pubmed.ncbi.nlm.nih.gov

Caulis Spatholobi extracts inhibit osteosarcoma growth and metastasis through suppression of CXCR4/PI3K/AKT signaling - PubMed

  • ️Sun Jan 01 2023

Caulis Spatholobi extracts inhibit osteosarcoma growth and metastasis through suppression of CXCR4/PI3K/AKT signaling

Yang Jiang et al. J Orthop Surg Res. 2023.

Abstract

Background: The therapeutic potential of Caulis Spatholobi (CS) extracts against various cancers has been well documented, yet its impact and mechanism in osteosarcoma (OS) remain unexplored. This study aims to elucidate the effects of CS extracts on the growth and metastasis of OS, along with its underlying molecular mechanism.

Methods: The impact of CS extracts on the proliferative potential of two OS cell lines (Saos-2 and U2OS) was assessed using MTT and colony-formation assays. Additionally, the migratory and invasive capacities of OS cells were investigated through Transwell assays. The modulation of CXCR4 expression by CS extracts was evaluated using qRT-PCR and Western blotting. Furthermore, the influence of CS extracts on the activation of PI3K/Akt signaling was determined through Western blotting.

Results: CS extracts exhibited a dose- and time-dependent inhibition of proliferation and colony formation in OS cells. Notably, CXCR4 expression was prominently observed in Saos-2 and U2OS, and treatment with CS extracts led to a dose-dependently reduction in CXCR4 levels. Silencing CXCR4 or inhibiting its function diminished the migratory and invasive capacities of OS cells. Conversely, the CS extracts induced suppression of OS cell migration and invasion was counteracted by CXCR4 overexpression. Mechanistically, CS extracts repressed PI3K/AKT signaling in OS cells by downregulating CXCR4 expression.

Conclusions: CS extracts mitigate the CXCR4/PI3K/AKT signaling-mediated growth and metastasis capacities of OS cells, thus might play an anti-tumor role in OS.

Keywords: CXCR4; Caulis Spatholobi; Metastasis; Osteosarcoma; PI3K/AKT axis.

© 2023. The Author(s).

PubMed Disclaimer

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1

CS inhibits OS cell proliferation. A Suppression rates of Saos-2 were ascertained via MTT assay subsequent to exposure to 50, 100, 200, 400, and 800 µg/mL CS for 24, 48, and 72 h. B The half-maximum inhibitory concentration (IC50) was gauged at 24, 48, and 72 h. In comparison with the 24-h group, aP < 0.05; compared with 48 h group, bP < 0.05. C Saos-2 and U2OS cell proliferation following treatment with 0, 50, 100, and 200 µg/mL CS was detected by colony formation assay. Compared with 0 µg/mL group, aP < 0.05; Compared with 50 µg/mL group, bP < 0.05; compared with 100 µg/mL group, cP < 0.05. N = 3; data are presented as the mean ± standard deviation. CS: Caulis Spatholobi

Fig. 2
Fig. 2

CS diminishes CXCR4 expression in OS cells. The mRNA and protein expression of CXCR4 in hFOB1.19, Saos-2 and U2OS were assessed using RT-qPCR (A) and Western blotting (B), respectively. Compared with hFOB1.19 group, aP < 0.05. (C) RT-qPCR quantified CXCR4 expression in Saos-2 and U2OS treated with 50, 100 and 200 µg/mL CS for 24 h. Compared with 0 µg/mL group, aP < 0.05; Compared with 50 µg/mL group, bP < 0.05; compared with 100 µg/mL group, cP < 0.05. N = 3; the results are described as the mean ± SD. CS: Caulis Spatholobi

Fig. 3
Fig. 3

CXCR4 knockout suppresses OS cell migration and invasiveness. RT-qPCR A and B Western blotting determined CXCR4 expression in Saos-2 and U2OS treated with AMD3100 or transfected with si-CXCR4. (C) MTT assay detected Saos-2 and U2OS cell viability after AMD3100 treatment or si-CXCR4 transfection. D–E Transwell analysis was performed to observe the migration and invasiveness of Saos-2 and U2OS treated with AMD3100 or transfected with si-CXCR4. N = 3; the results are represented by the mean ± standard deviation. In comparison with the control group or si-NC group, aP < 0.05. CS, Caulis Spatholobi

Fig. 4
Fig. 4

CS attenuates OS cell migration and invasiveness by downregulating CXCR4. Saos-2 and U2OS were transfected with oe-CXCR4, and then treated with 200 μg/mL CS for 24 h. RT-qPCR (A) and Western blotting (B) quantified CXCR4 mRNA and protein expression, respectively. C, D Transwell assay tested cell migration and invasiveness. N = 3; the results are expressed as the mean ± standard deviation. Compared with oe-NC group, aP < 0.05; compared with oe-NC + CS group, bP < 0.05; compared with oe-CXCR4 + CS group, cP < 0.05. CS: Caulis Spatholobi

Fig. 5
Fig. 5

CS deactivates PI3K/AKT axis by down-regulating CXCR4. A, B Western blotting analysis was employed to measure p-PI3K, and p-AKT protein levels in Saos-2 and U2OS induced by CS (200 μg/mL, 24 h) or/and transfected by oe-CXCR4. N = 3; the data are calculated as the mean ± standard deviation. Compared with oe-NC group, aP < 0.05; compared with oe-NC + CS group, bP < 0.05; compared with oe-CXCR4 + CS group, cP < 0.05. CS: Caulis Spatholobi; p: phosphorylated

Similar articles

References

    1. Tang L, Hu H, Zhou Y, Huang Y, Wang Y, Zhang Y, et al. Expression and clinical significance of ACTA2 in osteosarcoma tissue. Oncologie. 2022;24(4):913–925. doi: 10.32604/oncologie.2022.026296. - DOI
    1. Corre I, Verrecchia F, Crenn V, Redini F, Trichet V. The osteosarcoma microenvironment: a complex but targetable ecosystem. Cells. 2020;9(4):976. doi: 10.3390/cells9040976. - DOI - PMC - PubMed
    1. Jafari F, Javdansirat S, Sanaie S, Naseri A, Shamekh A, Rostamzadeh D, et al. Osteosarcoma: a comprehensive review of management and treatment strategies. Ann Diagn Pathol. 2020;49:151654. doi: 10.1016/j.anndiagpath.2020.151654. - DOI - PubMed
    1. Smeland S, Bielack SS, Whelan J, Bernstein M, Hogendoorn P, Krailo MD, et al. Survival and prognosis with osteosarcoma: outcomes in more than 2000 patients in the EURAMOS-1 (European and American Osteosarcoma Study) cohort. Eur J Cancer (Oxford, England: 1990) 2019;109:36–50. doi: 10.1016/j.ejca.2018.11.027. - DOI - PMC - PubMed
    1. Anderson ME. Update on survival in osteosarcoma. Orthop Clin North Am. 2016;47(1):283–292. doi: 10.1016/j.ocl.2015.08.022. - DOI - PubMed

MeSH terms

Substances