FOXM1 regulates the proliferation, apoptosis and inflammatory response of keratinocytes through the NF-κB signaling pathway

2021 ◽  
pp. 096032712098422
Author(s):  
Mi Zhou ◽  
Jing Shi ◽  
Shaobo Lan ◽  
Xianjun Gong
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Caspase 3 ◽  
Normal Skin ◽  
Psoriatic Skin ◽  
Hacat Cells ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Tnf Α

Psoriasis is a common immune-mediated and genetic skin disease. Forkhead box M1 (FOXM1) is a member of FOX family that has been found to modulate skin disorders. However, its role in psoriasis remains unknown. Thus, we aimed to investigate the effect of FOXM1 on keratinocytes in response to tumor necrosis factor-α (TNF-α). The expression levels of FOXM1 in psoriasis tissues and normal skin tissues were examined using qRT-PCR and western blot. HaCaT cells were stimulated by TNF-α to mimic psoriasis in vitro. MTT assay was performed to assess cell proliferation. The caspase-3 activity and expression levels of bcl-2 and bax were determined to indicate cell apoptosis. The mRNA and secretion levels of IL-6, IL-23 and TGF-β were determined by qRT-PCR and ELISA, respectively. The NF-κB activation was assessed using western blot analysis. Our results demonstrated that FOXM1 was highly upregulated in psoriatic skin tissues and TNF-α-stimulated HaCaT cells. Knockdown of FOXM1 repressed cell proliferation of TNF-α-stimulated HaCaT cells. Knockdown of FOXM1 caused significant increases in caspase-3 activity, bax expression and decrease in bcl-2 expression in TNF-α-stimulated HaCaT cells. Moreover, FOXM1 knockdown also suppressed the TNF-α-induced production of IL-6, IL-23, and TGF-β in HaCaT cells. However, FOXM1 overexpression showed the opposite effect. Furthermore, the TNF-α-induced NF-κB activation was prevented by FOXM1 knockdown. Additionally, inhibition of NF-κB reversed the effects of FOXM1 on HaCaT cells. Taken together, these findings indicated that FOXM1 regulated cell proliferation, apoptosis and inflammation in TNF-α-induced HaCaT cells. The effects of FOXM1 were mediated by NF-κB pathway.

scholarly journals Anti-breast cancer effect and mechanism of ethyl acetate extract of persimmon leaves

2020 ◽  
Author(s):  
Fang Zhuge ◽  
Xiusha Wei ◽  
Yani Wu ◽  
Guining Wei ◽  
Ming Chen ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Ethyl Acetate ◽  
Caspase 3 ◽  
Ethyl Acetate Extract ◽  
Tumor Inhibition ◽  
Expression Levels ◽  
Tnf Α ◽  
Tumor Inhibition Rate

