Ketamine induces reactive oxygen species and enhances autophagy in SV‐HUC‐1 human uroepithelial cells

Abstract

This study was aimed at exploring the underlying mechanisms of ketamine in the SV-40 immortalized human ureteral epithelial (SV-HUC-1) cells. The viability and apoptosis of SV-HUC-1 cells treated with 0.01, 0.1, and 1 mM ketamine were respectively detected via cell counting kit-8 (CCK-8) assay and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) staining. Reactive oxygen species (ROS) level was measured through ROS probe staining. Apoptosis-related proteins (B-cell lymphoma 2 [Bcl-2] and Bax) and autophagy-associated proteins (light chain 3-I [LC3-I] and LC3-II) were determined by western blot or immunofluorescent assay. Additionally, transmission electron microscopy (TEM) was used to evaluate the formation of autophagosomes. After cotreatment of 3-methyladenine (3-MA) or N-acetyl-l-cysteine (NAC), the biological functions of SV-HUC-1 cells were analyzed to determine the association of ROS with cell viability and autophagy. CCK-8 assay and TUNEL staining indicated that ketamine effectively decreased the viability of SV-HUC-1 cells and accelerated apoptosis of SV-HUC-1 cells through regulating the expression level of IKBα (phospho), nuclear factor кB (P65), Bcl-2, and Bax proteins. Enhanced ROS production was also confirmed in ketamine-treated SV-HUC-1 cells treated with ketamine. Ketamine-induced autophagosomes in SV-HUC-1 cells were observed by means of TEM, and increased levels of LC3 II/I ratio and Beclin 1 were examined through western blot and immunofluorescent assay. Furthermore, ketamine exerted effects on SV-HUC-1 cells in a dose-dependent and time-dependent manner. Additionally, cotreatment of NAC with 3-MA significantly attenuated the ROS level and suppressed the cell autophagy. Ketamine promoted SV-HUC-1 cell autophagy and impaired the cell viability of SV-HUC-1 cells by inducing ROS.

<p>LncRNA CDKN2B-AS1 Promotes Cell Viability, Migration, and Invasion of Hepatocellular Carcinoma via Sponging miR-424-5p</p>

“…[6][7][8][9] Some lncRNAs have been reported to be closely related to liver cancer pathology, progression, prognosis, and the maintenance of cancer stem celllike properties. 10,11 For example, LINC00668 up-regulation accelerates cell proliferation, migration, and invasion of HCC via targeting miR-532-5p/YY1 axis; 12 HAGLROS is highexpressed in HCC and its high expression is correlated with the progression of HCC by affecting cell proliferation, apoptosis, and autophagy via regulating miR-5095/ATG12 axis; 13 HCC patients with high-expressed lncRNA PDZD7 tend to show a poorer prognosis; PDZD7 promotes stemness properties and inhibits chemosensitivity of HCC via regulating the miR-101/EZH2/ATOH8 pathway. 14 lncRNAs closely associated with HCC should be further investigated for discovering novel promising biomarkers and potential targets for HCC treatment.…”

What urologists need to know about ketamine‐induced uropathy: A systematic review

“…It is still not clear if inflammation is the result of KC or inducing it. However, a number of studies speculated that the presence of ketamine in urine might be the trigger for bladder inflammation and damage 39‐41 . First of all, even after a single dose administration, ketamine and its metabolites are detected in urine up to 14 days, leading to continuous bladder exposure 42 .…”

“…Baker et al showed that ketamine‐induced toxicity to urothelial cells by high and sustained intracellular calcium ion concentration, which ultimately led to apoptosis 40 . This damage acts in a dose and time‐dependent manner, stimulating mitochondrial‐dependent and endoplasmic reticulum stress apoptosis and autophagy 41,43‐46 . Chuang et al showed that ketamine and norketamine levels were much higher in the 28‐day treated group compared to the 14‐day group.…”

MiR-185-3p mimic promotes the chemosensitivity of CRC cells via AQP5

“…Drug resistance remains an obstacle in cancer treatment. Though multiple mechanisms underlying drug resistance in cancers, such as improved DNA repair capacity, 1 defects in the DNA damage response, 2,3 anti-apoptosis, 4 pro-migration, 5 pro-proliferation effects, 6 autophagy, 7 tumor-associated compensatory pathways, 8 and changes in drug transport, 9 have been proposed, the exact mechanism of drug resistance in cancer is highly complex and has not been fully understood yet. Therefore, discoveries of new drug sensitivity-associated targets and effective drug combinations are considered as effective and promising strategies for improving the survival rate of cancer patients.…”