The Evaluation of Glioblastoma Cell Dissociation and Its Influence on Its Behavior

Abstract

Purpose: Primary cell lines are a valuable tool for evaluation of tumor behavior or sensitivity to anticancer treatment and appropriate dissociation of cells could preserve genomic profile of the original tissue. The main aim of our study was to compare the influence of two methods of glioblastoma multiforme (GBM) cell derivation (mechanic—MD; enzymatic—ED) on basic biological properties of thus derived cells and correlate them to the ones obtained from stabilized GBM cell line A-172. Methods: Cell proliferation and migration (xCELLigence Real-Time Cell Analysis), expression of microRNAs and protein markers (RT-PCR and Western blotting), morphology (phase contrast and fluorescent microscopy), and accumulation of temozolomide (TMZ) and its metabolite 5-aminoimidazole-4-carboxamide (AIC) inside the cells (LC-MS analysis) were carried out in five different samples of GBM (GBM1, GBM2, GBM32, GBM33, GBM34), with each of them processed by MD and ED types of isolations. The same analyses were done in the A-172 cell line too. Results: Primary GBM cells obtained by ED or MD approaches significantly differ in biological behavior and properties of these cells. Unlike in primary MD GBM cells, higher proliferation, as well as migration, was observed in primary ED GBM cells, which were also associated with the acquired mesenchymal phenotype and higher sensitivity to TMZ. Finally, the same analyses of stabilized GBM cell line A-172 revealed several important differences in measured parameters. Conclusions: GBM cells obtained by MD and ED dissociation show considerable heterogeneity, but based on our results, MD approach should be the preferred method of primary GBM cell isolation

Flubendazole exhibits anti-glioblastoma effect by inhibiting STAT3 and promoting cell cycle arrest

“…201709 S13P) and patients gave their written informed consent. Cells were isolated by mechanic dissociation as described previously 26 and all experiments were performed in accordance with relevant guidelines and regulations. GBM primary cultures (GBM49, GBM50, GBM57, GBM71, GBM72, GBM73) were grown in cultivation flask in cultivation medium: RPMI 1640 medium (Sigma-Aldrich) supplemented with 15% fetal bovine serum (Gibco, Thermo Fisher Scientific), insulin (100 IU/ml, Eli Lilly) and transferrin (2 mg/ml, Sigma-Aldrich).…”

Role of Circular RNA in Brain Tumor Development

“…It is a glial cell tumor with a poor prognosis and a high mortality rate in adults [ 60 ]. GBM is resistant to apoptosis induction and overexpresses anti-apoptotic proteins [ 61 ]. The propensity to proliferate rapidly has been identified as a characteristic of GB, resulting in therapies followed by a poor clinical evolution [ 62 ].…”

Glioblastoma and MiRNAs

“…It originates from glial cells [ 12 ] and presents a poor prognosis and high lethality in adults [ 7 ]. GB is resistant to the induction of apoptosis, and anti-apoptotic proteins are overexpressed in GB [ 13 ]. The fast proliferative ability has been characterized as one typical feature of GB, leading to treatments and dismal prognosis [ 14 ].…”

“…MiRNAs act as diagnostic biomarkers in brain tumors and are used for tumor treatment [ 12 ]. MiRNAs mediate the suppression of anti-apoptotic and induce apoptosis in cancer cells [ 13 ]. Deregulated levels of cerebrospinal fluid (CSF) in miRNA are associated with glioblastoma.…”