FOXA1 in prostate cancer / 亚洲男科学杂志(英文版)

Most prostate cancers initially respond to androgen deprivation therapy (ADT). With the long-term application of ADT, localized prostate cancer will progress to castration-resistant prostate cancer (CRPC), metastatic CRPC (mCRPC), and neuroendocrine prostate cancer (NEPC), and the transcriptional network shifted. Forkhead box protein A1 (FOXA1) may play a key role in this process through multiple mechanisms. To better understand the role of FOXA1 in prostate cancer, we review the interplay among FOXA1-targeted genes, modulators of FOXA1, and FOXA1 with a particular emphasis on androgen receptor (AR) function. Furthermore, we discuss the distinct role of FOXA1 mutations in prostate cancer and clinical significance of FOXA1. We summarize possible regulation pathways of FOXA1 in different stages of prostate cancer. We focus on links between FOXA1 and AR, which may play different roles in various types of prostate cancer. Finally, we discuss FOXA1 mutation and its clinical significance in prostate cancer. FOXA1 regulates the development of prostate cancer through various pathways, and it could be a biomarker for mCRPC and NEPC. Future efforts need to focus on mechanisms underlying mutation of FOXA1 in advanced prostate cancer. We believe that FOXA1 would be a prognostic marker and therapeutic target in prostate cancer.

Emodin reduces the injury of glomerular mesangial cells in lupus nephritis by targeting forkhead protein K2 through miR-96-5p / 中国临床药理学与治疗学

AIM: To investigate the injury of emodin (EMO) in reduce of glomerular mesangial cells (MCs) in lupus nephritis by targeting forkhead protein K2 (FOXK2) through miR-96-5p. METHODS: The contents of 24 h urine protein, serum urea nitrogen (BUN) and serum creatinine (Scr) in MRL / faslpr mice (lupus nephritis group) and MRL / MPJ mice (control group) were detected. MCs were separated, purified and divided into: MCs group (MCs without any treatment), L-EMO group (MCs treated with 10 μmol/L Emodin), M-EMO group (MCs treated with 25 μmol / L Emodin), H-EMO group (MCs treated with 50 μmol / L Emodin), H-EMO + miR-96-5p-NC group (MCs treated with 50 μmol / L Emodin and transfected with miR-96-5p-NC), and H-EMO + miR-96-5p-minic group (MCs treated with 50 μmol/ L Emodin and transfected with miR-96-5p-minic). Double luciferase report experiment was used to verify the targeting relationship between miR-96-5p and FOXK2. The real-time quantitative fluorescent polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-96-5p. Western blot was used to detect the expression of FOXK2 and apoptosis related proteins. The enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of inflammatory factors in MCs. cell count kit 8 (CCK-8) was used to determine the activity of MCs. Annexin-V FITC/PI double staining was used to detect apoptosis of MCs. RESULTS: Compared with the control group, 24 h urinary protein content, serum BUN and Scr levels in the lupus nephritis group were significantly increased (P< 0.05). Compared with the MCs group, the miR-96-5p expression, interleukin1β (IL-1β), interleukin6 (IL-6), tumor necrosis factor-α (TNF-α), A450 value and B-lymphoblastoma-2 (Bcl-2) protein in the L-EMO group, M-EMO group and H-EMO group were significantly decreased (P<0.05), the FOXK2 level, cell apoptosis rate, Bcl-2 related X gene (Bax), aspartate specific cysteine proteinase-3 (cleaved Caspase-3) protein levels were significantly increased, respectively (P<0.05), the effect of Emodin was dose-dependent. Compared with the H-EMO group and H-EMO+miR-96-5p-NC group, H-EMO+miR-96-5p-minic group obviously increased the miR-96-5p expression, inflammatory factor levels, A450 value and Bcl-2 protein level (P<0.05), and obviously decreased FOXK2 level and cell apoptosis rate (P< 0.05). CONCLUSION: EMO can reduce the injury of lupus nephritis MCs by down-regulating miR-96-5p and then up-regulating FOXK2.

FOXO1-miR-506 axis promotes chemosensitivity to temozolomide and suppresses invasiveness in glioblastoma through a feedback loop of FOXO1/miR-506/ETS1/FOXO1 / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

