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. 2022 May-Jun;36(3):1136-1143.
doi: 10.21873/invivo.12812.

Reinforcement of Sorafenib Anti-osteosarcoma Effect by Amentoflavone Is Associated With the Induction of Apoptosis and Inactivation of ERK/NF-κB

Chun-Min Su #  1 , Chi-Huan Li #  2 , Meng-Chu Huang #  3 , Po-Fu Yueh  4   5 , Fei-Ting Hsu  4 , Rong-Fong Lin  6 , Li-Cho Hsu  7
Affiliations

Reinforcement of Sorafenib Anti-osteosarcoma Effect by Amentoflavone Is Associated With the Induction of Apoptosis and Inactivation of ERK/NF-κB

Chun-Min Su et al. In Vivo. 2022 May-Jun.

Abstract

Background/aim: Sorafenib has been reported to show anti-osteosarcoma (anti-OS) efficacy by inhibiting metastasis; however, a phase II trial suggested that further combination with other agents could be necessary to achieve permanent remission. Herein, we aimed to identify whether amentoflavone, an abundant natural bioflavonoid found in many medicinal plants, can improve the treatment efficacy of sorafenib in OS.

Materials and methods: Cell viability, metastasis, apoptosis, and nuclear translocation of NF-κB after amentoflavone combined with sorafenib were assayed by MTT, transwell migration/invasion, western blotting, flow cytometry, and immunofluorescence staining, respectively.

Results: The sorafenib-induced cytotoxicity and apoptosis of U-2 OS was enhanced by combining treatment with QNZ (NF-κB inhibitor) or amentoflavone. NF-κB nuclear translocation, NF-κB phosphorylation, and metastasis capacity of U-2 OS cells were inhibited by amentoflavone combined with sorafenib.

Conclusion: Amentoflavone may sensitize OS to sorafenib treatment by inducing intrinsic and extrinsic apoptosis and inhibiting ERK/NF-κB signaling transduction.

Keywords: Amentoflavone; ERK; NF-κB; apoptosis.

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Conflict of interest statement

The Authors declare no competing financial interests regarding this study.

Figures

Figure 1
Figure 1. The induction of sorafenib-induced toxicity and sorafenib-suppressed migration and invasion by amentoflavone in U-2 OS cells. U-2 OS cells were treated with (A) sorafenib (0-30 μM), (B) amentoflavone (0-200 μM) or (C) the combination of both for 24 h, and then assayed by MTT. (D) The CI index of the combination treatment is displayed. The (E) stained transwell images of migration and invasion and (F) the quantification of data are presented. (G) The western blotting results of MMP-9 and VEGF after treatment are presented. (*p<0.05, **p<0.01 vs. 0 μM of sorafenib or 0 μM of amentoflavone; $p<0.05, $$p<0.01 vs. 0 μM of amentoflavone).
Figure 2
Figure 2. Both the extrinsic and intrinsic apoptosis signaling pathways induced by sorafenib were reinforced by amentoflavone. U-2 OS cells were treated with 100 μM of amentoflavone, 20 μM of sorafenib, and the combination of both for 24 h and assayed by using flow cytometry. The (A) activation pattern of cleaved caspase-3 and cleaved caspase-8 and (B-C) quantitation analysis of their activation are displayed. The (D) activation pattern of Fas and Fas-L and (E-F) quantitation analysis of their activation are presented. The (G) activation pattern of cleaved caspase-9 and the loss of ΔΨm and (H-I) quantitation analysis of their activation are displayed. (J) The western blotting results of XIAP and C-FLIP after treatment are presented. *p<0.05, **p<0.01, ***p<0.005 vs. CTRL; $p<0.05, $$p<0.01 vs. 100 μM of amentoflavone and 20 μM of sorafenib.
Figure 3
Figure 3. Suppression of ERK/NF-ĸB by amentoflavone may reinforce the sorafenib-induced toxicity of U-2 OS cells. U-2 OS cells were treated with (A) QNZ (0-4 μM) or (B) the combination of 0.25 QNZ μM with sorafenib (0-30 μM) for 24 h and then assayed by MTT. (C) The CI value of QNZ combined with sorafenib is displayed. The nuclear translocation (D) shown by immunofluorescence staining imaging and (E) quantification analysis are presented. (J) The western blotting results of ERK (Thr202/Tyr204), ERK, NF-ĸB (Ser536), NF-ĸB, XIAP, and C-FLIP after treatment are presented.
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References

    1. Chang PY, Hsieh MJ, Hsieh YS, Chen PN, Yang JS, Lo FC, Yang SF, Lu KH. Tricetin inhibits human osteosarcoma cells metastasis by transcriptionally repressing MMP-9 via p38 and Akt pathways. Environ Toxicol. 2017;32(8):2032–2040. doi: 10.1002/tox.22380. - DOI - PubMed
    1. Pan PJ, Tsai JJ, Liu YC. Amentoflavone inhibits metastatic potential through suppression of ERK/NF-ĸB activation in osteosarcoma U2OS cells. Anticancer Res. 2017;37(9):4911–4918. doi: 10.21873/anticanres.11900. - DOI - PubMed
    1. Czarnecka AM, Synoradzki K, Firlej W, Bartnik E, Sobczuk P, Fiedorowicz M, Grieb P, Rutkowski P. Molecular biology of osteosarcoma. Cancers (Basel) 2020;12(8):2130. doi: 10.3390/cancers12082130. - DOI - PMC - PubMed
    1. Prudowsky ZD, Yustein JT. Recent insights into therapy resistance in osteosarcoma. Cancers (Basel) 2020;13(1):83. doi: 10.3390/cancers13010083. - DOI - PMC - PubMed
    1. Tian Z, Niu X, Yao W. Receptor tyrosine kinases in osteosarcoma treatment: which is the key target. Front Oncol. 2020;10:1642. doi: 10.3389/fonc.2020.01642. - DOI - PMC - PubMed