Names Targeting p65 to Inhibit Cas3 Transcription by Onjisaponin B for Radiation Damage Therapy in p65+/- Mice

Abstract Background: p65 is activated following radiation injury. The formation of p65 is regulated by Onjisaponin B (OB) in Alzheimer's disease models. In addition, there is a binding site for p65 in the promoter region of CAS3. In the present study, the use of OB as an intervention to modulate p65/Cas3 following radiation injury was studied.Methods: Cellular and animal experiments, immunofluorescence, HE staining, Western blotting, qRT-PCR, comet and DNA ladder assays, and flow cytometry were used to confirm the expression of p65 and Cas3.Results: The results demonstrated that if the expression of p65 was silenced in V79 and TC cells, OB did not significantly inhibit the activation of p65 or Cas3 following irradiation, or significantly inhibit the phosphorylation of p65 and its transfer into the nucleus. Overexpression of p65 in V79 and MTEC-1 cells resulted in OB significantly inhibiting the activation of p65 and Cas3, and the phosphorylation and translocation of p65 into the nucleus. In p65+/- mice, expression of the p65 gene was knocked down, leading to increased tissue apoptosis and inflammation, and serious tissue pathological changes. The inhibition of p65 activation by OB after exposure to radiation was not apparent in the thymus, but it was in the lung, indicating that OB has a regulatory effect on endogenous p65.Conclusions:In summary, OB interfered with radiation injury by targeting and regulating p65/Cas3. Therefore, it was confirmed that p65 is an important target molecule for the treatment of radiation injury.

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Cytotoxic Effect of RecombinantMycobacterium tuberculosisCFP-10/ESAT-6 Protein on the Crucial Pathways of WI-38 Cells

Journal of Biomedicine and Biotechnology ◽

10.1155/2009/917084 ◽

2009 ◽

Vol 2009 ◽

pp. 1-10 ◽

Cited By ~ 7

Author(s):

Kun-Nan Tsai ◽

Err-Cheng Chan ◽

Tsung-Yeh Tsai ◽

Kuei-Tien Chen ◽

Chun-Yu Chen ◽

...

Keyword(s):

Focal Adhesion ◽

Microarray Data ◽

Cytotoxic Effect ◽

Time Course ◽

Essential Role ◽

Topological Properties ◽

Regulatory Effect ◽

Qrt Pcr ◽

Focal Adhesion Pathway ◽

Pathway Databases

To unravel the cytotoxic effect of the recombinant CFP-10/ESAT-6 protein (rCFES) on WI-38 cells, an integrative analysis approach, combining time-course microarray data and annotated pathway databases, was proposed with the emphasis on identifying the potentially crucial pathways. The potentially crucial pathways were selected based on a composite criterion characterizing the average significance and topological properties of important genes. The analysis results suggested that the regulatory effect of rCFES was at least involved in cell proliferation, cell motility, cell survival, and metabolisms of WI-38 cells. The survivability of WI-38 cells, in particular, was significantly decreased to 62% with 12.5 μMrCFES. Furthermore, the focal adhesion pathway was identified as the potentially most-crucial pathway and 58 of 65 important genes in this pathway were downregulated by rCFES treatment. Using qRT-PCR, we have confirmed the changes in the expression levels of LAMA4, PIK3R3, BIRC3, and NFKBIA, suggesting that these proteins may play an essential role in the cytotoxic process in the rCFES-treated WI-38 cells.

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lncRNA MIR22HG-Derived miR-22-5p Enhances the Radiosensitivity of Hepatocellular Carcinoma by Increasing Histone Acetylation Through the Inhibition of HDAC2 Activity

Frontiers in Oncology ◽

10.3389/fonc.2021.572585 ◽

2021 ◽

Vol 11 ◽

Author(s):

Qiao Jin ◽

Hao Hu ◽

Siqi Yan ◽

Long Jin ◽

Yuliang Pan ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Histone Acetylation ◽