To explore the role of forkhead box protein O1 (FOXO1) in the progression of glioblastoma multiforme (GBM) and related drug resistance, we deciphered the roles of FOXO1 and miR-506 in proliferation, apoptosis, migration, invasion, autophagy, and temozolomide (TMZ) sensitivity in the U251 cell line using in vitro and in vivo experiments. Cell viability was tested by a cell counting kit-8 (CCK8) kit; migration and invasion were checked by the scratching assay; apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining and flow cytometry. The construction of plasmids and dual-luciferase reporter experiment were carried out to find the interaction site between FOXO1 and miR-506. Immunohistochemistry was done to check the protein level in tumors after the in vivo experiment. We found that the FOXO1-miR-506 axis suppresses GBM cell invasion and migration and promotes GBM chemosensitivity to TMZ, which was mediated by autophagy. FOXO1 upregulates miR-506 by binding to its promoter to enhance transcriptional activation. MiR-506 could downregulate E26 transformation-specific 1 (ETS1) expression by targeting its 3'-untranslated region (UTR). Interestingly, ETS1 promoted FOXO1 translocation from the nucleus to the cytosol and further suppressed the FOXO1-miR-506 axis in GBM cells. Consistently, both miR-506 inhibition and ETS1 overexpression could rescue FOXO1 overactivation-mediated TMZ chemosensitivity in mouse models. Our study demonstrated a negative feedback loop of FOXO1/miR-506/ETS1/FOXO1 in GBM in regulating invasiveness and chemosensitivity. Thus, the above axis might be a promising therapeutic target for GBM.

Mechanism of Momordica charantia Extract in Regulating Gluconeogenesis in ZDF Diabetes Rats / 中国实验方剂学杂志

ObjectiveTo observe the effect of Momordica charantia extract （MCE） on the gluconeogenesis signaling pathway in diabetes rats. MethodMale Zucker Diabetic Fatty （ZDF） rats aged 5-6 weeks were randomly divided into a model group and an MCE group （administered MCE at a dose of 0.40 g·kg-1 by gavage）. Additionally， seven healthy male ZDF （fa/+） rats were assigned to the normal group and received administration once daily for six consecutive weeks. During the experiment， the general condition of the rats was observed， and body weight was recorded. Fasting blood glucose and random blood glucose levels were measured in the 1st， 3rd， and 5th weeks. In the 6th week， an oral glucose tolerance test （OGTT） was conducted， and serum levels of triglycerides （TG）， free fatty acid （FFA）， total cholesterol （TC）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） were measured. Hematoxylin-eosin （HE） staining was performed to examine liver morphology， periodic acid-Schiff （PAS） staining was used to assess hepatic glycogen storage， and Real-time polymerase chain reaction （PCR） was employed to measure the mRNA expression of phosphoenolpyruvate carboxykinase （PEPCK） and glucose-6-phosphatase （G6Pase） in the liver. Western blot analysis was conducted to measure the phosphorylation level of forkhead box protein O1 （FoxO1） and the protein expression of PEPCK and G6Pase in the liver. ResultCompared with the model group， the MCE group showed significant improvements in body weight， fasting blood glucose， random blood glucose， and glucose tolerance （P<0.05， P<0.01） and reduced serum levels of FFA， TC， and TG （P<0.05， P<0.01）. There were no significant differences in ALT and AST between the two groups. In the MCE group， the HE staining revealed more orderly liver cell arrangement and reduced hepatic steatosis and the PAS staining showed increased hepatic glycogen storage. The protein expression of p-FoxO1 in the liver was significantly elevated （P<0.01）， while there was no significant difference in FoxO1 protein expression. The mRNA and protein expression of PEPCK and G6Pase significantly decreased （P<0.05）. ConclusionMCE exhibits glucose-lowering and lipid-lowering effects， improves glucose tolerance， and enhances hepatic glycogen storage. These effects may be attributed to the upregulation of p-FoxO1， leading to the inhibition of PEPCK and G6Pase expression and the regulation of gluconeogenesis-related processes.

Liraglutide improves bone mineral density in osteoporotic rats by mediating O3a/Wnt signaling / 中华内分泌外科杂志

