Isolation of Cells and Exosomes from Glioblastoma Tissue to Investigate the Effects of Ascorbic Acid on the c-Myc, HIF-1α, and Lnc-SNHG16 Genes.

IF 1.5 Q3 MEDICINE, RESEARCH & EXPERIMENTAL
Masoumeh Eliyasi Dashtaki, Alireza Tabibkhooei, Sepideh Parvizpour, Ramin Soltani, Sorayya Ghasemi
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引用次数: 0

Abstract

Glioblastoma multiforme (GBM) is incurable with routine treatments. Ascorbic acid (Asc) has antioxidant and anti-cancer properties. However, its specific anti-cancer mechanisms are only partially understood. In this study, the effect of Asc on the c-Myc, HIF-1α, and lnc-SNHG16 genes in GBM cells and their exosomes was investigated. Cells isolated from the tissue were characterized by the immunocytochemistry method (GFAP+). The cell-doubling time was determined, and FBS-free medium supplemented with Asc (5 mM) was added to the cells. The extracted exosomes in the cell culture medium were scanned by electron microscopy, Zetasizer, and BCA assay. The expression of lnc-SNHG16 in the exosomes and c-Myc and HIF-1α in the treated and control cells was evaluated by real-time PCR. The interactions between Asc and the c-Myc and HIF-1α proteins were studied using the molecular docking method. The cells showed 90-100% GFAP+ in passage 4, with a cell-doubling time of 4.8 days. Exosomal vesicles measuring 98.25-105.9 were observed. Zetasizer results showed a sharp pick at 90 nm. Protein quantitation showed 3.812 µg/ml protein in the exosomes. Lnc-SNHG16 expression was reduced (P = 0.041), and c-Myc was upregulated (P = 0.002). The expression of HIF-1α was not significant in the treated cells. Also, Asc was able to interact and affect c-Myc and HIF-1α. Asc exerts its effect by reducing lnc-SNHG16 expression in exosomes, upregulating c-Myc in GBM cells, and interacting with HIF-1α and c-Myc. Further research is necessary to achieve a full understanding of these findings.

从胶质母细胞瘤组织中分离细胞和外泌体,研究抗坏血酸对 c-Myc、HIF-1α 和 Lnc-SNHG16 基因的影响。
多形性胶质母细胞瘤(GBM)无法通过常规治疗治愈。抗坏血酸(Asc)具有抗氧化和抗癌特性。然而,人们对其具体的抗癌机制仅有部分了解。本研究调查了 Asc 对 GBM 细胞及其外泌体中 c-Myc、HIF-1α 和 lnc-SNHG16 基因的影响。通过免疫细胞化学方法(GFAP+)对从组织中分离出来的细胞进行表征。测定细胞倍增时间,并向细胞中添加无 FBS、添加 Asc(5 mM)的培养基。用电子显微镜、Zetasizer和BCA检测法扫描细胞培养基中提取的外泌体。实时 PCR 评估了外泌体中 lnc-SNHG16 的表达,以及处理细胞和对照细胞中 c-Myc 和 HIF-1α 的表达。分子对接法研究了Asc与c-Myc和HIF-1α蛋白之间的相互作用。细胞在第4周期显示出90-100%的GFAP+,细胞倍增时间为4.8天。观察到的外泌体囊泡大小为 98.25-105.9。Zetasizer结果显示,在90纳米处有一个尖锐的拾取点。蛋白质定量显示外泌体中含有 3.812 µg/ml 蛋白质。Lnc-SNHG16 表达减少(P = 0.041),c-Myc 上调(P = 0.002)。在处理过的细胞中,HIF-1α的表达并不显著。此外,Asc 还能与 c-Myc 和 HIF-1α 相互影响。Asc通过减少外泌体中lnc-SNHG16的表达、上调GBM细胞中的c-Myc以及与HIF-1α和c-Myc相互作用来发挥其作用。要全面了解这些发现,还需要进一步的研究。
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来源期刊
CiteScore
3.60
自引率
0.00%
发文量
0
期刊介绍: The International Journal of Molecular and Cellular Medicine (IJMCM) is a peer-reviewed, quarterly publication of Cellular and Molecular Biology Research Center (CMBRC), Babol University of Medical Sciences, Babol, Iran. The journal covers all cellular & molecular biology and medicine disciplines such as the genetic basis of disease, biomarker discovery in diagnosis and treatment, genomics and proteomics, bioinformatics, computer applications in human biology, stem cells and tissue engineering, medical biotechnology, nanomedicine, cellular processes related to growth, death and survival, clinical biochemistry, molecular & cellular immunology, molecular and cellular aspects of infectious disease and cancer research. IJMCM is a free access journal. All open access articles published in IJMCM are distributed under the terms of the Creative Commons Attribution CC BY. The journal doesn''t have any submission and article processing charges (APCs).
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