Abstract Aim: The anti-breast cancer effect and mechanism of ethyl acetate extract of persimmon leaves (PLE) were determined. Methods: Persimmon leaves were extracted by reflux at 80°C with 80% ethanol as the solvent. The total extracts of persimmon leaves were extracted with ethyl acetate, and the yield was calculated by weighing. The mouse breast cancer cell line 4T1 was cultured in vitro, and different concentrations of PLE were added. At the same time, the effects of PLE at different concentrations (25, 50, 100 µg/ml) on cell apoptosis ability were detected by Acridine Orange and Ethidium Bromide (AOEB) and flow cytometry experiments. In addition, real-time quantitative PCR (real-time PCR, RT-PCR) was used to test the expression of the Bax, Bcl-2, ERK1/2, MEK1/2 and RAF genes. In vivo tumor-bearing mouse model: A breast cancer transplant tumor model was established with BALB/c mice. The doses of PLE were 30, 60 and 120 mg/kg body weight/d, and the dose of CTX was 20 mg/kg body weight/d. The tumor inhibition rate and the effects of PLE on immune organs in tumor-bearing mice with 4T1 breast cancer were determined. The expression levels of IL-6, TNF-α, TGF-β and VEGFA in the serum of mice were detected by ELISA. The expression of the Bax, Bcl2, ERK1/2, MEK1/2 and RAF genes was determined by RT-PCR. The protein expression levels of Bax, Bcl-2, Caspase-3, p-MEK, p-JNK and p-P38 in tumor tissues were detected by immunohistochemistry. In addition, the protein expression levels of MAPK pathway components were assessed through Western blotting.Results: A total of 119.34 g ethyl acetate extract was obtained from 3 kg persimmon leaves with a yield of 3.98%. In vitro: MTT results indicated a strong antiproliferative effect of PLE on breast cancer cell lines. AOEB and flow cytometry assays showed that PLE promoted the apoptosis of breast cancer cells. PCR results showed that PLE could inhibit Bcl-2, promote Bax expression, and downregulate ERK1/2, MEK1/2, and RAF gene expression. In vivo: PLE had a significant inhibitory effect on breast cancer, and the tumor inhibition rates were 11.65%, 33.71% and 47.24% from low dose to high dose, respectively, showing a concentration dependence. The tumor inhibition rate of CTX was 57.74%. Meanwhile, PLE can increase the spleen and thymus index of 4T1 mice and decrease the liver index of 4T1 mice. Compared with the model group, PLE significantly reduced the expression levels of IL-6, TNF-α, TGF-β and VEGFA in the serum of mice. PCR results showed that PLE could inhibit Bcl-2, promote Bax expression, and downregulate ERK1/2, MEK1/2, and RAF gene expression. Immunohistochemical results showed that the PLE group and CTX group significantly promoted the expression of Bax and Caspase-3 proteins and downregulated the expression of Bcl-2, p-MEK, p-JNK and p-P38 proteins. WB results showed that PLE regulated the expression of proteins in the MAPK pathway.Conclusion: PLE enhances immunity, inhibits angiogenesis, inhibits 4T1 cell proliferation and induces apoptosis. Its apoptosis mechanism is related to the regulation of Bax/Bcl-2/Caspase-3 protein and the phosphorylation of regulatory proteins related to the MAPK signaling pathway.

scholarly journals CircRNA_100290 Promotes GC Cell Proliferation And Invasion Via miR-29b-3p/ITGA11 Axis And Is Regulated By EIF4A3

2021 ◽  
Author(s):  
Gang Wang ◽  
Dan Sun ◽  
Wenhui Li ◽  
Yan Xin
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Reporter Assay ◽  
Qrt Pcr ◽  
Node Metastasis ◽  
Dual Luciferase Reporter Assay

Abstract Background: Circular RNA (circRNA) has been reported as an important regulator in the development and progression of various carcinomas. However, the role of circRNA_100290 in gastric cancer (GC) is still unclear. This study aimed to investigate the role of circRNA_100290 in GC invasion and metastasis and its possible mechanism.Methods: The expression of circRNA_100290 in GC cells and tissues were examined using quantitative real-time polymerase chain reaction (qRT-PCR). The role of circRNA_100290 in cell proliferation, migration, and invasion was evaluated on AGS and HGC-27 cell lines in vitro. Bioinformatics tools, dual-luciferase reporter assay, Western blot assay and qRT-PCR were used to explore the downstream pathways of circRNA_100290. The mechanism underlying the regulation of the expression of circRNA_100290 was explored using RNA immunoprecipitation, qRT-PCR, and Western blot assays.Results: The expression of circRNA_100290 was found significantly upregulated in GC cells and 102 GC tissues, high expression of circRNA_100290 in GC was closely related to Borrmann’s types, lymph node metastasis and tumor-node-metastasis staging. In vitro, knockdown of circRNA_100290 in AGS and HGC-27 cells significantly inhibited cell proliferation, migration, and invasion. Mechanistically, dual-luciferase reporter assay confirmed a direct binding between circRNA_100290 and miR-29b-3p, which targets ITGA11, an oncogene which is closely related to epithelial–mesenchymal transition (EMT). In addition, EIF4A3, one of RNA binding proteins (RBPs), could inhibit the formation of circRNA_100290 via enriching flanking sites of circRNA_100290. Low expression of EIF4A3 in GC was related to a worse prognosis.Conclusions: Elevated circRNA_100290 in GC promotes cell proliferation, invasion and EMT via miR-29b-3p/ITGA11 axi and might be regulated by EIF4A3. CircRNA_100290 might be a promising biomarker and target for GC therapy.