Objective:To study the effect of liraglutide on the forkhead box O3a (forkhead box O3a, FoxO3a) /Wnt signaling pathway and vertebral bone density in osteoporotic rats.Methods:Female Sprague-dawley rats were divided into sham operation group, model group, and liraglutide intervention group according to the random number table method, with 10 rats in each group. Both the model group and the intervention group used bilateral oophorectomy to establish an osteoporosis model. The intervention group was injected subcutaneously with liraglutide every day, and the sham operation group and the model group were given an equal volume of normal saline. After 12 weeks of continuous administration, the bone mineral density and bone biomechanics were measured, the serum osteoprotegerin, anti-tartrate acid phosphatase, and osteocalcin levels were detected by enzyme-linked immunosorbent assay, and the O3a/Wnt pathway was detected by Real-time PCR technology. The expression level of mRNA was detected by Western blot to detect the expression level of related proteins in the O3a/Wnt pathway.Results:The bone mineral density levels of L4,5 (0.33±0.04 vs 0.18±0.03) and left and right femurs (0.37±0.05 vs 0.23±0.04 0.35±0.04 vs 0.24±0.03) of the successfully modeled rats were significantly lower than those of the sham operation group ( P<0.05) . After 12 weeks of treatment, the differences in the maximum bone load, three-point bending stress, bone density and elastic modulus of the three groups of rats were statistically significant. Among them, the sham operation group had the highest level of each index (36.53±5.23, 154.13±6.27, 0.34±0.04, 3 102.34±160.44) , followed by the intervention group (19.37±4.32, 141.54±6.58, 0.18±0.03, 2 270.18±145.53) and the model group in turn (26.17±4.68, 147.56±5.84, 0.28±0.03, 2 804.24±140.42) ( P<0.05) . There were statistically significant differences in the levels of serum osteoprotegerin, anti-tartrate acid phosphatase and osteocalcin among the three groups. Among them, the osteoprotegerin (Sham operation group vs model group vs intervention group: 7.53±0.63 vs 4.57±0.42 vs 6.15±0.61) of the sham operation group was significantly higher than that of the intervention group and the model group. The anti-tartrate acid phosphatase (Sham operation group vs model group vs intervention group: 14.34±2.87 vs 19.53±3.52 vs 15.96±3.14) and osteocalcin levels (Sham operation group vs model group vs intervention group: 0.84±0.09 vs 1.13± 0.12 vs 0.95± 0.08) of rats The factor was significantly lower than that of the intervention group and model group ( P<0.05) . The mRNA and protein expression levels of FoxO3a, Wnt2, and β-catenin in the three groups of rats were significantly different. Among them, Wnt2 and β-catenin in the sham operation group were significantly higher than the intervention group and model group, and FoxO3a was significantly lower than the intervention group and model Group ( P<0.05) . Conclusion:Liraglutide can increase bone density and improve bone biomechanics by activating O3a/Wnt signal, thereby effectively treating osteoporosis.

The expression of FOXO3 in pancreatic cancer and its effects on pancreatic cancer cells / 中华肝胆外科杂志

Objective:To investigate the expression of forkhead box protein O3(FOXO3) in pancreatic cancer and its effect on the motility and proliferation of pancreatic cancer cells.Methods:The FOXO3 expression in pancreatic cancer and adjacent tissues was retrieved from LinkedOmics database. Western blotting and real-time quantitative polymerase chain reaction were used to detect FOXO3 expression in pancreatic cancer cells and human pancreatic stellate cells. PANC-1 and MIAPaCa-2 pancreatic cancer cells with low FOXO3 expression were selected to transfect FOXO3 overexpression plasmid and negative control plasmid, respectively. The motility and proliferation ability of pancreatic cancer cells were detected by colony formation assay, cell scratch assay, Transwell assay and flow cytometry.Results:In the LinkedOmics database, the relative expression of FOXO3 protein in the cancer tissues of 64 patients with pancreatic cancer was significantly lower than that in the adjacent tissues ( t=8.36, P<0.001). The number of clones in PANC-1 cell line was (30.0±6.6) after overexpressed FOXO3, which was lower than that in negative control cells (92.7±6.7), and the difference was statistically significant ( t=11.54, P<0.001). After overexpressed FOXO3 in PANC-1 and MIAPaCa-2 cell lines, the scratch repair rate was significantly decreased compared with the control group. In Transwell experiment, the number of cells in FOXO3 overexpressed group in PANC-1 cell lines was (21.0±6.6), which was lower than that of negative control groups (55.7±8.5), and the difference was statistically significant ( t=5.59, P=0.005). The results of MIAPaCa-2 cell line were consistent with that of PANC-1 cell line. After overexpressing of FOXO3 in PANC-1 and MIAPaCa-2 cell lines, the proportion of cells in the G0/G1 phase decreased, while the proportion in the S phase increased. Conclusion:The expression of FOXO3 was decreased in pancreatic cancer. Overexpression of FOXO3 could significantly inhibit the proliferation, migration and invasion of pancreatic cancer cells and induce cell cycle arrest, which is a potential target for the treatment of pancreatic cancer.

The influence of Jianpi Qinghua formula on skeletal muscle protein accompanied by insulin resistance by increasing FoxO1 phosphorylation / 国际中医中药杂志

