Simultaneous Treatment with P53 Overexpression and Interferon γ Exerts a Dramatic Increase in Apoptosis Induction of U87 Cells

Abstract

Background: Gliomas possess low immunogenicity, which is an inevitable hinder in front of cancer immunotherapy. Different interferons (IFNs) may proceed apoptosis instead in p53-dependent or independent pathways. P53 induces the anti-inflammatory programmed cell death in cancer cells; on the other hand, IFN gamma (IFNγ) is a modulatory/pro-inflammatory cytokine. There are contradictory reports of whether this cytokine can possess an anti- or pro-cancerous impact on tumors. Hence, we aimed to investigate the possible cooperative apoptotic effect of the P53 and IFNγ over expressions on the U87 glioblastoma cell line. Materials and Methods: The P53 expressing vector was amplified by Escherichia coli BL21. This vector was confirmed by the aid of sequencing. At the next step, U87 cells were transfected using lipofectamine. Cells were treated with P53 vector and/or IFNγ. The type of cellular death investigated by flow cytometry and the expression level of cleaved caspase-3 protein was also precisely demonstrated by western blotting. Results: Sequencing results revealed that inserted P53 was identical with human P53. Western blot results revealed that both IFNγ and P53 overexpression could up-regulate cleaved caspase-3 protein expression in this cell line. Interestingly, flow cytometry data determined that concurrent treatment with P53 exogenous overexpression and IFNγ induces about 70% apoptosis in U87; more than the sum of cell death occurs after IFNγ or P53 overexpression alone (~18%+21%=39%). Conclusion: The present study results showed that p53-overexpression and IFNγ could ultimately induce up-regulation of the caspase-3 and ultimately significant apoptosis increasing in the U87 cell line. Although IFNγ is believed to be a pro-inflammatory cytokine and P53 is an anti-inflammatory agent, our results demonstrated that they could act synergistically to induce apoptosis in U87 cells. [GMJ.2021;10:e2270]

P53 status, and G2/M cell cycle arrest, are determining factors in cell-death induction mediated by ELF-EMF in glioblastoma

“…Our results answered some parts of this question, as we can see, the apoptosis rate induced in the U87 cells after ELF-EMF exposure is much more than in the U251 cells. It is shown that overexpression of P53 in the U87 cell line after exposure may be the main reason for apoptosis induction by radiation 51 , 52 , as in the U251 cell line, P53 is mutated it is not out mind to expect that the underlying and P53 -dependent pathways which may result in apoptosis after treatment, do not function correctly 13 . The results obtained for the MCF-7 (with wild type P53) or U251-P53 (U251 cells that received vector overexpressing P53 ) also confirmed the role of this gene in the apoptosis induction by ELF-EMF in GBM cells.…”

“…To assess the P53 role in apoptosis induction by ELF-EMF, pCMV-Neo-Bam-H1-p53 plasmid was used. This plasmid was kindly provided by Dr. Michael Resnick (NIEHS, NIH), by the aid of BamHI restriction enzyme and cloning, cDNA of P53 was inserted into the pCMV-NeoBam vector from Addgene (Cambridge, Massachusetts, USA) (reference 13 shows the vector map). U251 cells were transfected with 100 ng/μl concentration of this plasmid (abbreviated as U251-P53), by Lipofectamine®2000.…”

Effects of Dimethyl Fumarate on the Karnofsky Performance Status and Serum S100β Level in Newly Glioblastoma Patients: A Randomized, Phase-II, Placebo, Triple Blinded, Controlled Trial

“…Due to the rapidly evolving nature of GBM and the high risk of resistance to new drugs, the development of new drugs is not considered efficient [6,7]. Therefore, identifying new applications for existing drugs with known pharmacokinetics and safety characteristics has gained a lot of interest [8,9]. One previously used drug with a good efficacy profile is dimethyl fumarate (DMF).…”