scholarly journals CircRNA_100290 promotes GC cell proliferation and invasion via the miR-29b-3p/ITGA11 axis and is regulated by EIF4A3

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Gang Wang ◽  
Dan Sun ◽  
Wenhui Li ◽  
Yan Xin
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Rna Binding ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Mesenchymal Transition ◽  
Qrt Pcr ◽  
Node Metastasis

Abstract Background Circular RNAs (circRNAs) have been reported to be important regulators of the development and progression of various carcinomas. However, the role of circRNA_100290 in gastric cancer (GC) is still unclear. This study aimed to investigate the role of circRNA_100290 in GC invasion and metastasis and the possible underlying mechanism. Methods The expression of circRNA_100290 in GC cells and tissues was examined using quantitative real-time polymerase chain reaction (qRT-PCR). The role of circRNA_100290 in cell proliferation, migration, and invasion was evaluated in the AGS and HGC-27 cell lines in vitro. Bioinformatics tools, dual-luciferase reporter assays, Western blot assays and qRT-PCR were used to explore the pathways downstream of circRNA_100290. The mechanism underlying the regulation of circRNA_100290 expression was explored using RNA immunoprecipitation, qRT-PCR, and Western blot assays. Results The expression of circRNA_100290 was significantly upregulated in GC cells and 102 GC tissues, and high circRNA_100290 expression in GC was closely related to Borrmann’s type, lymph node metastasis and tumour-node-metastasis stage. In vitro, knockdown of circRNA_100290 in AGS and HGC-27 cells significantly inhibited cell proliferation, migration, and invasion. Mechanistically, a dual-luciferase reporter assay confirmed the direct interaction between circRNA_100290 and miR-29b-3p, which targets ITGA11, an oncogene that is closely related to epithelial–mesenchymal transition (EMT). In addition, EIF4A3, an RNA-binding protein (RBP), could inhibit the formation of circRNA_100290 by binding to the flanking sites of circRNA_100290. Low EIF4A3 expression in GC was related to a poor prognosis. Conclusions Elevated circRNA_100290 expression in GC promotes cell proliferation, invasion and EMT via the miR-29b-3p/ITGA11 axis and might be regulated by EIF4A3. CircRNA_100290 might be a promising biomarker and target for GC therapy. Graphical abstract

EphrinB2 promotes the human aortic smooth muscle cell growth and migration via mediating F-actin remodeling

Vascular ◽  
2021 ◽  
pp. 170853812110521
Author(s):  
Fan Zhu ◽  
Jia Chen ◽  
Mingyao Luo ◽  
Dongting Yao ◽  
Xiaobo Hu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Smooth Muscle ◽  
Aortic Dissection ◽  
Western Blot ◽  
Prognostic Biomarker ◽  
Migration And Invasion ◽  
Aortic Smooth Muscle ◽  
Qrt Pcr