Objective:Based on the degradation of skeletal muscle protein MuRF mediated by pFoxO1 when insulin resistance occurs, this paper explores the content change of skeletal muscle protein and the effect of Jianpi Qinghua formula when insulin resistance occurs.Methods:C57 mice were fed with high-fat food and made as the model of obesity accompanied by insulin resistance. Then they were divided into model group, Jianpi Qinghua formula group and metformin group according to random number table method with 10 mice in each group. Jianpi Qinghua formula group was orally administered with water decoction 20.961 g/kg, and the metformin group was orally administered with metformin suspension 18.498 g/kg, once a day for 12 consecutive weeks. Intraperitoneal Glucose Tolerance Tests (IPGTT) was used after the model was established and intervened respectively. The relative protein content of pFoxO1, FoxO1, MuRF, MyoD and myosin were detected by Western blot method, and the localization of MyoD and myosin was detected by immuno-histochemistry.Results:Compared with the model group, the blood glucose of IPGTT at 0 min, 60 min and 120 min of both Jianpi Qinghua formula group and Metformin group decreased ( P<0.05). Compared with model group, the ratio of pFoxO1/FoxO1 protein expression level (0.27±0.07, 0.24±0.14 vs. 0.05±0.03) of both Jianpi Qinghua formula group and Metformin group increased ( P<0.05), and the relative expression of MuRF protein (1.22±0.42, 1.15±0.32 vs. 3.21±0.35) of both Jianpi Qinghua formula group and Metformin group decreased ( P<0.05). The relative protein expression of MyoD (1.42±0.45 vs. 0.40±0.11) and myosin (0.80±0.11 vs. 0.51±0.08) relative protein expression of Jianpi Qinghua formula group was significantly higher than that of model group ( P<0.05). Immunohistochemical staining showed that MyoD (5.06±1.72 vs.2.28±0.83) and myosin (60.28±7.47 vs. 39.77±3.34) of Jianpi Qinghua formula group significantly increased compared with model group ( P<0.05). Conclusion:Jianpi Qinghua formula could effectively increase the content of skeletal muscle protein, enhancing the phosphorylation of FoxO1 in skeletal muscle and the inhibition of MuRF degradation pathway.

Research progress of FOXO1 in oral squamous cell carcinoma / 国际肿瘤学杂志

Forkhead box protein O1 (FOXO1) has been extensively studied as a tumor suppressor. In oral squamous cell carcinoma, studies have demonstrated that FOXO1 can inhibit tumor cell oxidative stress, stemness and epithelial-mesenchymal transition, and promote tumor cell autophagy and apoptosis. FOXO1 may serve as a potential target for the treatment of oral squamous cell carcinoma.

Mechanism of FOXA2 in colorectal cancer and its application in diagnosis and treatment / 国际肿瘤学杂志

Forkhead box protein A2 (FOXA2) , characterized by its unique DNA binding domain, plays a key role in transcriptional regulation. FOXA2 transcription factor is highly expressed in colorectal cancer, and binds with corresponding targeted genes to regulate tumor growth and inflammatory response, thus playing the role of oncogenes. In-depth understanding of the role and function of human FOXA2 transcription factor in colorectal cancer will contribute to further development of FOXA2 as a potential therapeutic target and diagnostic and prognostic marker of colorectal cancer.

Effects of astragalin on the cell proliferation and cell cycle of prostate cancer cells through up-regulating miRNA-513 expression / 肿瘤研究与临床

Objective:To investigate the effects of astragalin on the cell proliferation and cell cycle of prostate cancer cell line C4-2B through up-regulating the expression of miRNA-513 (miR-513).Methods:Prostate cancer cell line C4-2B cells were taken and treated with 125 μg/L of astragalin for 48 h (astragalin group), and untreated C4-2B cells were set as the control group. The methyl thiazolyl tetrazolium (MTT) method was used to detect the proliferation ability of C4-2B cells in the two groups, and cell cycle was detected by using flow cytometry. The miRNAMap prediction software was used to predict that the targeted gene of miR-513 was the forkhead box protein R2 (FOXR2), and the dual luciferase gene reporter assay was used to verify it. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of miR-513 and FOXR2 mRNA in the two groups of cells. Western blotting was used to detect the expressions of FOXR2, cyclin-dependent kinase 7 (CDK7), β-actin and cyclin H in the two groups of C4-2B cells.Results:Compared with the control group, the proliferation activity of C4-2B cells in the astragalin group was decreased from day 2 to day 5 (all P < 0.05). The proportions of S-phase cells in the control group and the astragalin group were (48.1±3.2)% and (36.0±2.1)%, respectively. The proportion of S-phase cells in the astragalin group was decreased ( t = 3.12, P = 0.021); the proportions of G 2-phase cells were (24.9±3.3)% and (11.8±2.4)%, respectively. The proportion of G 2-phase cells in the astragalin group was decreased ( t = 3.18, P = 0.019). The relative expression levels of miR-513 in C4-2B cells of the control group and the astragalin group were 1.01±0.22 and 6.55±0.61, respectively. The relative expression levels of miR-513 in C4-2B cells in the astragalin group was increased ( t = 7.70, P < 0.01). The dual luciferase reporter gene assay verified that FOXR2 was the targeted gene of miR-513. The relative expression level of FOXR2 mRNA in C4-2B cells of the control group and the astragalin group was 1.04±0.14 and 0.19±0.06, respectively, and the difference was statistically significant ( t = 5.53, P = 0.002), suggesting that after astragalin promoted the expression of miR-513, the FOXR2 mRNA expression was decreased. The relative expression levels of FOXR2, CDK7 and cyclin H protein in C4-2B cells in the astragalin group were all decreased compared with those in the control group. Conclusions:Astragalin inhibits the proliferation of prostate cancer C4-2B cells and induces cell cycle arrest by up-regulating the expression of miR-513.

