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Background and purpose: Glioblastoma multiforme (GBM) is the most invasive type of cancer which starts inside the brain. GBM cells were found to have similar properties to glioblastoma cancer stem cells. CD44 can be used as a marker of the cancer stem cells in a subset of glioblastoma tumor cells. Recent studies showed that CD44 is involved in developing cancer cells via the protein kinase B (PKB or AKT) signaling pathway. Therefore, this study aimed to investigate the CD44 mRNA silencing effects on the glioblastoma cell cycle via AKT signaling pathway. Experimental approach: To determine CD44 expression in the samples of the patients with GBM, we used the analysis of data extracted from TCGA database. qRT-PCR and western blotting were used to evaluate the expression level of genes and proteins. Different cell cycles were evaluated by DAPI staining and flow cytometry. Findings/Results: Bioinformatics results showed that CD44 expression level in GBM tumor samples is higher than in normal samples. Effects of poly (ethylene imine)-polyethylene glycol (PEI-PEG)-loaded CD44 siRNA in cell cycle showed that CD44 silencing could inhibit cell cycle in G0-G1 phase by more than 20% compared to the negative control (P < 0.05). Furthermore, PEI-PEG-loaded CD44 siRNA reduces the expression of cyclin D1 and CKD-4. According to our findings, this structure also prevented AKT phosphorylation at Thr-308 and Ser-473. Conclusion and implications: Our results suggest that PEI-PEG-loaded CD44 siRNA may attenuate the cell cycle by suppressing AKT signaling pathway. [ABSTRACT FROM AUTHOR]

Objectives: In this study, the inhibitory effects of polyethylene imine glycol (PEI-PEG)/CD44 siRNA nanostructures on the proliferation, invasion, and apoptosis of U87MG GBM cancer cell line, as well as the expression levels of ALDH1, RANKL, and NOTCH1 were evaluated. Methods: In this experimental study, PEI-PEG/CD44 siRNA nanoparticles were synthesized and characterized by atomic force microscopy (AFM), evaluation of size and zeta potential, and Fourier transform infrared (FTIR) spectroscopy. The MTT assay was adopted to evaluate the cytotoxicity of the nanoparticles. The expression levels of target genes were assessed by qRT-PCR. Flow cytometry was used for apoptosis evaluation and Trans well matrigel assay and scratch-migration were employed for investigating the invasion and migration of glioma cells. Results: The size and zeta potential of PEI-PEG were influenced after CD44 siRNA loading. PEI-PEG loaded with CD44 siRNAs resulted in significant inhibition of glioblastoma cell line in the concentration of 60 pmol (P<0.05). In addition, transfection of glioma cells with CD44 siRNA led to significant downregulation of ALDH1, NOTCH1, and RANKL1 (P<0.05). Transfection of this siRNA also resulted in significant suppression of invasion and migration (P<0.05). Conclusions: PEI-PEG could effectively form the polyplex in combination with siRNA, be transfected into the U87MG glioma cancer cell line, and inhibit the proliferation, invasion and migration of glioma cells via suppression of ALDH1 and NOTCH1, as well as RANKL1 expression levels. [ABSTRACT FROM AUTHOR]

Background and Purpose: Glioblastoma multiforme (GBM) is the most invasive type of cancer which starts inside the brain. GBM cells were found to have similar properties to glioblastoma cancer stem cells. CD44 can be used as a marker of the cancer stem cells in a subset of glioblastoma tumor cells. Recent studies showed that CD44 is involved in developing cancer cells via the protein kinase B (PKB or AKT) signaling pathway. Therefore, this study aimed to investigate the CD44 mRNA silencing effects on the glioblastoma cell cycle via AKT signaling pathway., Experimental Approach: To determine CD44 expression in the samples of the patients with GBM, we used the analysis of data extracted from TCGA database. qRT-PCR and western blotting were used to evaluate the expression level of genes and proteins. Different cell cycles were evaluated by DAPI staining and flow cytometry., Findings/results: Bioinformatics results showed that CD44 expression level in GBM tumor samples is higher than in normal samples. Effects of poly (ethylene imine)-polyethylene glycol (PEI-PEG)-loaded CD44 siRNA in cell cycle showed that CD44 silencing could inhibit cell cycle in G0-G1 phase by more than 20% compared to the negative control ( P < 0.05). Furthermore, PEI-PEG-loaded CD44 siRNA reduces the expression of cyclin D1 and CKD-4. According to our findings, this structure also prevented AKT phosphorylation at Thr-308 and Ser-473., Conclusion and Implications: Our results suggest that PEI-PEG-loaded CD44 siRNA may attenuate the cell cycle by suppressing AKT signaling pathway., Competing Interests: All authors declared no conflict of interest in this study., (Copyright: © 2021 Research in Pharmaceutical Sciences.)