Brain Tumor Res Treat.  2019 Apr;7(1):16-24. 10.14791/btrt.2019.7.e28.

Anti-Migration and Anti-Invasion Effects of Curcumin via Suppression of Fascin Expression in Glioblastoma Cells

• Park KS
• Yoon SY
• Park SH
• Hwang JH

• KMID: 2444791
• DOI: http://doi.org/10.14791/btrt.2019.7.e28

Abstract

BACKGROUND
The natural compound curcumin was known to inhibit migration and invasion of glioblastoma (GBM) cells. Fascin, a kind of actin-binding proteins, is correlated with migration and invasion of GBM cells. The purpose of this study was to investigate anti-migration and anti-invasion effects of curcumin via suppression of fascin expression in GBM cells.
METHODS
U87 cell line was used as an experimental model of GBM. Fascin was quantified by Western blot analysis. And, the signal transducer and activator of transcription 3 (STAT3), known to play an important role in migration and invasion of tumor cells, were analyzed by sandwich-ELISA. Migration and invasion capacities were assessed by attachment, migration and invasion assays. Cellular morphology was demonstrated by immunofluorescence.
RESULTS
At various concentrations of curcumin and exposure times, fascin expression decreased. After temporarily exposure to 10 µM/L curcumin during 6 hours as less invasive concentration and time, fascin expression temporarily decreased at 12 hours (18.4%, p=0.024), and since then recovered. And, the change of phosphrylated STAT3 level also reflected the temporarily decreased pattern of fascin expression at 12 hours (19.7%, p=0.010). Attachment, migration, and invasion capacities consistently decreased at 6, 12, and 24 hours. And, immunofluorescence showed the change of shape and the reduction of filopodia formation in cells.
CONCLUSION
Curcumin is likely to suppress the fascin expression in GBM cells, and this might be a possible mechanism for anti-migration and anti-invasion effects of Curcumin via inhibition of STAT3 phosphorylation.

Keyword

Glioblastoma; Fascin; Curcumin; STAT3 transcription factor

MeSH Terms

Blotting, Western
Cell Line
Curcumin*
Emigration and Immigration
Fluorescent Antibody Technique
Glioblastoma*
Microfilament Proteins
Models, Theoretical
Phosphorylation
Pseudopodia
STAT3 Transcription Factor
Curcumin
Microfilament Proteins
STAT3 Transcription Factor

Figure

• 

  Fig. 1 MTS assay for cell viability. A: At 10 µM/L and 20 µM/L concentrations of curcumin, cell viabilities showed minimal decreases (10 µM/L: 80.8%, 20 µM/L: 78.2%). B: The cell viabilities at 10 µM/L and 20 µM/L curcumin for 6 hours were 93.9% and 90.5%, respectively. *p<0.05, **p<0.001 compared to the control group.

• 

  Fig. 2 Western blot analysis for quantification of fascin expression and sandwich ELISA for quantification of STAT3. A: Fascin expression temporarily decreased at 12 hours (18.4%, p=0.024), but gradually recovered after 12 hours. B: The change of phosphorylated STAT3 level also reflected temporarily decreased patterns of fascin expression at 12 hours (19.7%, p=0.010). Whereas concentrations of total STAT3 according to a time sequence were similar. Data were presented as the mean±SD. *p<0.05 compared to the control group. STAT3, signal transducer and activator of transcription 3.

• 

  Fig. 3 Attachment assay. The attachment ability of glioblastoma cells showed a statistically significant decrease after 6 hours (20 µM/L curcumin at 12 hours: 14.7%, p=0.001; 10 and 20 µM/L curcumin at 24 hours: 12.6% and 24.4%, p=0.001 and p<0.001). Data were presented as the mean±SD. *p<0.05, **p<0.001 compared to the control group.

• 

  Fig. 4 Migration assay and microscopic images (magnification ×100). A: The migration ability of GBM cells showed a statistically significant decline after 6 hours, and the closure percentage of GBM cells showed statistical differences when compared to control (10 µM/L and 20 µM/L at 12 hours: 24.2% and 58.1%, p=0.031 and p<0.001; at 10 µM/L and 20 µM/L at 24 hours: 25.0% and 54.3%, p=0.002 and p<0.001). B: After 0 µM/L curcumin during 6 hours, a photograph showed the migration capacity of GBM cells after 0, 6, 12, and 24 hours. C: After 10 µM/L curcumin for 6 hours, a photograph showed the migration capacity of GBM cells after 0, 6, 12, and 24 hours. D: After 20 µM/L curcumin for 6 hours, a photograph showed the migration capacity of GBM cells after 0, 6, 12, and 24 hours. Data were presented as the mean±SD. *p<0.05, **p<0.001 compared to the control group. GBM, glioblastoma.

• 

  Fig. 5 Invasion assay with microscopic images (magnification ×100). A: The invasion ability of GBM cells showed a statistically significant decline after 6 hours (10 µM/L and 20 µM/L at 12 hours: 28.8% and 43.2%, p=0.046 and p<0.001; 10 µM/L and 20 µM/L at 24 hours: 54.1% and 73.8%, p=0.001 and p<0.001). B: After 0 µM/L curcumin for 6 hours, a photograph showed the migration capacity of GBM cells after 6, 12, and 24 hours. C: After 10 µM/L curcumin for 6 hours, a photograph showed the migration capacity of GBM cells after 6, 12, and 24 hours. D: After 20 µM/L curcumin for 6 hours, a photograph showed the migration capacity of GBM cells after 6, 12, and 24 hours. Data were presented as the mean±SD. *p<0.05, **p<0.001 compared to the control group. GBM, glioblastoma.

• 

  Fig. 6 After temporary exposure to 0 µM/L (A) and 10 µM/L (B) curcumin for 6 hours, immunofluorescence at 6 and 24 hours showed a change in cell shape from a typical spindle form of uninfluenced cells to a more polygonal appearance, and a significant reduction in filopodia formation of glioblastoma cells (×400).

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