Efficacy of anti-CD 20 monoclonal antibody in combination with Gemcitabine-based chemotherapy regimen in children with recurrent diffuse large B-cell lymphoma / 中华实用儿科临床杂志

Objective:To investigate the efficacy of anti-CD 20 monoclonal antibody in combination with Gemcitabine-based chemotherapy regimen in children with recurrent diffuse large B-cell lymphoma (DLBCL). Methods:The clinical data of 62 children with DLBCL admitted to the Department of Pediatrics, Yantai Mountain Hospital of Yantai City and the Department of Pediatrics, Muping District Traditional Chinese Medicine Hospital in Yantai City from January 2010 to January 2018 were analyzed retrospectively.Different treatment options were selected according to the children′s stage and the presence of risk factors such as huge tumors.Among them, 32 cases in the control group were treated with the Gemcitabine-based treatment plan (Gemcitabine+ Cisplatin+ Dexamethasone treatment). Thirty patients in the study group were treated with anti-CD 20 monoclonal antibody on the basis of the control group for a total of 4 cycles (21 d per cycle). After 4 cycles of treatment, the clinical efficacy, the positive expression of forkhead box protein P1 (FOXP1) and B-cell lymphoma factor-6 (Bcl-6) before and after treatment, the occurrence of adverse reactions and survival[3-year progression-free survival (PFS), 3-year overall survival (OS)] were evaluated.The measurement data that meets the normal distribution is expressed that the t test is used, and the counting data is represented by (%), and the χ2 test is used.Level data is compared with ranking and inspection. Results:All patients were followed up to March 2021.The total response rate (RR) and disease control rate (DCR) of the study group were 93.33% (28/30 cases) and 96.67% (29/30 cases), respectively.The RR and DCR of the control group were 68.75% (22/32 cases) and 81.25% (26/32 cases), respectively.The RR of the study group was higher than that of the control group ( χ2=5.995, P<0.05), and there was no statistical difference in DCR between the two groups ( χ2=3.674, P>0.05). After treatment, the positive expression rate of FOXP1 in the study group and the control group was lower than before treatment[23.33% (7/30 cases) vs. 76.67% (23/30 cases); 50.00% (16/32 cases) vs. 75.00% (24/32 cases), χ2=17.067, 4.267, all P<0.05], and the positive expression rate of the study group was lower than that of the control group ( χ2=4.179, P<0.05). After treatment, the positive expression rate of Bcl-6 in the study group and the control group was higher than before treatment[86.67% (26/30 cases) vs. 26.67% (8/30 cases); 62.50% (20/32 cases) vs. 31.25% (10/32 cases), χ2=21.991, 6.275, all P<0.05], and the positive expression rate of the study group was higher than that of the control group ( χ2=4.723, P<0.05). There was no significant difference between the study group and the control group in the level of gastrointestinal reactions, elevated transa-minase, and decreased white blood cell ( Z=-1.074, -1.078, -0.834, all P>0.05). There was a difference between the 3-year PFS survival curve and the 3-year OS survival curve between the two groups ( χ2=3.997, 4.723, all P<0.05). Conclusions:Anti-CD 20 monoclonal antibody combined with Gemcitabine-based chemotherapy is effective for children with DLBCL, and will not significantly increase the adverse reactions in children.

FOXP3 variants are independently associated with transforming growth factor B1 plasma levels in female patients with inflammatory bowel disease

Abstract Objective: The aim of this study was to evaluate the association of -924 G>A (rs2232365) and -3279 C>A (rs3761548) FOXP3 variants with IBD susceptibility, clinical and endoscopic activity, and IL-10 and TGF-β1 plasma levels. Method: The study included 110 IBD female patients, 60 with Ulcerative Colitis (UC) and 50 with Crohn's Disease (CD), and 154 female controls. FOXP3 variants were determined with Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP). Plasma levels of IL-10 and TGF-β1 were determined using immuno-fluorimetric assay. Results: AA genotype of rs2232365 and rs3761548 was associated with CD (OR = 3.147, 95% CI 1.015-9.758, p = 0.047) and UC (OR = 3.221, 95% CI 1.050-9.876, p = 0.041) susceptibility, respectively. However, were not associated with TGF-β1 and IL-10 levels, and endoscopic/clinical activity disease. GAGA haplotype was associated with IBD (OR = 4.003, 95% CI 1.100-14.56, p = 0.035) and UC susceptibility (OR = 6.107, 95% CI 1.609-23.18, p = 0.008). In addition, IBD patients with the GAGA haplotype had lower TGF-β1 levels (p = 0.041). Moreover, G/C haplotype (dominant model) had a protective effect of 60% in CD susceptibility and lower Endoscopic Severity Index. Conclusions: These results suggest that FOXP3 variants could exert a role in the Treg, which could be one of the factors involved in the susceptibility and pathogenesis of IBD.

