MicroRNA-27a Promotes Oxidative-Induced RPE Cell Death through Targeting FOXO1

Age-related macular degeneration (AMD) is a multifactor disease, which is primarily characterized by retinal pigment epithelium (RPE) cell loss. Since the retina is the most metabolically active tissue, RPE cells are exposed to consistent oxidative environment. So, oxidation-induced RPE cell death has long been considered a contributor to the onset of AMD. Here, we applied a retinal degeneration (RD) rat model induced by blue light-emitting diode (LED) and a cell model constructed by H2O2 stimulus to mimic the prooxidant environment of the retina. We detected that the expression of miR-27a was upregulated and the expression of FOXO1 was downregulated in both models. So, we furtherly investigated the role of miR-27a-FOXO1 axis in RPE in protesting against oxidants. Lentivirus-mediated RNA was injected intravitreally into rats to modulate the miR-27a-FOXO1 axis. Retinal function and histopathological changes were evaluated by electroretinography (ERG) analysis and hematoxylin and eosin (H&E) staining, respectively. Massive photoreceptor and RPE cell death were examined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL). The damage to the retina was aggravated in the FOXO1 gene-knockdown and miR-27a-overexpression groups after exposure to LED but was alleviated in the FOXO1 gene-overexpression or miR-27a-knockdown groups. Dual luciferase assay was used to detect the binding site of miR-27a and FOXO1. Upregulated miR-27a inhibited the expression of FOXO1 by directly binding to the FOXO1 mRNA 3 ′ UTR and decreased the autophagy activity of ARPE-19 cells, resulting in the accumulation of reactive oxygen species (ROS) and decrease of cell viability. The results suggest that miR-27a is a negative regulator of FOXO1. Also, our data emphasize the prominent role of miR-27a/FOXO1 axis in modulating ROS accumulation and cell death in RPE cell model under oxidative stress and influencing the retinal function in the LED-induced RD rat model.

Comparison of the effect of exercise with Kelussia Odaratissma Mozaff on FoxO1 Expression level in cardiac tissue of obese rats

Background and Aim: The FoxO1 gene is an important regulator of cellular metabolism in heart tissue. Therefore, the aim of this study was to investigate the effect of Kelussia extract and exercise on FoxO1 gene expression in vascular tissue. Materials & Methods: In the present study, 30 adult male Wistar rats weighing 180 to 200 g were selected and divided into five groups of control, negative control, one group of obese rats with exercise and two treatment groups with doses of 400 and 800 mg / Kg of Kelussia extract were divided. Molecular tests were performed on the hearts of 30 rats using Real Time RT PCR. Finally, expression test was performed using SPSS software and ANOVA TEST and LSD tests with a significance level of P <0.05. Results: Kelussia extract at a dose of 400 mg / kg can significantly reduce the expression of FoxO1 gene (0.61 ± 0.14 ab) compared to the 800 dose (0.70 ± 0.25 ab) compared to the obese group. Statistically significant was significant (p <0.05). Also, in the group of rats with exercise, we had a decrease in gene expression compared to the obese group (0.54 ± 0.13 b). On the other hand, the results of biochemical tests confirmed the reduction of factors in the hearts of obese rats in rats treated with 400 mg / kg of Kelussia. Discussion and Conclusion: Exercise as well as Kelussia extract can reduce the expression of the FoxO1 gene and, if confirmed in future studies, could be a therapeutic target in heart disease.

The Mutual Inhibition of FoxO1 and SREBP-1c Regulated the Progression of Hepatoblastoma by Regulating Fatty Acid Metabolism

