Caspase‑dependent apoptotic death by gadolinium chloride (GdCl3) via reactive oxygen species production and MAPK signaling in rat C6 glioma cells

Yang

et al. 2021

In Vivo

Background/Aim: Magnetic resonance imaging (MRI) is a technique for evaluating patients with primary and metastatic tumors. The contrast agents improve the diagnostic accuracy of MRI. Large quantities of a contrast agent must be administrated into the patient to obtain useful images, which leads to cell injury. Gadolinium has been reported to cause central lobular necrosis of the liver and nephrogenic systemic fibrosis. However, the toxicity caused on brain tissue is uncertain. Materials and Methods: This study mainly aimed on the in vitro study of high concentration (2 and 5-fold of normal concentration) gadolinium-based contrast agents (GBCAs), gadodiamide (Omniscan ® ), on normal brain glial SVG P12 cells. MTT assay, DAPI staining, immunofluorescent staining, LysoTracker Red staining, and western blotting analysis were applied on the cells. Results: The viability of gadodiamide (1.3, 2.6, 5.2, 13 and 26 mM)-treated SVG P12 cells was significantly reduced after 24 h of incubation. Gadodiamide caused significant autophagic flux at 2.6, 5.2 and 13.0 mM as seen by acridine orange (AO) staining, LC-3-GFP and LysoTracker Red staining. The expression levels of autophagy-related proteins such as beclin-1, ATG-5, ATG-14 and LC-3 II were up-regulated after 24 h of gadodiamide incubation. Autophagy inhibitors including 3-methyladenine (3-MA), chloroquine (CQ) and bafilomycin A1 (Baf) significantly alleviated the autophagic cell death effect of gadodiamide on normal brain glial SVG P12 cells. Gadodiamide induced significant apoptotic effects at 5.2 mM and 13.0 mM as seen by DAPI staining and the pan-caspase inhibitor significantly alleviated the apoptotic effect. Gadodiamide at 5.2 mM and 13.0 mM inhibited antiapoptotic protein expression levels of Bcl-2 and Bcl-XL, while promoted pro-apoptotic protein expression levels of Bax, BAD, cytochrome c, Apaf-1, cleaved-caspase-9 and cleaved-caspase-3. Conclusion: Normal brain glial SVG P12 cells treated with high concentrations of gadodiamide can undergo autophagy and apoptosis.Magnetic resonance imaging (MRI) is a non-invasive technique used to produce detailed images of internal structure in the human body and diagnose medical conditions. The density of adjacent organs is similar. Therefore, it is impossible to distinguish the shape, location 2621 This article is freely accessible online.

“…Cell viability by MTT assay. The cell viability was assessed using MTT (thiazolyl blue tetrazolium bromide) colorimetric assay (20). SVG p12 cells were seeded in a 96-well culture dish.…”

Section: Methodsmentioning

confidence: 99%

In VitroToxicological Assessment of Gadodiamide in Normal Brain SVG P12 Cells

Yang

et al. 2021

In Vivo

“…This indicates that TGF is not only a hot topic in cancer research but also could be a target for glioma therapy. Numerous studies have shown that cell migration and neuronal migration are associated with local infiltration, distant metastasis, and the recurrence of glioma (17,18), and the MAPK signaling pathway is a key pathway of cellular proliferation and apoptosis (19). All related functions and signaling pathways indicate that these changes are related to glioma cell proliferation, migration, and apoptosis.…”

Section: Discussionmentioning

confidence: 99%

Establishment of a prognostic-related microRNAs risk model for glioma by bioinformatics analysis