Objectives To evaluate the potential effect of EphrinB2 in human thoracic aortic dissection (TAD) and to illustrate the mechanisms governing the role of EphrinB2 in the growth of human aortic smooth muscle cells (HASMC). Methods In the study, EphrinB2 expression was investigated by qRT-PCR and immunohistochemistry in 12 pairs of TAD and adjacent human tissues. HASMCs were used for in vitro experiments. Next, EphrinB2 overexpression and depletion in HASMCs were established by EphrinB2-overexpressing vectors and small interfering RNA, respectively. The transfection efficiency was evaluated by qRT-PCR and Western blot. The effects of overexpression and depletion of EphrinB2 on cell proliferation, migration, and invasion were tested in vitro. Cell Counting Kit-8, flow cytometry and transwell migration/invasion, and wound healing assay were used to explore the function of EphrinB2 on HASMC cell lines. The relationship between EphrinB2 and F-actin was assessed by Western blot, immunofluorescence, and Co-IP. Results We found that EphrinB2 was a prognostic biomarker of TAD patients. Moreover, EphrinB2 expression negatively correlated to aortic dissection tissues, and disease incidence of males, suggesting that EphrinB2 might act as a TAD suppressor by promoting proliferation or decreasing apoptosis in HASMC. Next, over-expression of EphrinB2 in HASMC lines drove cell proliferation, migration, and invasion, and inhibited apoptosis while knockdown EphrinB2 showed the opposite phenomenon, respectively. Furthermore, the level of F-actin in mRNA, protein, and distribution in HASMC cell lines highly matched with the expression of EphrinB2, which indicated that EphrinB2 could mediate the HASMC cytoskeleton via inducing F-actin. Conclusions In conclusion, our results first provided the pivotal role of EphrinB2 in HASMC proliferation initiated by mediating F-actin and demonstrated a prognostic biomarker and the potential targets for therapy to prevent thoracic aortic dissection.

scholarly journals EIF4A3-Mediated CircRNA_100290 Promotes GC Cell Proliferation, Invasion and EMT via miR-29b-3p/ITGA11 Axis

2020 ◽  
Author(s):  
Gang Wang ◽  
Dan Sun ◽  
Wenhui Li ◽  
Yan Xin
Keyword(s):  
Cell Proliferation ◽  
Western Blot ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Reporter Assay ◽  
Qrt Pcr ◽  
Node Metastasis ◽  
Dual Luciferase Reporter Assay

Abstract Background Circular RNA (circRNA) has been reported as an important regulator in the development and progression of various carcinomas. However, the role of circRNA_100290 in gastric cancer (GC) is still unclear. This study aimed to investigate the role of circRNA_100290 in GC invasion and metastasis and its possible mechanism.Methods The expression of circRNA_100290 in GC cells and tissues were examined using quantitative real-time polymerase chain reaction (qRT-PCR). The role of circRNA_100290 in cell proliferation, migration, and invasion was evaluated on AGS and HGC-27 cell lines in vitro. Bioinformatics tools, dual-luciferase reporter assay, Western blot assay and qRT-PCR were used to explore the downstream pathways of circRNA_100290. The mechanism underlying the regulation of the expression of circRNA_100290 was explored using RNA immunoprecipitation, qRT-PCR, and Western blot assays.Results The expression of circRNA_100290 was found significantly upregulated in GC cells and 102 GC tissues, high expression of circRNA_100290 in GC was closely related to Borrmann’s types, lymph node metastasis and tumor-node-metastasis staging. In vitro, knockdown of circRNA_100290 in AGS and HGC-27 cells significantly inhibited cell proliferation, migration, and invasion. Mechanistically, dual-luciferase reporter assay confirmed a direct binding between circRNA_100290 and miR-29b-3p, which targets ITGA11, an oncogene which is closely related to epithelial–mesenchymal transition (EMT). In addition, EIF4A3, one of RNA binding proteins (RBPs), could inhibit the formation of circRNA_100290 via enriching flanking sites of circRNA_100290. Low expression of EIF4A3 in GC was related to a worse prognosis.Conclusions Elevated circRNA_100290 in GC promotes cell proliferation, invasion and EMT via miR-29b-3p/ITGA11 axi and might be regulated by EIF4A3. CircRNA_100290 might be a promising biomarker and target for GC therapy.

scholarly journals Heat Shock Protein A12B Protects Vascular Endothelial Cells Against Sepsis-Induced Acute Lung Injury in Mice

2017 ◽  
Vol 42 (1) ◽  
pp. 156-168 ◽  
Author(s):  
Yi Chen ◽  
Lei Wang ◽  
Qiuxiang Kang ◽  
Xu Zhang ◽  
Guifang Yu ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Caspase 3 ◽  
Myeloperoxidase Activity ◽  
Expression Levels ◽  
Erk Phosphorylation ◽  
Tnf Α ◽  
Nasal Inhalation ◽  
Group A