Background. Hepatoblastoma (HB) is the most common liver malignancy in pediatrics, but the treatment for this disease is minimal. This study is aimed at exploring the effect of FoxO1 and SREBP-1c on HB and their mechanism. Methods. FoxO1, SREBP-1c, FASN, ACLY, ACC, and MAGL expressions in tissue samples were detected by RT-qPCR and WB. IHC was utilized to measure FASN content. Overexpression and knockdown of FoxO1 and sSREBP-1c were performed on Huh-6 cells. Cell proliferation, migration, and invasion were examined by CCK8, scratch, and transwell assay. ELISA was performed to test the ATP, FAO, NEFA, and Acetyl-CoA contents. ChIP was used to detect the interaction between SREBP-1c protein and the FoxO1 gene. In vivo tumorigenesis was conducted on mice. The morphology of tumor tissue sections was observed by HE staining. Results. FoxO1 expression was downregulated in HB tissue, while the expressions of SREBP-1c, FASN, ACLY, ACC, and MAGL were upregulated. In Huh-6 cells and mouse tumor tissues, FoxO1 knockdown resulted in increased cell proliferation, migration, and invasion and active fatty acid metabolism. On the contrary, after the knockdown of SREBP-1c, cell proliferation, migration, and invasion were weakened, and fatty acid metabolism was significantly reduced. SREBP-1c interacted with the promoter of the FoxO1 gene. When FoxO1 was knocked down, the tumor tissue was more closely packed. After the knockdown of the SREBP-1c gene, the structure of tumor cells was deformed. Conclusion. FoxO1 and SREBP-1c inhibited each other in HB, leading to the increase of intracellular fatty acid metabolism, and ultimately facilitated the development of HB.

Genetic and Functional Evaluation of the Role of FOXO1 in Antituberculosis Drug-Induced Hepatotoxicity

Background. The accumulation of the hepatotoxic substance protoporphyrin IX (PPIX) induced by aminolevulinate synthase 1 (ALAS1) activation is one of the important mechanisms of antituberculosis drug-induced hepatotoxicity (ATDH). Forkhead box protein O1 (FOXO1) may activate ALAS1 transcription. However, little is known about their roles in ATDH; we performed a study to determine the association between polymorphisms in the two genes and ATDH susceptibility. Then, we verified this possible association by cellular functional experiments. Materials and Methods. Tag single-nucleotide polymorphisms (TagSNPs) in the two genes were genotyped in 746 tuberculosis patients. The frequencies of the alleles, genotypes, genetic models, and haplotype distribution of the variants were compared between the case and control groups. L-02 cells and HepG2 cells were incubated with the indicated concentration of isoniazid (INH) and rifampicin (RIF) for the desired times, and then the expression levels of ALAS1 and FOXO1 mRNAs and proteins were detected. HepG2 cells were transiently transfected with FOXO1 siRNA to observe the effect of changes in the FOXO1 expression on the cell survival rate and ALAS1 expression. Results. The C allele at rs2755237 and the T allele at rs4435111 in the FOXO1 gene were associated with a decreased risk of ATDH. The expression of ALAS1 in both L-02 cells and HepG2 cells was increased by the coadministration of INH/RIF (600/200 μM) for 24 h. Although FOXO1 expression was reduced slightly by the same treatment, its content in the nucleus was significantly increased. However, the cell survival rate and ALAS1 expression level were not significantly altered by the downregulation of FOXO1 in HepG2 cells. Conclusions. Variants of the rs4435111 and rs2755237 loci in the FOXO1 gene were associated with susceptibility to ATDH. Coadministration of INH/RIF promoted the transfer of FOXO1 from the cytoplasm to the nucleus, but the functional significance of its nuclear translocation requires further verification.

Adult Primary Pineal Alveolar Rhabdomyosarcoma With FOXO1 Gene Rearrangement and OLIG2 Expression: a Case Report and the Literature Review

Introduction: Alveolar rhabdomyosarcoma (ARMS) is a common malignant soft tissue tumor in child and adolescents. When diagnosed in adults, it is an aggressive, often fatal disease. Intracranial ARMS in adults is rare, especially in the pineal region. Case presentation: A 36-year-old Chinese man presented with a 3-month history of dizziness and 1-month history of headache and unsteady walking. Magnetic resonance imaging of the brain revealed a pineal region lesion with obstructive hydrocephalus. He received ventricular-abdominal shunt and endoscopic-assisted pineal lesion resection.The tumor appears as a solid sheet-like growth of medium-sized round or oval cells with map-like necrosis and some rhabdomyoblasts. Immunohistochemical test revealed that tumor cells were diffusely positive for desmin, myogenin, MyoD1, ALK and CD56. Notably, the tumors were diffusely positive for OLIG2. Fluorescence in situ hybridization confirmed the FOXO-1 gene rearrangement. The final diagnosis of the present case is ARMS of the pineal gland. Conclusions: We presented an extremely rare case of primary ARMS in adult pineal region particularly with expression of OLIG2 and confirmed by PAX3/7-FOXO1 fusion gene detection.

