DataSheet1_Resibufogenin Targets the ATP1A1 Signaling Cascade to Induce G2/M Phase Arrest and Inhibit Invasion in Glioma.pdf

Resibufogenin (RB) is a major active ingredient in the traditional Chinese medicine Chansu and has garnered considerable attention for its efficacy in the treatment of cancer. However, the anticancer effects and underlying mechanisms of RB on glioblastoma (GBM) remain unknown. Here, we found that RB induced G2/M phase arrest and inhibited invasion in a primary GBM cell line, P3#GBM, and two GBM cell lines, U251 and A172. Subsequently, we demonstrated that RB-induced G2/M phase arrest occurred through downregulation of CDC25C and upregulation of p21, which was caused by activation of the MAPK/ERK pathway, and that RB inhibited GBM invasion by elevating intercellular Ca2+ to suppress the Src/FAK/Paxillin focal adhesion pathway. Intriguingly, we confirmed that upon RB binding to ATP1A1, Na+-K+-ATPase was activated as a receptor and then triggered the intracellular MAPK/ERK pathway and Ca2+-mediated Src/FAK/Paxillin focal adhesion pathway, which led to G2/M phase arrest and inhibited the invasion of GBM cells. Taken together, our findings reveal the antitumor mechanism of RB by targeting the ATP1A1 signaling cascade and two key signaling pathways and highlight the potential of RB as a new class of promising anticancer agents.</p

Additional file 1 of Valtrate, an iridoid compound in Valeriana, elicits anti-glioblastoma activity through inhibition of the PDGFRA/MEK/ERK signaling pathway

Additional file 1: Figure S1. Valtrate inhibits cell proliferation in GBM cells. Figure S2. Valtrate promotes apoptosis in GBM cells via the mitochondrial pathway. Figure S3. Valtrate suppresses migration and invasion of GBM cells. Figure S4. PDGFRA is a potential target downregulated by valtrate in GBM cells. (A) Volcano plot showing the up- and downregulated genes, red and blue colors, respectively, obtained from RNA-seq analysis. Cells were treated with valtrate (U251: 2 μM, GBM#P3: 0.5 μM) for 48 h and RNA was isolated and sequenced. (B) Cell viability of LN229-PDGFRA-OE under the conditions indicated as determined with the CCK-8 assay. (C) Representative images of EdU assays for U251- and LN229-PDGFRA-OE cells under the conditions indicated. Scale bar, 50 μm. (D) Flow cytometry to detect the percentage of apoptotic U251- and GBM#P3-PDGFRA-OE cells under the conditions indicated as determined with annexin V-FITC and PI staining. (E) Representative images of 3D invasion assay for U251- and GBM#P3-PDGFRA-OE PDGFRA cells under the conditions indicated, with or without valtrate. Scale bar, 200 μm. All data are expressed as the mean ± SD of values from triplicate experiments and the differences between groups were analyzed with the Student’s t-test. *p < 0.05. Figure S5. Valtrate elicits anti-GBM activity through inhibition of the PDGFRA/MEK/ERK signaling pathway. Figure S6. Valtrate exerts its antitumor effects in vivo

Knockdown of NUSAP1 inhibits cell proliferation and invasion through downregulation of TOP2A in human glioblastoma

Nucleolar and spindle associated protein 1 (NUSAP1), an indispensable mitotic regulator, has been reported to be involved in the development, progression, and metastasis of several types of cancer. Here, we investigated the expression and biological function of NUSAP1 in human glioblastoma (GBM), an aggressive brain tumor type with largely ineffective treatment options. Analysis of the molecular data in CGGA, TCGA and Rembrandt datasets demonstrated that NUSAP1 was significantly upregulated in GBM relative to low grade gliomas and non-neoplastic brain tissue samples. Kaplan-Meier analysis indicated that patients with tumors showing high NUSAP1 expression exhibited significantly poorer survival in both CGGA (P = 0.002) and Rembrandt cohorts (P = 0.017). Analysis of RNA sequencing data from P3-cells with stable knockdown of NUSAP1 revealed topoisomerase 2A (TOP2A) as a possible molecule downregulated by the loss of NUSAP1. Molecular analysis of the CGGA data revealed a strong correlation between NUSAP1 and TOP2A expression in primary gliomas and recurrent gliomas samples. SiRNA knockdown of either NUSAP1 or TOP2A in U251, T98 and GBM derived patient P3 cells inhibited GBM cell proliferation and invasion, and induced cell apoptosis. Finally, stable knockdown of NUSAP1 with shRNA led to decreased tumor growth in an orthotopic xenograft model of GBM in mice. Taken together, NUSAP1 gene silencing induced apoptosis possibly through the downregulation of the candidate downstream molecule TOP2A. Interference with the expression of NUSAP1 might therefore inhibit malignant progression in GBM, and NUSAP1 might thus serve as a promising molecular target for GBM treatment.</p

Additional file 1 of The dopamine receptor D1 inhibitor, SKF83566, suppresses GBM stemness and invasion through the DRD1-c-Myc-UHRF1 interactions

Additional file 1: 21-day mature rat brain organoids were co-cultured with GFP-labeled tumor GSC spheres in round well low-attachment 96-well plates for 24 h, 48 h or 72 h. To isolate invading GFP-labeled tumor cells from the main tumor mass, the co-cultures were cut in half under a dissecting microscope