Current Molecular Medicine - Online First

To investigate the effect of the SUMOylation inhibitor TAK981 on hydrogen peroxide (H2O2)-induced oxidative damage in human retinal pigment epithelial cells (ARPE-19) and its regulatory mechanism.

An oxidative damage model of ARPE-19 cells induced by H2O2 was established, and 1, 2, and 5 μM TAK981 solutions were administered for intervention respectively. Normal cells were used as the control group. The viability of the cells in each group was detected by the methyl thiazolyl tetrazolium (MTT) method. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in each group of cells were detected by biochemical methods. The levels of IL-1β and TNF-α produced by each group of cells were detected by enzyme-linked immunosorbent assay (ELISA). The protein expression levels of Nrf2, HO-1, NQO-1, Keap1, and Sumo1 in each group of cells were detected by Western blotting. In addition, 2 μM TAK981 and 2 μM TAK981 combined with 10 μM ML385 (an Nrf2 inhibitor) were administered to H2O2-induced ARPE-19 cells, and the levels of SOD and MDA, IL-1β and TNF-αwere detected again.

The viability of the ARPE-19 cells decreased with increasing H2O2 concentration (F=19.158, P<0.001). H2O2 treatment at 350 μM was the concentration at which the cells essentially reached half inhibition (IC50), and the cell oxidative damage model was successfully established. After intervention with TAK981, cell survival increased significantly (F=0.098, P<0.001). The differences between the 2 μM and 5 μM TAK981 groups and the model group were statistically significant (all P<0.01). Compared with those in the normal group, the MDA content in the model group increased, the SOD activity decreased, and the release levels of IL-1β and TNF-α increased (all P<0.01). Compared with those in the model group, the MDA content in the TAK981 group decreased, the SOD activity increased, and the release levels of IL-1β and TNF-α decreased. The differences between the 2 μM and 5 μM TAK981 groups were statistically significant (P<0.05). Compared with those in the normal group, the protein expression levels of Nrf2, HO-1 and NQO-1 in the model group were greater, whereas the protein expression levels of Keap1 and Sumo1 were lower (all P<0.05). Compared with those in the model group, the protein expression levels of Nrf2, HO-1 and NQO-1 in the TAK981-treated group continued to increase, whereas the protein expression levels of Keap1 and Sumo1 continued to decrease. The differences in the 5 μM TAK981 group were statistically significant (P<0.05). In addition, after the combined intervention of TAK981 and ML385 on H2O2-induced cells, compared with the TAK981-only intervention on H2O2-induced cells, the cell viability increased, the MDA content increased, the SOD activity decreased, and the IL-1β and TNF-α release levels increased. The differences were statistically significant (P<0.05).

The SUMOylation inhibitor TAK981 activates the Keap1/Nrf2/ARE signaling pathway, enhances the activity of antioxidant enzymes, and reduces the production of oxidative stress products and inflammatory factors, thereby exerting a protective effect on H2O2-induced oxidative damage in ARPE-19 cells. Therefore, it is suggested that intervention in SUMO regulation can be used as a new therapeutic target in the AMD disease model, in order to delay the development of AMD by reducing the oxidative damage of RPE.

Inflammation is the natural defense mechanism of the body in response to injury, infection, or other stimuli. Excessive or persistent inflammatory responses can lead to the development of inflammatory diseases. Therefore, elucidating the regulatory mechanisms of inflammatory cells is crucial for understanding the pathogenesis of such diseases and devising novel therapeutic approaches. Moreover, miR-144/451 plays an important role in erythroid maturity and tumour development. Herein, we have reviewed the regulatory role of miR-144/451 in inflammation.

Papers on miR-144, miR-451, and inflammation were retrieved from PubMed and Web of Science to be analysed and summarised.

miR-144/451 plays a significant role in modulating inflammatory responses. Pro- and anti-inflammatory gene transcription is regulated by miR-144/451 binding to the 3′ untranslated regions. Studies have shown that miR-451 inhibits the activation of various inflammatory cells, including macrophages, neutrophils, and T lymphocytes, thereby reducing the release of inflammatory mediators. However, miR-144 expression varies in different inflammatory diseases. miR-144 expression is downregulated in macrophages after induction by lipopolysaccharide, cysteine, or Mycobacterium tuberculosis, which promotes the secretion of inflammatory mediators; nonetheless, miR-144-3p overexpression in macrophages can aggravate atherosclerosis. Meanwhile, miR-144 overexpression prevents disruption of the lung endothelial cell barrier, whereas it exacerbates endothelial cell injury in Crohn’s disease.

miR-144/451 may serve as a potential target for the treatment of inflammatory diseases.

Endometriosis (EM) is a gynecological disease characterized by the benign growth of endometrial tissue outside the uterus. Upregulation of neuronally expressed developmentally downregulated 4 (NEDD4) has been reported to accelerate endometrial cancer progression.

We explored whether abnormal expression of NEDD4 is correlated with EM.

Endometrial tissue in patients without endometriosis was used to develop the original generation of endometrial stromal cells (ESCs). Different types of endometrial tissue of patients with endometriosis were used to measure the expression of NEDD4 by immunohistochemistry (IHC) and western blotting. Its biological functions in ESCs were investigated using a cell counting kit-8 assay, fluorescein diacetate (FDA) staining, and Transwell invasion assays. Additionally, its involvement in ferroptosis was assessed by measuring Fe2+, malondialdehyde (MDA), glutathione (GSH), and reactive oxygen species (ROS) levels and the expression of ferroptosis markers.

