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Spheroid Assay to Measure TGF-β-induced Invasion
Published on: November 16, 2011
STC1 regulates glioblastoma migration and invasion via the TGF‑β/SMAD4 signaling pathway
1Department of Neurosurgery, Chongqing Ninth People's Hospital, Chongqing 400715, P.R. China.
Abstract:
Stanniocalcin‑1 (STC1) is involved in cancer progression; however, the function of STC1 in glioblastoma remains unknown. In the present study, the expression levels of STC1 protein in glioblastoma were detected using immunohistochemistry. The expression levels of STC1, SMAD2/3 and SMAD4 proteins, following silencing of STC1, were assessed via western blotting. EdU and Transwell assays were performed to determine the proliferation and migration ability of the cells. The mRNA expression levels of STC1, SMAD4 and microRNA (miR)‑34a were determined using quantitative PCR. The expression levels of STC1 were increased in glioblastoma tissues. STC1 revealed a significant association with poor outcome in patients with glioblastoma (P<0.05). The proliferation and invasion abilities were repressed in LN229 cells infected with LV3‑shSTC1‑1 and LV3‑shSTC1‑2 compared with LV3‑NC. By contrast, the proliferation and invasion abilities were increased in T98G cells infected with LV5‑STC1 compared with LV5‑NC (P<0.05). The expression levels of STC1, SMAD2/3 and SMAD4 were decreased in LN229 cells infected with LV3‑shSTC1‑1 and LV3‑shSTC1‑2 compared with LV3‑NC. However, the expression levels of STC1, SMAD2/3 and SMAD4 were elevated in T98G cells infected with LV5‑STC1 compared with LV5‑NC. The expression levels of miR‑34a were decreased following silencing of STC1 (P<0.05). The expression levels of SMAD4 were decreased when transfected with miR‑34a mimics (P<0.05). The luciferase activity of the wild‑type 3'untranslated region of SMAD4 was decreased following transfection with miR‑34a mimics (P<0.05). Silencing of STC1 inhibited the growth of LN229 in vivo. In conclusion, STC1 expression levels were increased in the present study, and it was revealed that STC1 regulated glioblastoma malignancy. This phenotype was observed in the SMAD2/3 and SMAD4 pathways.
Insights
Stanniocalcin-1 (STC1) is upregulated in glioblastoma and promotes cancer progression by influencing SMAD pathways. Silencing STC1 inhibits tumor growth and invasion, suggesting STC1 as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Stanniocalcin-1 (STC1) is implicated in cancer progression, but its role in glioblastoma remains unclear.
- Glioblastoma is an aggressive brain tumor with limited treatment options.
Purpose of the Study:
- To investigate the expression and function of STC1 in glioblastoma.
- To elucidate the molecular mechanisms underlying STC1's role in glioblastoma malignancy.
Main Methods:
- Immunohistochemistry to detect STC1 protein expression in glioblastoma tissues.
- Western blotting to assess protein levels of STC1, SMAD2/3, and SMAD4 after STC1 silencing.
- EdU and Transwell assays to evaluate cell proliferation and migration.
- Quantitative PCR to determine mRNA expression of STC1, SMAD4, and microRNA-34a.
- In vivo studies using LN229 xenografts to assess tumor growth inhibition.
Main Results:
- STC1 expression is significantly increased in glioblastoma tissues and associated with poor patient outcomes.
- Silencing STC1 inhibited proliferation and invasion in LN229 cells, while STC1 overexpression increased these abilities in T98G cells.
- STC1 silencing decreased SMAD2/3 and SMAD4 expression, whereas STC1 overexpression increased them.
- STC1 silencing reduced microRNA-34a levels, and microRNA-34a mimics decreased SMAD4 expression and luciferase activity, indicating a regulatory relationship.
- In vivo, STC1 silencing suppressed LN229 glioblastoma growth.
Conclusions:
- STC1 is upregulated in glioblastoma and promotes tumor malignancy.
- STC1 exerts its effects through the SMAD2/3 and SMAD4 signaling pathways.
- STC1 inhibition represents a potential therapeutic strategy for glioblastoma.
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