Background: Pulmonary endothelial injury is a critical process in the pathogenesis of acute lung injury (ALI) during sepsis. Heat shock protein A12B (HSPA12B) is mainly expressed in endothelial cells and protects against several harmful factors. However, the effects of HSPA12B in sepsis-induced ALI and its potential mechanisms of action remain unclear. Methods: For in vivo experiments, C57BL/6 mice were randomly divided into four groups (n=15): a sham operation group, a cecal ligation and puncture (CLP) group, a HSPA12B siRNA-CLP group and a negative control (NC) siRNA-CLP group. The mice were treated by nasal inhalation of 2-OMe-modified HSPA12B siRNA or NC siRNA. Sepsis was induced by CLP. Samples were harvested 24 and 48 hours post-CLP surgery. Pathological changes and scoring of lung tissue samples were monitored using hematoxylin and eosin staining. Levels of pro-inflammatory cytokines (e.g., interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and IL-6) and myeloperoxidase activity in bronchoalveolar lavage fluid were analyzed by ELISA. Pulmonary edema was assessed using a wet-to-dry weight ratio. Neutrophils and alveolar macrophages were counted using flow cytometry. Pulmonary endothelial cell apoptosis was detected by TUNEL staining. Expression levels of MAPK family signaling molecules and caspase-3 were measured by Western blot analysis. In addition, 7-day survival was recorded. For in vitro experiments, human umbilical vein endothelial cells were pre-transfected with HSPA12B siRNA or pIRES2-EGFP-HSPA12B-Flag plasmid and treated with lipopolysaccharide; subsequently, the expression levels of MAPK family signaling molecules and caspase-3 were measured by Western blotting. Results: Nasal inhalation of nano-polymer-encapsulated HSPA12B siRNA specifically downregulated mRNA and protein expression levels of HSPA12B in lung tissues. The administration of HSPA12B siRNA aggravated lung pathological injury, upregulated pro-inflammatory cytokine (e.g., IL-1β, TNF-α, and IL-6) expression, and increased myeloperoxidase activity, neutrophil infiltration, pulmonary edema, and pulmonary endothelial cell apoptosis. Additionally, HSPA12B knockdown worsened survival after CLP surgery. The potential protective mechanisms of HSPA12B may involve the inhibition of ERK phosphorylation and caspase-3 activation in vivo and in vitro. Conclusion: HSPA12B protected against sepsis-induced ALI. The potential mechanism may be partly due to the inhibition of ERK phosphorylation and caspase-3 activation. These findings provide a potential therapeutic target for treating sepsis.

scholarly journals Pearl extract protects HaCaT cells from UV radiation-induced apoptosis through mitochondrial pathway regulation

2020 ◽  
Author(s):  
Zhixiong Chen ◽  
jing wang ◽  
Anquan Yang ◽  
Lihua Zhang ◽  
Yaojia Lu ◽  
...  
Keyword(s):  
Protein Expression ◽  
Caspase 3 ◽  
Hacat Cells ◽  
Caspase 9 ◽  
Expression Levels ◽  
Protein Levels ◽  
Bax Protein ◽  
Reactive Oxygen Species Content ◽  
Tnf Α ◽  
Induced Apoptosis

Abstract Background: Previous studies have demonstrated that pearl extract (PE) promotes wound healing and skin whitening. However, it remains unclear whether PE can inhibit ultraviolet (UV)-photodamage in HaCaT cells. In this study, an in vitro photoaging cell model was established to observe the effect of PE on UV-induced damage and the apoptosis of HaCaT cells. The aim of this study was to provide a reference for the future development of natural sunscreens.Results: PE concentrations of 0.1 and 1 μg/mL were considered the most effective and safe concentrations. Compared to that in the control group, superoxide dismutase and glutathione peroxidase activity in the photoaging group was significantly reduced, whereas malondialdehyde and reactive oxygen species content, along with tumour necrosis factor-alpha (TNF-α) and interleukin (IL)-10 mRNA and protein levels, were markedly increased. In contrast, Bcl-2 protein expression was significantly decreased, whereas caspase-3, caspase-9, and Bax protein expression levels were significantly increased. Compared to that in the photoaging group, HaCaT cell proliferation was significantly increased in the PE group. Both PE concentrations significantly increased superoxide dismutase and glutathione peroxidase activity in cells, reduced malondialdehyde and reactive oxygen species content, decreased TNF-α and IL-10 mRNA expression in cells, and reduced TNF-α and IL-10 protein levels in the supernatant. Additionally, Bcl-2 protein expression levels were significantly increased, whereas caspase-3, caspase-9, and Bax protein expression levels were significantly reduced by PE treatment.Conclusions: PE can inhibit UV-induced apoptosis by inhibiting mitochondria-mediated apoptosis and regulating TNF-α and IL-10 expression.

