, Justyna Kapral-Piotrowska2, Adrian Zając2
1Department of Functional Anatomy and Cytobiology, Institute of Biological Sciences, Maria Curie-Sklodowska University, Akademicka 19, 20-033, Lublin, Poland. joanna.sumorek-wiadro@mail.umcs.pl.
Abstract:
Anaplastic astrocytoma and glioblastoma multiforme are infiltrating and vascularized gliomas with a high degree of chemoresistance and metastasis. Our previous studies have shown that osthole may be of great importance in the treatment of gliomas. Therefore, in this work, for the first time, coumarin was used in combination with LY294002-an inhibitor of the PI3K-Akt/PKB-mTOR pathway, which is overly active in gliomas. MOGGCCM and T98G cells were incubated with osthole and LY294002, alone and in combination. Staining with specific fluorochromes was used to visualize cell death and the scratch test to assess the migration. The level of proteins was estimated by immunoblotting. Forming protrusions were visualized by SEM, and immunocytochemistry was used to determine the localization of proteins. Additionally, the expression of Bcl-2, beclin 1 and Raf kinase was silenced using specific siRNA. The obtained results showed that osthole in combination with LY294092 effectively inhibited the migration of glioma cells by reducing the level of metaloproteinases and Rho family proteins, as well as decreasing the level of N-cadherin. In addition, the combination of compounds induced apoptosis. New combination of compounds shows a high pro-apoptotic potential and also inhibits the migration of gliomas cells.
Insights
Osthole combined with LY294002 effectively inhibits glioma cell migration and induces apoptosis. This novel combination shows significant potential for treating aggressive brain tumors like anaplastic astrocytoma and glioblastoma multiforme.
Area of Science:
- Neuro-oncology
- Pharmacology
- Molecular Biology
Background:
- Anaplastic astrocytoma and glioblastoma multiforme are aggressive brain tumors.
- These gliomas exhibit chemoresistance and metastasis.
- The PI3K-Akt/PKB-mTOR pathway is often overactive in gliomas.
Purpose of the Study:
- To investigate the combined effect of osthole and LY294002 on glioma cells.
- To evaluate the impact on cell migration and apoptosis.
- To explore the underlying molecular mechanisms.
Main Methods:
- Glioma cells (MOGGCCM, T98G) treated with osthole and LY294002.
- Assays for cell death (staining), migration (scratch test), and protein levels (immunoblotting).
- Scanning electron microscopy (SEM) for cell morphology and immunocytochemistry for protein localization. siRNA used to silence specific protein expression.
Main Results:
- The combination of osthole and LY294002 significantly inhibited glioma cell migration.
- Reduced levels of matrix metalloproteinases and Rho family proteins were observed.
- N-cadherin levels decreased, and apoptosis was induced in glioma cells.
Conclusions:
- Osthole and LY294002 combination exhibits potent anti-migratory effects on glioma cells.
- The combination demonstrates significant pro-apoptotic potential.
- This novel therapeutic strategy warrants further investigation for glioma treatment.