Compared with normal controls, NEDD4 levels were significantly elevated in the endometrial tissue of patients with EM. Furthermore, NEDD4 expression was higher in the ectopic endometrium than in the eutopic endometrium. NEDD4 knockdown reduced the viability and invasive capacity of ESCs, increased Fe2+, MDA, and ROS levels, and decreased GSH content. Further analysis revealed that NEDD4 knockdown promoted ferroptosis in ESCs by increasing the expression of prostaglandin-endoperoxide synthase 2 (PTGS2). As an E3 ubiquitin ligase, NEDD4 reduced PTGS2 protein levels by accelerating its ubiquitination and subsequent proteasomal degradation.

These findings suggest that inhibiting NEDD4 reduces ESC growth and invasion in EM by regulating PTGS2-dependent ferroptosis.

Currently, Medullary Thyroid Carcinoma (MTC) is considered a kind of rare neuroendocrine tumor, and molecular-targeted drugs have previously been used for MTC treatment.

However, the prognosis of MTC patients is still not significant. In the present work, we aimed to explore the antitumor activity of the molecularly targeted drug anlotinib in combination with radiofrequency ablation on MTC.

The targets of anlotinib were clearly expressed in MTC tissue specimens, and the expression level of these factors was much higher in MTC clinical specimens than in nontumor tissues. At the same time, anlotinib or Radiofrequency Ablation (RFA) showed clear antitumor activity against the MTC cell line TT (TT cells) and the tumor tissue it formed. Anlotinib, in combination with RFA, significantly increased the antitumor activity of RFA.

These results indicated that the combination of anlotinib with radiofrequency ablation could be a promising therapeutic strategy for MTC treatment.

High morbidity, high mortality and poor prognosis of esophageal squamous cell carcinoma (ESCC) highlights the urgent need for novel therapeutic strategies against ESCC. The current study addresses the precise role of M2-like macrophages-derived CCL17 in ESCC progression and to thoroughly elucidate the intrinsic molecular mechanisms.

In this work, for functional experiments, Eca109 cells cultivated in M2-CM were treated with anti-IgG (50 µg/ml) or anti-CCL17 (50 µg/ml) to expound the tumor-promoting effects of M2-like macrophage-derived CCL17 in ESCC. Moreover, for rescue experiments, Eca109 cells were treated with CCL17 (50 ng/ml) and/or CCR4 antagonist AZD2098 (20 µM) to probe whether CCL17 could influence the malignant behaviors including migration, invasion and stemness of ESCC cells via activating CCR4/ERK/PD-L1 pathway.

Markedly enhanced CCL17 secretion was observed in M2-like macrophages. CCL17 bound to CCR4 to activate ERK/PD-L1 signaling. M2-like macrophages-derived CCL17 facilitated ESCC cell migration and invasion and enhanced stemness characteristics of ESCC cells, which were partially reserved by AZD2098 treatment. The tumor-promoting effects of M2-like macrophages-derived CCL17 on ECSS was depended on the activation of CCR4/ERK/PD-L1 pathway.

To conclude, M2-like macrophages-derived CCL17 could facilitate ESCC cell migration and invasion and enhance stemness characteristics of ESCC cells via activating CCR4/ERK/PD-L1 signaling.

Breast cancer is the most prevalent cancer among women and is usually treated with antineoplastic drugs. The present study examines the influence of sodium deoxycholate on the molecular pathways underlying apoptosis, cytotoxicity, and the modulation of PON1 in the MCF-7 breast cancer cell line. Various doses were administered to test the hypothesis that it could potentially affect cancer cells.

The study examined the cytotoxic effect of sodium deoxycholate on MCF-7 cells and human mammary epithelial cells (CRL-4010) using the MTT method to detect its anticancer properties. Subsequently, the efficacy of the active dose on DNA fragmentation and apoptosis was examined using the apoptotic DNA ladder and Western blot methods. Additionally, oxidative stress index and cell migration tests were conducted. Notably, sodium deoxycholate did not cause DNA damage despite demonstrating cytotoxic effects on cells.

The study found that sodium deoxycholate increased the levels of several pro-apoptotic proteins, leading to apoptosis. Moreover, it markedly diminishes the activity of paraoxonase and arylesterase of PON1, which are predictive risk markers for cancer. Furthermore, it was found to delay cell migration in a time-dependent manner.

These findings suggest that sodium deoxycholate exhibits an antimetastatic effect in breast cancer cells, could be a valuable subject for further cancer research.

The aim of this study was to detect the association between the mTOR-STAT3 pathway and focal cortical dysplasia type IIIa (FCD IIIa) in children.

A retrospective review was conducted based on 26 pediatric patients diagnosed with FCD IIIa who underwent surgical intervention. These patients were selected from a cohort of 157 individuals presenting with temporal lobe epilepsy. For comparative analysis, a control group consisting of 5 children who underwent intracranial decompression was established. Immunohistochemistry, immunofluorescence, and western blot techniques were used to assess the expression levels of mTOR, P-mTOR, P-70s6k, STAT3, P-STAT3, and GFAP in brain tissue specimens obtained from the two groups.

The mTOR-STAT3 pathway exhibited activation in the FCD IIIa group (all P < 0.01). Additionally, immunofluorescence analysis revealed that cells positive for P-STAT3 were identified as astrocytes. Moreover, within the FCD IIIa group, there was a marked elevation in the expression of the mTOR-STAT3 pathway in the hippocampus compared to the brain cortex tissue.

The mTOR-STAT3 pathway was demonstrated to be substantially associated with FCD IIIa in pediatric patients. The activation of the mTOR-STAT3 signaling pathway may contribute to the pathogenesis of FCD IIIa in pediatric patients by modulating the proliferation of astrocytes.