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. 2024 Jan 7;25(2):759.
doi: 10.3390/ijms25020759.

Furanocoumarins as Enhancers of Antitumor Potential of Sorafenib and LY294002 toward Human Glioma Cells In Vitro

Joanna Sumorek-Wiadro  1 , Adrian Zając  1 , Krystyna Skalicka-Woźniak  2 , Wojciech Rzeski  1   3 , Joanna Jakubowicz-Gil  1
Affiliations

Furanocoumarins as Enhancers of Antitumor Potential of Sorafenib and LY294002 toward Human Glioma Cells In Vitro

Joanna Sumorek-Wiadro et al. Int J Mol Sci. .

Abstract

Furanocoumarins are naturally occurring compounds in the plant world, characterized by low molecular weight, simple chemical structure, and high solubility in most organic solvents. Additionally, they have a broad spectrum of activity, and their properties depend on the location and type of attached substituents. Therefore, the aim of our study was to investigate the anticancer activity of furanocoumarins (imperatorin, isoimperatorin, bergapten, and xanthotoxin) in relation to human glioblastoma multiforme (T98G) and anaplastic astrocytoma (MOGGCCM) cell lines. The tested compounds were used for the first time in combination with LY294002 (PI3K inhibitor) and sorafenib (Raf inhibitor). Apoptosis, autophagy, and necrosis were identified microscopically after straining with Hoechst 33342, acridine orange, and propidium iodide, respectively. The levels of caspase 3 and Beclin 1 were estimated by immunoblotting and for the blocking of Raf and PI3K kinases, the transfection with specific siRNA was used. The scratch test was used to assess the migration potential of glioma cells. Our studies showed that the anticancer activity of furanocoumarins strictly depended on the presence, type, and location of substituents. The obtained results suggest that achieving higher pro-apoptotic activity is determined by the presence of an isoprenyl moiety at the C8 position of the coumarin skeleton. In both anaplastic astrocytoma and glioblastoma, imperatorin was the most effective in induction apoptosis. Furthermore, the usage of imperatorin, alone and in combination with sorafenib or LY294002, decreased the migratory potential of MOGGCCM and T98G cells.

Keywords: LY294002; apoptosis; bergapten; furanocoumarins; gliomas; imperatorin; isoimperatorin; programmed cell death; sorafenib; xanthotoxin.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Structural formulas of bergapten (a), xanthotoxin (b), isoimperatorin (c), and imperatorin (d).
Figure 2
Figure 2
Type and level of cell death observed in MOGGCCM cells after 24 h incubation with different concentrations of bergapten (a), xanthotoxin (b), imperatorin (c), and isoimperatorin (d). * p < 0.05 compared to control.
Figure 3
Figure 3
Type and level of cell death observed in T98G cells after 24 h incubation with different concentrations of bergapten (a), xanthotoxin (b), imperatorin (c), and isoimperatorin (d). * p < 0.05 compared to control.
Figure 4
Figure 4
Representative photos of cells stained with T98G (a,b,e,f), OLN-93 (c,d), Hoechst 33342 and propidium iodide (a,d), and orange acridine (e,f). Apoptosis (b) with characteristic apoptotic bodies (white arrows) and autophagy (f) with AVOs (white arrows). C—control; I—imperatorin; LY—LY294002.
Figure 5
Figure 5
Evaluation of level of cell death observed in MOGGCCM (a), T98G (b), and OLN-93 (c) cells after 24 h incubation with bergapten (B), xanthotoxin (X), imperatorin (I), isoimperatorin (IS), and sorafenib (S) in single or in combination. * p < 0.05 compared to control.
Figure 6
Figure 6
Evaluation of level of cell death observed in MOGGCCM (a), T98G (b), and OLN-93 (c) cells after 24 h incubation with bergapten (B), xanthotoxin (X), imperatorin (I), isoimperatorin (IS), and LY294002 (LY) in single or in combination. * p < 0.05 compared to control.
Figure 7
Figure 7
Evaluation of procaspase 3 (a,b), Beclin 1 (c,d) level, and activity of caspase 3 in MOGGCCM (a,c,e), T98G (b,d,f) cells after 24 h incubation with imperatorin (I), sorafenib (S), and LY294002 (LY) in single or in combination. * p < 0.05 compared to control; whole Western blot membranes attached as Supplementary Information.
Figure 8
Figure 8
Level of cell death in MOGGCCM and T98G cells transfected with siPI3K (a,b), and siRAF-1 (c,d) after 24 h incubation with imperatorin (I), LY294002 (LY), and/or sorafenib (S) in single or in combination. C—control cells; TR—transfection reagent; * p < 0.05 compared to control; photos of transfection efficiency attached as Supplementary Information.
Figure 9
Figure 9
Migration potential of the MOGGCCM cells upon imperatorin (I) and sorafenib (S) or LY294002 (LY) presented as the percent of cells within the woud (WA; red lines on the figures). C-control; * p < 0.05.
Figure 10
Figure 10
Migration potential of the T98G cells upon imperatorin (I) and sorafenib (S) or LY294002 (LY) presented as the percent of cells within the woud (WA; red lines on the figures). C-control; * p < 0.05.
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References

    1. Brito C., Azevedo A., Esteves S., Marques A.R., Martins C., Costa I., Mafra M., Bravo Marques J.M., Roque L., Pojo M. Clinical insights gained by refining the 2016 WHO classification of diffuse gliomas with: EGFR amplification, TERT mutations, PTEN deletion and MGMT methylation. BMC Cancer. 2019;19:1–14. doi: 10.1186/s12885-019-6177-0. - DOI - PMC - PubMed
    1. Szala S., Jarosz M., Smolarczyk R., Cichoń T. Błędne koła glejaków: Unaczynienie i inwazyjność. Post. Hig. 2012;66:888–900. doi: 10.5604/17322693.1019657. - DOI - PubMed
    1. Nakada M., Kita D., Watanabe T., Hayashi Y., Teng L., Pyko I.V., Hamada J. Aberrant signaling pathways in glioma. Cancers. 2011;3:3242–3278. doi: 10.3390/cancers3033242. - DOI - PMC - PubMed
    1. Annunziata F., Pinna C., Dallavalle S., Tamborini L., Pinto A. An overview of coumarin as a versatile and readily accessible scaffold with broad-ranging biological activities. Int. J. Mol. Sci. 2020;21:4618. doi: 10.3390/ijms21134618. - DOI - PMC - PubMed
    1. Wang G., Sun S., Wu B., Liu J. Coumarins as Potential Anti-drug Resistant Cancer Agents: A Mini Review. Top. Med. Chem. 2021;21:1725–1736. doi: 10.2174/1568026620999201113110041. - DOI - PubMed