Role of Foxo6 in insulin resistance of vascular endothelial cells induced by high glucose and high fat and its interaction with NF-κB pathway / 中国药理学通报

Aim To investigate the expression of Foxos in human umbilical vein endothelial cells(HUVECs)with insulin resistance(IR)induced by high glucose and high fat(HG/HF)stress and its significance.Methods First, the IR model of endothelial cells was established by HG /HF stress.The differential expression of Foxos gene in normal(Ctrl )group and HG /HF group was observed, and the subtypes with the most significant changes in Foxos were screened out, such as Foxo6.Next, Foxo6 was silenced to observe its role in endothelial cell with IR.Finally, whether the mechanism of Foxo6-mediated IR was related to the interaction of NF-κB signaling was investigated.Results The expression increase of Foxo6 was the most significant among Foxos under the IR condition induced by HG/HF.Using a small RNA interference and plasmid transfection technique, we found that the silence effect of the siRNA3 fragments targeting Foxo6 was the most significant among the siRNAs.Moreover, the further study showed that silencing the Foxo6 gene could significantly reverse the endothelial IR induced by HG/HF, and the mechanism of the reversal effect was related to the interaction between the Foxo6 and NF-κB signal.Conclusions Foxo6 mediates the endothelial cell IR induced by the HG /HF stress.The underlying mechanism is that Foxo6 can interact with NF-κBp65 and activate NF-κB signaling pathway.Silencing Foxo6 can improve the IR of vascular endothelial cells induced by HG /HF stress.

Correlation of macrophage-related cytokines and silent information regulator 1 and forkhead box protein O3 levels in peripheral blood mononuclear cells in patients with active pulmonary tuberculosis / 中国热带医学

@#Abstract: Objective　To explore the correlation between the levels of silent information regulator 1 (SIRT1) and forkhead box protein O3 (FOXO3) in peripheral blood mononuclear cells of patients with active pulmonary tuberculosis (APTB) and macrophage-related cytokines-inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1). Methods　A total of 64 APTB patients who were treated in Yubei Hospital, the First Affiliated Hospital of Chongqing Medical University from January 2020 to December 2021 were gathered as the APTB group, 59 people with latent tuberculosis infection (LTBI) were gathered as the LTBI group, and 62 healthy people were gathered as the control group. Quantitative real-time PCR (qPCR) method was performed to measure the levels of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells. The enzyme-linked immunosorbent assay (ELISA) was performed to measure serum iNOS and Arg-1 levels; ROC curve was used to analyze the value of SIRT1 mRNA and FOXO3 mRNA levels in the differential diagnosis of LTBI and APTB; Pearson correlation was performed to analyze the correlation of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells of APTB patients with serum iNOS and Arg-1 levels. Results　The levels of SIRT1 mRNA, FOXO3 mRNA and serum iNOS in peripheral blood mononuclear cells decreased in control group, LTBI group and APTB group, and the level of serum Arg-1 increased in turn (P<0.05). The AUCs of SIRT1 mRNA and FOXO3 mRNA in differential diagnosis of LTBI and APTB were 0.876 and 0.887, respectively, the sensitivity was 71.2% and 76.3%, and the specificity was 96.9% and 90.6% respectively. The levels of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells of APTB patients were positively correlated (r=0.500, P<0.05), and they were positively correlated with serum iNOS and negatively correlated with serum Arg-1 (P<0.05). The SIRT1 mRNA, FOXO3 mRNA and serum iNOS in peripheral blood mononuclear cells of APTB patients after 6 months of treatment were higher than those before treatment, and serum Arg-1 was lower than before treatment (P<0.05). Conclusions　The levels of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells of APTB patients are low, and they are positively correlated with macrophage-related cytokine iNOS and negatively correlated with Arg-1.

Atractylenolide III alleviates sepsis-mediated lung injury via inhibition of FoxO1 and VNN1 protein

ABSTRACT Purpose: To evaluate the influence of atractylenolide (Atr) III on sepsis-induced lung damage. Methods: We constructed a mouse sepsis model through cecal ligation and puncture. These mice were allocated to the normal, sepsis, sepsis + Atr III-L (2 mg/kg), as well as Atr III-H (8 mg/kg) group. Lung injury and pulmonary fibrosis were accessed via hematoxylin-eosin (HE) and Masson's staining. We used terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and flow cytometry for detecting sepsis-induced lung cell apoptosis. The contents of the inflammatory cytokines in lung tissue were measured via enzyme-linked immunosorbent assay (ELISA). Results: Atr III-H did not only reduce sepsis-induced lung injury and apoptosis level, but also curbed the secretion of inflammatory factors. Atr III-H substantially ameliorated lung function and raised Bcl-2 expression. Atr III-H eased the pulmonary fibrosis damage and Bax, caspase-3, Vanin-1 (VNN1), as well as Forkhead Box Protein O1 (FoxO1) expression. Conclusions: Atr III alleviates sepsis-mediated lung injury via inhibition of FoxO1 and VNN1 protein.