scholarly journals Glycolytic Reprogramming in Silica-Induced Lung Macrophages and Silicosis Reversed by Ac-SDKP Treatment

2021 ◽  
Vol 22 (18) ◽  
pp. 10063
Author(s):  
Na Mao ◽  
Honghao Yang ◽  
Jie Yin ◽  
Yaqian Li ◽  
Fuyu Jin ◽  
...  
Keyword(s):  
Pulmonary Fibrosis ◽  
Western Blot ◽  
Macrophage Activation ◽  
Specific Mechanism ◽  
Key Enzymes ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Lung Macrophages ◽  
Tnf Α ◽  
Metabolic Feature

Glycolytic reprogramming is an important metabolic feature in the development of pulmonary fibrosis. However, the specific mechanism of glycolysis in silicosis is still not clear. In this study, silicotic models and silica-induced macrophage were used to elucidate the mechanism of glycolysis induced by silica. Expression levels of the key enzymes in glycolysis and macrophage activation indicators were analyzed by Western blot, qRT-PCR, IHC, and IF analyses, and by using a lactate assay kit. We found that silica promotes the expression of the key glycolysis enzymes HK2, PKM2, LDHA, and macrophage activation factors iNOS, TNF-α, Arg-1, IL-10, and MCP1 in silicotic rats and silica-induced NR8383 macrophages. The enhancement of glycolysis and macrophage activation induced by silica was reduced by Ac-SDKP or siRNA-Ldha treatment. This study suggests that Ac-SDKP treatment can inhibit glycolytic reprogramming in silica-induced lung macrophages and silicosis.

The P-MAPA immunomodulator partially prevents apoptosis induced by Zika virus infection in THP-1 cells

2020 ◽  
Vol 21 ◽  
Author(s):  
Morganna C. Lima ◽  
Elisa A. N. Azevedo ◽  
Clarice N. L. de Morais ◽  
Larissa I. O. de Sousa ◽  
Bruno M. Carvalho ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Death ◽  
Virus Infection ◽  
Virus Replication ◽  
Zika Virus ◽  
Mammalian Cells ◽  
Caspase 3 ◽  
Neurological Complications ◽  
Zika Virus Infection

Background: Zika virus is an emerging arbovirus of global importance. ZIKV infection is associated with a range of neurological complications such as the Congenital Zika Syndrome and Guillain Barré Syndrome. Despite the magnitude of recent outbreaks, there is no specific therapy to prevent or to alleviate disease pathology. Objective: To investigate the role of P-MAPA immunomodulator in Zika-infected THP-1 cells. Methods: THP-1 cells were subjected at Zika virus infection (Multiplicity of Infection = 0.5) followed by treatment with P-MAPA for until 96 hours post-infection. After that, the cell death was analyzed by annexin+/ PI+ and caspase 3/ 7+ staining by flow cytometry. In addition, the virus replication and cell proliferation were accessed by RT-qPCR and Ki67 staining, respectively. Results: We demonstrate that P-MAPA in vitro treatment significantly reduces Zika virus-induced cell death and caspase-3/7 activation on THP-1 infected cells, albeit it has no role in virus replication and cell proliferation. Conclusions: Our study reveals that P-MAPA seems to be a satisfactory alternative to inhibits the effects of Zika virus infection in mammalian cells.

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