Chromatin Immunoprecipitation ◽

Promoter Region ◽

Animal Experiments ◽

Primary Hepatocellular Carcinoma ◽

Acetylation Level ◽

Expression Levels

BackgroundWith the development of radiotherapy technology, radiotherapy has been increasingly used to treat primary hepatocellular carcinoma (HCC). However, due to radioresistance and the intolerance of the adjacent organs to radiation, the effects of radiotherapy are often unsatisfactory. Therefore, it is necessary to study radiosensitization in HCC.MethodA microarray was used to analyze the genes that were significantly associated with radiosensitivity. HCC cells, HepG2 and MHCC97H, were subjected to radiation in vitro. Real-time PCR was performed to determine MIR22HG (microRNA22 host gene) and miR-22-5p expression levels. Western blotting was performed to determine histone expression levels. A histone deacetylase (HDAC) whole cell assay was used to determine the activity of HDAC2. MTT, colony formation, 5-ethynyl-2′-deoxyuridine, and wound healing assays were performed to examine the function of MIR22HG and miR-22-5p in cellular radiosensitivity. Chromatin immunoprecipitation-PCR was used to confirm that HDAC2 affects the acetylation level of the MIR22HG promoter region. Finally, animal experiments were performed to demonstrate the in vivo effect of MIR22HG on the radiosensitivity of hepatoma.ResultsIrradiation can up-regulate MIR22HG expression and down-regulate HDAC2 expression. Inhibition of HDAC2 expression promotes histone acetylation in the MIR22HG promoter region and up-regulates MIR22HG expression. MIR22HG can increase radiosensitivity via miR-22-5p in HCC.ConclusionInhibition of HDAC2 expression promotes histone acetylation in the MIR22HG promoter region, thereby up-regulating the expression of MIR22HG and promoting the production of miR-22-5p, and ultimately increasing the sensitivity of liver cancer radiotherapy.

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KCNQ1OT1 Knock Down Alleviates Hyperosmlity Induced Dry Eye Disease via Inhibiting NLRP3 Inflammsome Activation and IL-1β Release

10.21203/rs.3.rs-721954/v1 ◽

2021 ◽

Author(s):

Xuran Li ◽

Yanyan Zhang ◽

Xinyue Li ◽

Yu Yu ◽

Xin Jin ◽

...

Keyword(s):

Cell Migration ◽

Dry Eye ◽

Noncoding Rnas ◽

Dry Eye Disease ◽

Eye Disease ◽

Pathological Changes ◽

Scopolamine Hydrobromide ◽

Qrt Pcr ◽

Age Related ◽

Set Up

Abstract Background. Long noncoding RNAs (lncRNAs) have been indicated that participate in inflammatory diaeases and age related diseases by regulating physiological and pathological processes directly or indirectly. Evidences have indicated lncRNAs might be involving in dry eye disease which can cause pathological changes of cornea and conjunctiva in ocular surface though inflammatory response. KCNQ1OT1 is a novel lncRNA, whose function remains totally unclear.Methods. qRT-PCR was performed to detect the expression of KCNQ1OT1, miR-214 and inflammasome-related genes NLRP3, caspase1 and IL-1β. Western blotting was carried out to detect inflammasome-related markers. The dry eye disease (DED) model was set up by scopolamine hydrobromide though subcutaneous injection. Bioinformatics was used to predict and validate the interaction between KCNQ1OT1 and miR-214 as well caspase1 and miR-214.Results. KCNQ1OT1 was significantly up-regulated during the process of DED while miR-214 was contrarily down-regulated. Knockdown of KCNQ1OT1 promoted cornea epithelial cells migration and down-regulated inflammasome-related genes. It was also confirmed that KCNQ1OT1 directly interacted with miR-214. Meanwhile, miR-214 could bind to 3'UTR of caspase1 and therefore inhibited its expression. Furthermore, co-transfection of miR-214 inhibitor could rescue the down-regulation of cell migration induced by KCNQ1OT1 knockdown.

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Notoginsenoside R2 reduces Aβ25-35-induced neuronal apoptosis and inflammation via miR-27a/SOX8/β-catenin axis

Human & Experimental Toxicology ◽

10.1177/09603271211041996 ◽

2021 ◽

pp. 096032712110419

Author(s):

Yueqiang Hu ◽

Lin Wu ◽

Lingfei Jiang ◽

Ni Liang ◽

Xiaomin Zhu ◽

...

Keyword(s):