Wang

Zhang

et al. 2021

Ann Transl Med

Background: To explore the specific prognosis related microRNAs (miRNAs) of glioma.Methods: The miRNA-Seq data and clinical information of glioma patients were downloaded from the TCGA (510 cases) and GEO (GSE112009, 25 cases) database. LASSO & COX regression was used to develop a miRNA-based model for predicting patient survival in the training set (n=255), to carry out glioma prognostic related miRNAs screening, and to construct a linear risk model based on the expression profiles of seven miRNAs. COX regression analysis was used to determine whether the miRNAs risk model was an independent prognostic factor.Results: Seven survival-related miRNAs (miR-140-5p, miR-145-5p, miR-148a-3p, miR-183-5p, miR-222-3p, miR-223-3p, and miR-374a-5p) were identified in the training set. This showed that the overall survival time of the high-risk group was significantly lower than that of the low-risk group in the training set, prediction set, and validation set (P<0.05). Further analysis revealed that age and Karnofsky score both affected the risk of glioma. By crossing seven potential target genes of microRNAs, 620 effective target genes were obtained and GO analysis showed that these were related to the positive regulation of cell migration, neuron migration, and the response of transforming growth factor, and KEGG analysis showed they were related to the TGF-beta signaling pathway, MAPK signaling, and AGE-RAGE signaling pathway in diabetic complications.Conclusions: Seven miRNAs which regulate target genes to participate in related signaling pathways and lead to a poor prognosis were identified as biomarkers of glioma.

“…HEK 293 cells (0.8×10 4 cells/well) were grown in 96 well plates for 24 h and treated with different concentrations (0, 25, 50, 100, and 200 mgI/ml) of iopromide for either 24 or 48 h. Cell morphology was examined to assess apoptosis or autophagy using a light microscope at 200× magnification (Leica Microsystems GmbH, Wetzlar, Germany). Cell viability was assessed using the MTT assay (thiazolyl blue tetrazolium bromide) as described previously (36). Briefly, the cells were mixed with MTT for 4 h. The absorbance at 570 nm was measured using a SpectraMax iD3 multimode microplate reader (Molecular Devices Ltd., San Jose, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

“…To determine the alterations in caspase enzymatic activity, FAM FLICA ® Caspases-3/7 Assay Kit and FAM FLICA ® Caspases-9 Assay Kit (MyBioSource, San Diego, CA, USA) were used according to the manufacturer's instructions. Briefly, HEK 293 cells were seeded into 6-well plates at a density of 1×10 6 cells/well for 24 h and treated with iopromide (50 mgI/ml or 100 mgI/ml) for another 48 h. Samples were assessed as described previously (36,37) and analyzed using the NucleoCounter ® NC-3000™ advanced image cytometer (ChemoMetec A/S).…”

Section: Methodsmentioning

confidence: 99%

High Concentration of Iopromide Induces Apoptosis and Autophagy in Human Embryonic Kidney CellsviaActivating a ROS-dependent Cellular Stress Pathway

Tsai¹,

Yang

et al. 2021

In Vivo

Background/Aim: The use of iodinated contrast media may impair renal function. However, no report has addressed the nephrotoxicity of high doses of iodinated contrast media in normal kidney cells and its associated molecular mechanisms. Materials and Methods: Cell proliferation was assessed using the MTT assay. Cell death was evaluated through examining the morphological changes and TUNEL assay. Autophagy was detected through acridine orange staining and lysotracker staining. Reactive oxygen species production and AKT kinase activity were examined. Results: Iopromide induced cell death and triggered apoptosis and autophagy in HEK 293 cells. Cell viability was significantly restored in the presence of a pan-caspase inhibitor or a ROS scavenger, N-acetyl-L-cysteine. AKT kinase activity was found to be reduced in iopromide-treated HEK 293 cells. Conclusion: High concentrations of iopromide induce cell damage, apoptosis, and autophagy through down-regulating AKT and ROS-activated cellular stress pathways in HEK 293 cells.Roentgen discovered X-rays at the end of 19 th century, which was followed by the introduction of iodine-containing contrast media (ICCM) at the beginning of the 20 th century that opened up a new field of research in medical science (1-4). Iodine-containing contrast agents are administrated into the human body through veins and used for radiological diagnostic examinations of the urinary system, heart, cerebrospinal system, and blood vessels in various parts of the body. ICCM enhances the contrast between normal tissue and lesions, provides information on the morphologic features of the lesions, and improves the sensitivity of the examination and the accuracy of diagnosis. The imaging enhancement effect of the contrast agent makes many tissues and cell lesions visible (5-8). The main pharmacological mechanism is that the iodine molecules in the iodinecontaining contrast agent absorb X-ray energy, which makes the iodine molecules exert a special X-ray shielding effect. Therefore, a white high-density image appears on the X-ray 3221 This article is freely accessible online.