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[Targeted inhibition of WISP1 enhanced radiosensitivity in glioma cells]
Objective:
To explore the effects of differencial expression of WISP1 upon the radiosenstivity and migration in glioma cell lines M059K and M059J, as well as WISP1 knockdown cell line M059K-WISP1-.
Methods:
The expression of WISP1 in different cell lines was detected by Western blot assay. The lentivirus mediated-RNA interference technology was employed to knockdown the endogenous expression of gene WISP1 in human glioma cells(M059K). A stable cell line was established by infecting M059K cells with the lentivirus particles, the transfection efficiency was examined by fluorescence microscopy. The radiosensitivity and migration ability of cells were assessed by Colony-forming and Scratch Wound Assays respectively.
Results:
WISP1 was highly expressed in M059K cells compared with M059J cells. M059K cells were less radiosensitive than M059J cells , and had a better ability of migration. M059K cells were transfected into by the lentiviral vector pGLV3/H1/GFP+puro vector -WISP1. And M059K-WISP1- cell line with stable WISP1 downregulation was established successfully. Western blot demonstrated significantly downregulated WISP1 expression in M059K-WISP1- cells, which showed obviously improved radiosensitivity in Colony-forming assay and suppressed migration ability in Scratch Wound Assay. Colony-forming assay based on the muti-target/single-hit model indicated that radiosensitivity of M059K-WISP1- was significantly decreased.
Conclusion:
Compared with WISP1 low expression cell line M059J , WISP1 high expression cell line M059K was less radiosensitive and had a better ability of migration.We have successfully established a glioma cell line M059K-WISP1- stably downregulating WISP1, which shows a better radiosensitivity and a lowered migration activity in vitro. This may provide a novel therapeutic target for the treatment of human glioma.
Insights
High WISP1 expression in glioma cells (M059K) correlated with reduced radiosensitivity and increased migration. Downregulating WISP1 in M059K cells (M059K-WISP1-) improved radiosensitivity and decreased migration, suggesting WISP1 as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Radiotherapy Research
Background:
- Glioma is a primary brain tumor with variable radiosensitivity.
- WISP1 (Wnt1-inducible signaling pathway protein 1) role in cancer progression is increasingly recognized.
- Differential expression of WISP1 may influence glioma cell behavior.
Purpose of the Study:
- To investigate the impact of WISP1 expression levels on radiosensitivity and migration in M059K and M059J glioma cell lines.
- To establish and characterize a M059K cell line with stable WISP1 knockdown (M059K-WISP1-).
Main Methods:
- Western blot analysis to determine WISP1 expression in M059K and M059J cells.
- Lentivirus-mediated RNA interference to create a stable WISP1-knockdown M059K cell line (M059K-WISP1-).
- Colony-forming and Scratch Wound Assays to evaluate radiosensitivity and cell migration, respectively.
Main Results:
- M059K cells exhibited higher WISP1 expression, lower radiosensitivity, and greater migration compared to M059J cells.
- Stable WISP1 downregulation was confirmed in the M059K-WISP1- cell line via Western blot.
- The M059K-WISP1- cell line demonstrated significantly improved radiosensitivity and suppressed migration ability in vitro.
Conclusions:
- High WISP1 expression is associated with radioresistance and enhanced migration in M059K glioma cells.
- Stable WISP1 knockdown in M059K cells (M059K-WISP1-) significantly increases radiosensitivity and reduces migration.
- WISP1 represents a potential therapeutic target for enhancing glioma treatment efficacy.