Cortical Neurons ◽

Neuronal Apoptosis ◽

Animal Experiments ◽

Panax Notoginseng ◽

Luciferase Reporter ◽

Beta Peptide ◽

Rat Cortical Neurons ◽

Cellular Apoptosis ◽

Apoptosis And Inflammation

Background: Alzheimer’s disease (AD) has affected numerous elderly individuals worldwide. Panax notoginseng has been shown to ameliorate AD symptoms, and notoginsenoside R2 is a key saponin identified in this plant. Purpose: In the current study, we aimed to explore whether notoginsenoside R2 could improve the prognosis of AD. Methods: Herein, primary rat cortical neurons were isolated and they were treated with amyloid beta-peptide (A β) 25–35 oligomers. Cellular apoptosis was examined via flow cytometry and Western blotting. miR-27a and SOX8 mRNA expression levels were quantified by quantitative reverse transcription-polymerase chain reaction. Furthermore, the interaction between miR-27a and SOX8 was investigated by utilizing a dual-luciferase reporter assay. Finally, an AD mouse model was established to validate the in vitro findings. Results: Notoginsenoside R2 alleviated A β25-35-triggered neuronal apoptosis and inflammation. During this process, miR-27a expression was decreased by notoginsenoside R2, and miR-27a negatively modulated SOX8 expression. Furthermore, activation of SOX8 upregulated β-catenin expression, thus suppressing apoptosis and neuroinflammation. Conclusions: Our animal experiments revealed that notoginsenoside R2 enhanced the cognitive function of AD mice and inhibited neuronal apoptosis. Notoginsenoside R2 ameliorated AD symptoms by reducing neuronal apoptosis and inflammation, thus suggesting a novel direction for AD pharmacotherapy.

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Overexpression of angiotensin-converting enzyme 2 by renin-angiotensin system inhibitors. Truth or myth? A systematic review of animal studies

Hypertension Research ◽

10.1038/s41440-021-00641-1 ◽

2021 ◽

Author(s):

Hisashi Kai ◽

Mamiko Kai ◽

Hiroshi Niiyama ◽

Norihito Okina ◽

Motoki Sasaki ◽

...

Keyword(s):

Angiotensin Converting Enzyme ◽

Control Level ◽

Animal Experiments ◽

Renin Angiotensin System ◽

Organ Damage ◽

Disease Models ◽

Converting Enzyme ◽

Angiotensin System ◽

Angiotensin Converting Enzyme 2 ◽

Renin Angiotensin

AbstractAngiotensin-converting enzyme 2 (ACE2) protects against organ damage in hypertension and cardiovascular diseases by counter regulating the renin-angiotensin system (RAS). ACE2 is also the receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Based on the claim that RAS inhibitors (RASIs) cause ACE2 overexpression in some animal experiments, concerns have arisen that RASIs may aggravate SARS-CoV-2 infection and coronavirus disease-2019 severity in RASI-treated patients. To achieve a comprehensive review, a systematic search of MEDLINE/PubMed was conducted regarding the effects of RASIs on tissue ACE2 mRNA/protein expression in healthy animals and animal models of human diseases. We identified 88 eligible articles involving 168 experiments in the heart, kidneys, lungs, and other organs. Three of 38 experiments involving healthy animals showed ACE2 expression greater than twice that of the control (overexpression). Among 102 disease models (130 experiments), baseline ACE2 was overexpressed in 16 models (18 experiments) and less than half the control level (repression) in 28 models (40 experiments). In 72 experiments, RASIs did not change ACE2 levels from the baseline levels of disease models. RASIs caused ACE2 overexpression compared to control levels in seven experiments, some of which were unsupported by other experiments under similar conditions. In 36 experiments, RASIs reversed or prevented disease-induced ACE2 repression, yielding no or marginal changes. Therefore, ACE2 overexpression appears to be a rare rather than common consequence of RASI treatment in healthy animals and disease models. Future studies should clarify the pathophysiological significance of RASI-induced reversal or prevention of ACE2 repression in disease models.

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Quanzhenyiqitang Reverses LPS-Induced Inflammation via Inhibiting PYK2/p38MAPK/HDAC2/CK2 Signaling Pathway in Rat Alveolar Macrophage

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2022/7857022 ◽

2022 ◽

Vol 2022 ◽

pp. 1-11

Author(s):

Ke-Qiang Chen ◽

Da-zhi Li ◽

Zhi-bin Chen ◽

Chuan-lin Zhang ◽

Bin-can Wang ◽

...

Keyword(s):

Lung Injury ◽

Pulmonary Disease ◽

Lung Inflammation ◽

Therapeutic Drug ◽

Chronic Obstructive ◽

Chronic Pulmonary Disease ◽

Pathological Changes ◽

Obstructive Pulmonary Disease ◽

Qrt Pcr ◽

Underlying Mechanisms

Chronic obstructive pulmonary disease (COPD) is a common chronic pulmonary disease with multiple etiologies and pathological changes. PYK2 expression is significantly increased in lipopolysaccharide-induced lung injury, but it mediates chronic lung inflammation. The mechanism of its occurrence remains unclear. Quanzhenyiqitang is often used in clinical treatment of COPD, so this study explored the mechanism of its treatment of lipopolysaccharide-induced lung injury. In this study, transfection, flow cytometry, QRT-PCR, and Western blotting methods were used to study the mechanism of Quanzhenyiqitang lipopolysaccharide-induced lung injury. The results showed that the mechanism of occurrence remains unclear. Our novel observations imply that the PYK2/p38MAPK/HDAC2/CK2 pathway is one of the fundamental underlying mechanisms that mediate the pathogenic progression of COPD, and Quanzhenyiqitang may be the therapeutic drug to prevent chronic inflammation and delay the progression of COPD by inhibiting PYK2 signaling pathways.

