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. 2019 Apr;7(1):16-24.
doi: 10.14791/btrt.2019.7.e28.

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

Ki Su Park #  1 , Sang Youl Yoon #  1 , Seong Hyun Park  1 , Jeong Hyun Hwang  2
Affiliations

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

Ki Su Park et al. Brain Tumor Res Treat. 2019 Apr.

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.

Keywords: Curcumin; Fascin; Glioblastoma; STAT3 transcription factor.

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Conflict of interest statement

The authors have no potential conflicts of interest.

Figures

Fig. 1
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
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
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
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
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
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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