Effect and potential molecular mechanism of FOXA2 on cell proliferation, migration and invasion of hepatocellular carcinoma / 中华肝胆外科杂志

Objective:To investigate the effect of forkhead box protein A2(FOXA2) on cell proliferation, migration and invasion of hepatocellular carcinoma and the potential molecular mechanism.Methods:From January 2019 to December 2020, 10 cases of hepatocellular carcinoma patients from Zhongnan Hospital of Wuhan University were collected for study, including 7 males and 3 females, with an average age of 53 years. FOXA2 expression was detected in human liver cancer cell line, and the highest expression of FOXA2 was found in HepG2 cells transfected with FOXA2 overexpression plasmid. Immunohistochemistry and qRT-PCR were used to detect the expression of FOXA2. Western blot was used to detect the expression of FOXA2, hypoxia-inducible factor-1 α (HIF-1α), vascular endothelial growth factor A (VEGFA), B-cell lymphofactor-2 (Bcl-2), matrix metalloproteinase (MMP) 7, and glucose transporter (GLUT) 1. EdU assay was used to study cell proliferation, and Transwell chamber assay was used to study cell migration and invasion.Results:The relative expression of FOXA2 in liver cancer tissues were lower than those in adjacent tissues both at mRNA and protein levels, with statistical significance (both P<0.05). FOXA2 overexpression group showed lower cell proliferation rate (30.0±3.2)%, migration rate (10.6±1.1), and invasion rate (12.8±0.8) comparing with negative control group (67.0±3.6)%, (81.0±5.4), (74.8±4.5). The difference was statistically significant (all P<0.05). Expression of HIF-1α and its downstream targets VEGFA, MMP7, GLUT1 and Bcl-2 was decreased after over-expression of FOXA2 in HepG2 cells. Conclusion:FOXA2 inhibits proliferation, migration, and invasion in hepatocellular carcinoma by regulating HIF-1α signaling pathway, suggesting that FOXA2 is a potential target for the treatment of hepatocellular carcinoma.

Proteolysis Pathways via FoxO1 Reduce the Diameter of Myotube under Hypoxia Exposure / 中国生物化学与分子生物学报

The high altitude/hypoxic environment induced skeletal muscle atrophy is considered to be the interaction of multi-system and multi-organ, but the direct mechanism of hypoxia on muscle cells in this process is not clear. This study intended to investigate the effects of hypoxia exposure on proteins in ubiquitin and autophagy pathways, and explored the possible mechanism of hypoxia induced change of myotube diameter. The expression of myosin, hypoxia inducible factor-1 α (HIF-1α), forkhead box protein O1 (FoxO1), and ubiquitin protease pathway (MuRF1 and Atrogin1) and autophagy lysosomal pathway (p62, Beclin1, LC3) related proteins were detected by Western blot; The integrated optical density (IOD) of Myosin and LC3 was detected by IF. The results showed that the diameters of myotube at 6 h and 12 h were significantly reduced, and the expression of myosin was significantly reduced at 6 h after hypoxia exposure (P<0. 05); the protein levels of HIF-1α and FoxO1 were significantly increased at 6 h (P<0. 05); The expression of MuRF1 in each time points of hypoxia was significantly higher than 0 h (P<0. 05), but no difference of Atrogin1 expression was detected; Compared with 0 h, the expression of p62 was reduced significantly in response to hypoxia. The protein expression of Beclin1 and the IOD of LC3 was increased significantly at 6 h, and the LC3Ⅱ/Ⅰ ratio was significantly higher at 6 h, but significantly lower at 12 h and 24 h (P<0. 05).The results above indicated that the reduction of the myotube diameter of L6 skeletal muscle cells was induced by hypoxia exposure (1% O

Establishment of human embryonic stem cell line with forkhead box G1 gene knockout by CRISPR/Cas9 / 第二军医大学学报

Objective To construct human embryonic stem cell (hESC) line with forkhead box G1 (FOXG1) gene knockout by clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing technology, and to investigate the role of FOXG1 gene in the early neural induction of hESCs. Methods Two guide RNAs (gRNAs) were transfected to induce FOXG1 gene large fragment knockout in hESCs by CRISPR/Cas9 gene editing technology. FOXG1 gene knockout hESCs were confirmed by monoclonal screening, sequencing and Western blotting analysis. The expression of the key markers including paired box 6 (PAX6), sex-determining region Y-box 2 (SOX2) and orthodenticle homeobox 2 (OTX2) was detected by immunofluorescence staining and qRT-PCR in the early process of neural induction before and after FOXG1 gene knockout. Results hESCs with FOXG1 gene large fragment knockout were successfully obtained by CRISPR/Cas9 gene editing technology. The results of immunofluorescence staining and qRTPCR suggested that FOXG1 deletion did not significantly influence the expression of PAX6, SOX2 and OTX2 during neural induction. Conclusion FOXG1 gene large fragment knockout in hESCs can be quickly induced by a pair of gRNAs cotransfection. FOXG1 deletion has no significant impacts on neural induction of hESCs.