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Attenuation of Apoptosis Stimulating Protein of p53-1 (ASPP1) Associates with Therapy Failure in Acute Myeloid Leukemia (AML)

Blood ◽

10.1182/blood.v128.22.1556.1556 ◽

2016 ◽

Vol 128 (22) ◽

pp. 1556-1556

Author(s):

Marcus M. Schittenhelm ◽

Max Kaiser ◽

Gunnar Blumenstock ◽

Kerstin Maria Kampa-Schittenhelm

Keyword(s):

Bone Marrow ◽

Protein Expression ◽

Cell Lines ◽

Western Blotting ◽

Promoter Region ◽

Leukemia Cell ◽

Mrna Levels ◽

Primary Therapy ◽

Qrt Pcr ◽

Leukemia Cell Lines

Abstract ASPP1 belongs to a family of p53-binding proteins and enhances apoptosis by stimulation of p53-transactivation of selected proapoptotic target genes. It is preferentially expressed in hematopoietic stem cells (HSC) and together with p53 preserves the genomic integrity of the HSC pool. Consequently, attenuated expression of ASPP1 which is linked to methylation of the promoter region has been associated with malignant transformation and development of acute lymphoblastic leukemia and lymphomas. We now provide evidence that ASPP1 is highly altered in AML suggesting a role in leukemogenesis as well as therapy response. ASPP1 mRNA and protein expression levels of freshly isolated native patient samples (68) and healthy bone marrow donors (29) were determined by qRT-PCR and western immunoblotting. Statistical analyseswere performed. To explore implications of attenuated ASPP1 levels with regard to apoptosis induction and proliferation, ASPP1-expressing leukemia cell lines (MOLM14, Jurkat), native patient blasts or native bone marrow donor samples were stably silenced using a retroviral shRNA approach. Vice versa, ASPP1 was stably overexpressed in AML cell lines expressing per se low ASPP1 levels. Expression was thereby confirmed by qRT-PCR and western blotting. XTT viability and annexin V-based apoptosis assays were performed using standard chemotherapeutics in comparison to empty vector controls. Decitabine was used as an epigenetic sensitizer via hypomethylation of the promoter region. ASPP1 mRNA expression was found to be frequently and highly statistically significantly (p=0.001) attenuated in AML. Low ASPP1 mRNA levels thereby translated into attenuated protein expression. Retroviral ASPP1-interference lead to perturbed proliferation capacities (up to 3-fold increase) and attenuated apoptosis upon standard chemotherapeutics in leukemia cell lines as well as native leukemia blasts. As expected, overexpression of ASPP1 resulted in significantly attenuated proliferation and higher induction of apoptosis in all tested cell lines and patient blasts. Intriguingly, epigenetic therapy using the hypomethylating agent decitabine resulted in upregulation of ASPP1 expression in leukemia cells with originally low basal ASPP1 levels as confirmed by qRT-PCR and western blotting. Consequently, decitabine pretreatment sensitized these patient samples towards chemotherapy with a favorable proapoptotic overall efficacy compared to chemotherapy alone. Our results demonstrate that dysfunctional regulation of ASPP1 expression likely contributes to the biology of leukemogenesis and to primary therapy resistance in a subgroup of patients with acute leukemia and seems to be linked to hypermethylation. Prospective clinical studies are warranted to evaluate the roleas a biomarker for risk stratification in leukemia patients and for monitoring therapy responses. Disclosures No relevant conflicts of interest to declare.

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The PsMYB12L/PsDFR Module is Involved in Double-Color Formation in Paeonia Suffruticosa ‘Shima Nishiki’

10.21203/rs.3.rs-778866/v1 ◽

2021 ◽

Author(s):

Xinpeng Zhang ◽

Xu Han ◽

Mingyuan Zhao ◽

Xiaoyan Yu ◽

Cheng Li ◽

...