FOXO1 gene involvement in a non-rhabdomyosarcomatous neoplasm

Myoepithelial neoplasms of soft tissue are rare tumors with clinical, morphological, immunohistochemical, and genetic heterogeneity. The morphological spectrum of these tumors is broad, and the diagnosis often requires immunostaining to confirm myoepithelial differentiation. Rarely, tumors show a morphology that is typical for myoepithelial neoplasms, while the immunophenotype fails to confirm myoepithelial differentiation. For such lesions, the term “myoepithelioma-like” tumor was introduced. Recently, two cases of myoepithelioma-like tumors of the hands and one case of the foot were described with previously never reported OGT-FOXO gene fusions. Here, we report a 50-year-old woman, with a myoepithelial-like tumor localized in the soft tissue of the forearm and carrying a OGT-FOXO1 fusion gene. Our findings extend the spectrum of mesenchymal tumors involving members of the FOXO family of transcription factors and point to the existence of a family of soft tissue tumors that carry the gene fusion of the OGT-FOXO family.

Prevalence of FOXO1 gene abnormalities in a group of round-cell rhabdomyosarcomas with alveolar morphology

Rhabdomyosarcomas (RMS) are group of soft tissue malignant tumours predominantly childhood. Alveolar rhabdomyosarcoma (aRMS) is the second most common variant of RMS. The majority of aRMSs display a translocations of FOXO1 gene. Such tumours are aggressive, metastasize early and are associated with a worse prognosis for the patient. However, some aRMS cases are rhabdomyosarcomas without classic chromosomal rearrangements. These tumors also have alveolar morphology, but neoplastic cells lack the FOXO1 gene translocation. Such fusion-negative round-cell RMSs behave clinically differently and have a better prognosis. The aim of the present study was to assess the prevalence of FOXO1 gene rearrangements in the group of round cell rhabdomyosarcomas with alveolar morphology. This study is supported by the Independent Ethics Committee and approved by the Academic Council of the Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology. The study group consisted of 250 formalin-fixed paraffin-embedded samples from patients with RMS. The cytogenetic study was performed by fluorescence in situ hybridization with a locus-specific identifier (LSI) for FOXO1 (13q14). The PAX3-FOXO1 (COSF247) and PAX7-FOXO1 (COSF287) fusion transcripts was detected by RT-PCR. In the study group 1 (аRMS), the rearrangement of PAX3/7-FOXO1 was detected in 44% of cases, in 32% of cases was detected a combined rearrangement with amplification of the 3' FOXO1. In one case, the amplification of the 3' end of the FOXO1 gene was detected without rearrangement; this sample was sent for additional PCR study, as a result of which the chimeric PAX3-FOXO1 transcript was detected. In 22% cases cytogenetic abnormalities were not found. has not been identified. In group 2 (embryonal RMS) we did not detect translocation. The group of round-cell PMCs is heterogeneous and is represented by three variants of genetic events that determine the disease prognosis. At the same time, FOXO1 gene abnormalities are not found in the RMS group with non-alveolar morphology.

FoxO1, a Potential Therapeutic Target, Regulates Mitochondrial Dysfunction and Pyroptosis in LPS-Induced Human Cervical Cancer Cells

Background: Chronic inflammation plays an important role in the development of cervical cancer. Studies have demonstrated that transcription factors forkhead box protein O1 (FoxO1) have been reported to play important roles in various cancers. Aim: The purpose of this study was to investigate the effect of FoxO1 gene on lipopolysaccharide (LPS)-induced inflammation and intracellular pyroptosis in the development and progression of human cervical cancer cells (SiHa).Methods: In this study, FoxO1 expression was examined using real-time polymerase chain reaction (PCR), western blotting and immunohistochemical staining. SiHa cells migration and proliferation was detected using the transwell assay and 3H‑TdR assay. Mitochondrial function was assessed based on reactive oxygen species (ROS) generation and changes in the mitochondrial membrane potential (ΔΨm). Results: Our study demonstrated that LPS inhibited FoxO1 gene expression, and the silence of FoxO1 gene caused the accumulation of ROS, decreases in the ΔΨm and mitochondrial morphology change). However, either overexpression of FoxO1 or metformin could reverse the LPS-induced mitochondrial dysfunction, cell pyroptosis, migration and proliferation.Conclusions: Our study indicated that FoxO1 as a potential therapeutic target to cure against LPS-induced human cervical cancer in a mitochondria-dependent manner.