Moschamine inhibits proliferation of glioblastoma cells via cell cycle arrest and apoptosis

Abstract

Glioblastoma is the most common and most malignant primary brain tumor with a median survival of 15 months. Moschamine is an indole alkaloid that has a serotoninergic and cyclooxygenase inhibitory effect. In this study, we sought to determine whether moschamine could exert cytotoxic and cytostatic effects on glioma cells in vitro. Moschamine was tested for toxicity in zebrafish. We investigated the effect of moschamine on U251MG and T98G glioblastoma cell lines. Viability and proliferation of the cells were examined with trypan blue exclusion assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the xCELLigence system. Apoptosis (annexin-propidium iodide), cell cycle, and CD24/CD44/CD56/CD15 expression were tested with flow cytometry. Treatment with moschamine significantly reduced cell viability in both cell lines tested. Induction of cell death and cell cycle arrest was confirmed with flow cytometry in both cell lines. After treatment with moschamine, there was a dose-dependent decrease in CD24 and CD44 expression, whereas there was no change in CD56 and CD15 expression in T98G cell line. The zebrafish mortality on the fifth post-fertilization day was zero even for 1 mM of moschamine concentration. The treatment of glioblastoma cell lines with moschamine may represent a novel strategy for targeting glioblastoma.

The Past, Present and Future of Flow Cytometry in Central Nervous System Malignancies

“…Our research group has been working towards novel therapeutics of GBM, which are based on natural products. Specifically, on the basis of experimental results that include FC methodologies, we have found that difluoromethylornithine (DFMO) [ 96 ] as well as the natural substances moschamine [ 97 ], n-p-coumaroyl-serotonin [ 98 ], and deglucohellebrin [ 99 ] exhibit significant antiglioma activity in vitro and low cytotoxity in vivo, as shown in a zebrafish embryo model.…”

“…DNA content analysis revealed several mechanisms of action for different antiglioma agents. For example, N-(p-coumaroyl)-serotonin resulted in both S and G2/M phase arrest, moschamine resulted mostly in S phase arrest and deglucohellebrin in G2/M phase arrest [ 97 , 98 , 99 ]. Several other studies have used in DNA content analysis quantification in several CNSM treatments, revealing, among others, the antiglioma effects of curcumin [ 100 ], quercetin [ 101 ], glycyrrhizic acid [ 102 ], and palbociclib [ 103 ].…”

Patient-Derived Xenotransplant of CNS Neoplasms in Zebrafish: A Systematic Review

“…Most of the biological characteristics such as proliferation, apoptosis, and drug screening, among others, have been evaluated in vitro using cell lines [ 70 , 107 , 108 , 109 , 110 , 111 , 112 ]. Although they have been instrumental in understanding tumor behavior, the mere fact of being a cell line does not represent the actual heterogeneity of a tumor sample from a patient.…”

Haloperidol Induced Cell Cycle Arrest and Apoptosis in Glioblastoma Cells

“…The activity of caspase-8 was quantified with the Fluorescein Active Caspase-8 Staining Kit (Abnova, Taiwan) as described previously [ 15 ]. For the assessment of the cluster of differentiation (CD) expression, specifically CD24/CD44, we used the FITC mouse anti-human CD24 (ML5), PE mouse anti-human CD24 (ML5), and FITC mouse anti-human CD44 (Leu-44) (all from BD Pharmingen) as described previously in detail [ 16 ]. Approximately 10,000 cells were seeded in 24-well plates and after 24 h were exposed to a concentration of 100 μΜ of haloperidol, for another 72 h. The cells were then dissociated by trypsinization, washed twice with PBS and in order to block Fc receptors, incubated with 10% human serum for 20 min on ice.…”