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Rhodiola Crenulata Inhibits the Wnt/ß-catenin Signaling in the Treatment of Glioblastoma Multiforme
Maria Carmen Mora, MD2,3; Kaitlyn Wong, MD, MPH 2,3; Michael Tirabassi,MD1; Richard Arenas, MD 2,3,4; Sallie Schneider, PhD3,4
1Baystate Children’s Hospital, Tufts University School of Medicine, Springfield, MA; 2Baystate Medical Center, Tufts University School of Medicine, Springfield, MA; 3Pioneer Valley Life Sciences Institute, Springfield, MA; 4University of Massachusetts Amherst, Amherst, MA

Background: The purpose of this study is to determine if Rhodiola crenulata extract exhibits anti-neoplastic properties on Glioblastoma Multiforme (GBM) in-vitro.

Methods: Human U-87MG GBM line was pretreated with 200ug/ml of RC or vehicle control for 24, 48, 72, and 96 hours. Cell proliferation was then measured using an MTS assay. Neurosphere formation was evaluated by phase contrast microscopy comparing treatment with 100ug/ml RC to vehicle control. Expressions of ß-catenin and Glial fibrillary acidic protein (GFAP), a protein marker of differentiation, were measured with immunocytochemistry after 72 hours of pretreatment with 200ug/ml RC. To assess transcriptional activity of the Wnt/ß-Catenin axis, a luciferase-ß-Catenin-reporter assay was conducted. After transfection, cells were treated with 200ug/ml of RC or vehicle control and a dual luciferase-ß-Catenin-reporter assay was performed to quantify luciferase activity.

Results: MTS assay revealed a decrease in proliferation with RC therapy starting at 48 hours (p=0.01) and further reduction observed at 96 hours (p=0.0006). Neurosphere formation was eliminated after treatment with RC (Figure 1A-B). Immunocytochemistry revealed that RC induced GFAP expression (Figure1C-D) and decreased nuclear expression of ß-catenin. Luciferase assay performed on luciferase-ß-Catenin-reporter transfected cells revealed decreased Wnt promoter activity (p=0.0041) following treatment with RC after 72 hours.

Conclusion: Rhodiola crenulata extract effectively suppresses proliferation, stimulates differentiation, and eliminates tumorsphere formation of GBM cells in-vitro. The effects observed are likely secondary to the inhibition of the Wnt/ß-catenin signaling pathway.

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