Value of human mitochondrial transcription termination factor 3 and forkhead box protein 3 in predicting the prognosis of non-small cell lung cancer / 肿瘤研究与临床

Objective@#To explore the prognostic value of human mitochondrial transcription termination factor 3 (hMTERF3) and forkhead box protein 3 (Foxp3) in non-small cell lung cancer (NSCLC).@*Methods@#The clinical data of 88 patients with NSCLC who were admitted to the Third Medical Center of PLA General Hospital from March 2017 to March 2018 were retrospectively analyzed. All patients were diagnosed by pathological puncture. The patients were followed-up by telephone for 12 months, and according to the prognosis, the patients were divided into good prognosis group and poor prognosis group. The pathological tissues were taken from all patients, and the expressions of hMTERF3 and Foxp3 proteins were detected by immunohistochemistry. The expressions of hMTERF3 and Foxp3 in the good prognosis group and the poor prognosis group were compared. Logistic regression model was used to analyze the risk factors of poor prognosis in patients with NSCLC.@*Results@#Of 88 patients, 61 patients (69.3%) had good prognosis and 27 patients (30.7%) had poor prognosis. The positive expression rate of hMTERF3 in the good prognosis group was 57.4% (35/61), which was significantly lower than that in the poor prognosis group (81.5%, 22/27) (χ 2= 4.766, P= 0.029). The positive expression rate of Foxp3 in the good prognosis group was 55.7% (34/61), which was significantly lower than that in the poor prognosis group (85.2%, 23/27) (χ 2= 7.113, P= 0.008). The proportions of patients with medium and high differentiation or stage Ⅰ- Ⅱ in the good prognosis group were 82.0% (50/61) and 68.8% (42/61), respectively, which were significantly higher than those in the poor prognosis group [48.15% (13/27) and 25.93% (7/27)] (both P < 0.05). Logistic regression analysis showed that the poor differentiation, stage Ⅲ-Ⅳ, hMTERF3-positive and Foxp3-positive were the risk factors for poor prognosis in NSCLC patients (all P < 0.05).@*Conclusions@#The positive expression rates of hMTERF3 and Foxp3 in patients with good prognosis are lower. The hMTERF3-positive and Foxp3-positive are risk factors for poor prognosis in NSCLC patients.

Effect of lncRNA HULC knockdown on rat secreting pituitary adenoma GH3 cells

Pituitary adenoma is one of the most common tumors in the neuroendocrine system. This study investigated the effects of long non-coding RNAs (lncRNAs) highly up-regulated in liver cancer (HULC) on rat secreting pituitary adenoma GH3 cell viability, migration, invasion, apoptosis, and hormone secretion, as well as the underlying potential mechanisms. Cell transfection and qRT-PCR were used to change and measure the expression levels of HULC, miR-130b, and FOXM1. Cell viability, migration, invasion, and apoptosis were assessed using trypan blue staining assay, MTT assay, two-chamber transwell assay, Guava Nexin assay, and western blotting. The concentrations of prolactin (PRL) and growth hormone (GH) in culture supernatant of GH3 cells were assessed using ELISA. The targeting relationship between miR-130b and FOXM1 was verified using dual luciferase activity. Finally, the expression levels of key factors involved in PI3K/AKT/mTOR and JAK1/STAT3 pathways were evaluated using western blotting. We found that HULC was highly expressed in GH3 cells. Overexpression of HULC promoted GH3 cell viability, migration, invasion, PRL and GH secretion, as well as activated PI3K/AKT/mTOR and JAK1/STAT3 pathways. Knockdown of HULC had opposite effects and induced cell apoptosis. HULC negatively regulated the expression of miR-130b, and miR-130b participated in the effects of HULC on GH3 cells. FOXM1 was a target gene of miR-130b, which was involved in the regulation of GH3 cell viability, migration, invasion, and apoptosis, as well as PI3K/AKT/mTOR and JAK1/STAT3 pathways. In conclusion, HULC tumor-promoting roles in secreting pituitary adenoma might be via down-regulating miR-130b, up-regulating FOXM1, and activating PI3K/AKT/mTOR and JAK1/STAT3 pathways.