Keyword(s):

Dna Sequence ◽

Methylation Level ◽

Promoter Region ◽

Genomic Dna ◽

Cdna Sequence ◽

Regulatory Effect ◽

Paeonia Suffruticosa ◽

Color Formation ◽

Region Sequence ◽

Genomic Dna Sequence

Abstract Background: Paeonia suffruticosa ‘Shima Nishiki’ is a extremely precious double-color cultivar in the world because of its unique and attractive flower color. However, the underlying molecular mechanisms of its double-color formation have not been completely unravelled until now. In the present study, firstly, the full-length cDNA sequence, genomic DNA sequence, promoter region sequence of the PsDFR gene in the red and pink petals of the ‘Shima Nishiki’ cultivar were cloned and analyzed, respectively. Meanwhile, the methylation level of CpG island and promoter region of this gene in the red and pink petals was also measured. Moreover, the identification of regulatory effect of PsMYB114L/PsMYB12L and PsDFR was performed. Results: Here, we found that the full-length cDNA sequence, genomic DNA sequence, promoter region sequence of PsDFR were identical in the red and pink petals, respectively. There were some differences for the methylation level of this gene in the red and pink petals, but these differences were little and didn’t show obvious regularity. In addition, the regulatory effect of PsMYB12L and PsDFR was successfully identified. Conclusions: Based on these above results, we concluded that PsMYB12L regulating the differential expression of PsDFR may be a key reason for the double-color formation. These results will advance our understanding of the molecular regulatory mechanisms of double-color formation in P. suffruticosa ‘Shima Nishiki’.

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Differences in brain pathological changes between rotenone and 6-hydroxydopamine Parkinson’s disease models

Neural Regeneration Research ◽

10.4103/1673-5374.235076 ◽

2018 ◽

Vol 13 (7) ◽

pp. 1276 ◽

Cited By ~ 3

Author(s):

Lan-Xiang Liu ◽

Dan Du ◽

Zhan-Qiu Wang ◽

Yuan Fang ◽

Tao Zheng ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Disease Models ◽

Pathological Changes ◽

6 Hydroxydopamine

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Aberrant DNA Methylation-Mediated FOXF2 Dysregulation Is a Prognostic Risk Factor for Gastric Cancer

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.645470 ◽

2021 ◽

Vol 8 ◽

Author(s):

Cheng Zhang ◽

Yong-Zhi Li ◽

Dong-Qiu Dai

Keyword(s):

Gastric Cancer ◽

Dna Methylation ◽

Western Blot ◽

Methylation Level ◽

Promoter Region ◽

Target Genes ◽

Risk Group ◽

Rna Seq ◽

Qrt Pcr ◽

Aberrant Dna Methylation

Background: The prognosis of gastric cancer (GC) patients is poor. The effect of aberrant DNA methylation on FOXF2 expression and the prognostic role of FOXF2 methylation in GC have not yet been identified.Methods: The RNA-Seq and gene methylation HM450 profile data were used for analyzing FOXF2 expression in GC and its association with methylation level. Bisulfite sequencing PCR (BSP) was performed to measure the methylation level of the FOXF2 promoter region in GC cell lines and normal GES-1 cells. The cells were treated with the demethylation reagent 5-Aza-dC, and the mRNA and protein expression levels of FOXF2 were then measured by qRT-PCR and western blot assays. The risk score system from SurvivalMeth was calculated by integrating the methylation level of the cg locus and the corresponding Cox regression coefficient.Results: FOXF2 was significantly downregulated in GC cells and tissues. On the basis of RNA-Seq and Illumina methylation 450 data, FOXF2 expression was significantly negatively correlated with the FOXF2 methylation level (Pearson’s R = −0.42, p < 2.2e−16). The FOXF2 methylation level in the high FOXF2 expression group was lower than that in the low FOXF2 expression group. The BSP assay indicated that the methylation level of the FOXF2 promoter region in GC cell lines was higher than that in GES-1 cells. The qRT-PCR and western blot assay showed that FOXF2 mRNA and protein levels were increased in GC cells following treatment with 5-Aza-Dc. The methylation risk score model indicated that patients in the high risk group had poorer survival probability than those in the low risk group (HR = 1.84 (1.11–3.07) and p = 0.0068). FOXF2 also had a close transcriptional regulation network with four miRNAs and their corresponding target genes. Functional enrichment analysis of the target genes revealed that these genes were significantly related to several important signaling pathways.Conclusion: FOXF2 was downregulated due to aberrant DNA methylation in GC, and the degree of methylation in the promoter region of FOXF2 was related to the prognosis of patients. The FOXF2/miRNAs/target genes axis may play a vital biological regulation role in GC.

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