MicroRNA-298 determines the radio-resistance of colorectal cancer cells by directly targeting human dual-specificity tyrosine(Y)-regulated kinase 1A

BACKGROUND Radiotherapy stands as a promising therapeutic modality for colorectal cancer(CRC);yet,the formidable challenge posed by radio-resistance significantly undermines its efficacy in achieving CRC remission.AIM To elucidate the role played by microRNA-298(miR-298)in CRC radio-resistance.METHODS To establish a radio-resistant CRC cell line,HT-29 cells underwent exposure to 5 gray ionizing radiation that was followed by a 7-d recovery period.The quantification of miR-298 levels within CRC cells was conducted through quantitative RT-PCR,and protein expression determination was realized through Western blotting.Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and proliferation by clonogenic assay.Radio-induced apoptosis was discerned through flow cytometry analysis.RESULTS We observed a marked upregulation of miR-298 in radio-resistant CRC cells.MiR-298 emerged as a key determinant of cell survival following radiation exposure,as its overexpression led to a notable reduction in radiation-induced apoptosis.Intriguingly,miR-298 expression exhibited a strong correlation with CRC cell viability.Further investigation unveiled human dual-specificity tyrosine(Y)-regulated kinase 1A(DYRK1A)as miR-298’s direct target.CONCLUSION Taken together,our findings underline the role played by miR-298 in bolstering radio-resistance in CRC cells by means of DYRK1A downregulation,thereby positioning miR-298 as a promising candidate for mitigating radioresistance in CRC.

KAT7/HMGN1 signaling epigenetically induces tyrosine phosphorylation-regulated kinase 1A expression to ameliorate insulin resistance in Alzheimer’s disease

BACKGROUND Epidemiological studies have revealed a correlation between Alzheimer’s disease(AD)and type 2 diabetes mellitus(T2D).Insulin resistance in the brain is a common feature in patients with T2D and AD.KAT7 is a histone acetyltransferase that participates in the modulation of various genes.AIM To determine the effects of KAT7 on insulin patients with AD.METHODS APPswe/PS1-dE9 double-transgenic and db/db mice were used to mimic AD and diabetes,respectively.An in vitro model of AD was established by Aβstimulation.Insulin resistance was induced by chronic stimulation with high insulin levels.The expression of microtubule-associated protein 2(MAP2)was assessed using immunofluorescence.The protein levels of MAP2,Aβ,dual-specificity tyrosine phosphorylation-regulated kinase-1A(DYRK1A),IRS-1,p-AKT,total AKT,p-GSK3β,total GSK3β,DYRK1A,and KAT7 were measured via western blotting.Accumulation of reactive oxygen species(ROS),malondialdehyde(MDA),and SOD activity was measured to determine cellular oxidative stress.Flow cytometry and CCK-8 assay were performed to evaluate neuronal cell death and proliferation,respectively.Relative RNA levels of KAT7 and DYRK1A were examined using quantitative PCR.A chromatin immunoprecipitation assay was conducted to detect H3K14ac in DYRK1A.RESULTS KAT7 expression was suppressed in the AD mice.Overexpression of KAT7 decreased Aβaccumulation and MAP2 expression in AD brains.KAT7 overexpression decreased ROS and MDA levels,elevated SOD activity in brain tissues and neurons,and simultaneously suppressed neuronal apoptosis.KAT7 upregulated levels of p-AKT and p-GSK3βto alleviate insulin resistance,along with elevated expression of DYRK1A.KAT7 depletion suppressed DYRK1A expression and impaired H3K14ac of DYRK1A.HMGN1 overexpression recovered DYRK1A levels and reversed insulin resistance caused by KAT7 depletion.CONCLUSION We determined that KAT7 overexpression recovered insulin sensitivity in AD by recruiting HMGN1 to enhance DYRK1A acetylation.Our findings suggest that KAT7 is a novel and promising therapeutic target for the resistance in AD.

Minibrain-related kinase/dual-specificity tyrosine-regulated kinase 1B implication in stem/cancer stem cells biology

Dual-specificity tyrosine phosphorylation-regulated kinase 1B(DYRK1B),also known as minibrain-related kinase(MIRK)is one of the best functionally studied members of the DYRK kinase family.DYRKs comprise a family of protein kinases that are emerging modulators of signal transduction pathways,cell proliferation and differentiation,survival,and cell motility.DYRKs were found to participate in several signaling pathways critical for development and cell homeostasis.In this review,we focus on the DYRK1B protein kinase from a functional point of view concerning the signaling pathways through which DYRK1B exerts its cell type-dependent function in a positive or negative manner,in development and human diseases.In particular,we focus on the physiological role of DYRK1B in behavior of stem cells in myogenesis,adipogenesis,spermatogenesis and neurogenesis,as well as in its pathological implication in cancer and metabolic syndrome.Thus,understanding of the molecular mechanisms that regulate signaling pathways is of high importance.Recent studies have identified a close regulatory connection between DYRK1B and the hedgehog(HH)signaling pathway.Here,we aim to bring together what is known about the functional integration and cross-talk between DYRK1B and several signaling pathways,such as HH,RAS and PI3K/mTOR/AKT,as well as how this might affect cellular and molecular processes in development,physiology,and pathology.Thus,this review summarizes the major known functions of DYRK1B kinase,as well as the mechanisms by which DYRK1B exerts its functions in development and human diseases focusing on the homeostasis of stem and cancer stem cells.

Fermented Barley Extracts with Lactobacillus plantarum dy-1 Rich in Vanillic Acid Modulate Glucose Consumption in Human HepG2 Cells

Objective To investigate the effect of fermented barley extracts with Lactobacillus plantarum dy-1（LFBE） for modulating glucose consumption in HepG2 cells via miR-212 regulation. Methods Hepatocellular carcinoma（HepG2） cells were treated with palmitate. After 12 h, palmitate-induced HepG2 cells were treated with LFBE and its main components. Changes in glucose consumption, proinflammatory cytokine secretion, and miRNA-212 expression in HepG2 cells was observed. Results Treatment with LFBE rich in vanillic acid（VA） increased glucose consumption and reduced proinflammatory cytokine secretion in HepG2 cells. LFBE and VA normalized the upregulation of miR-212, which led to the upregulation of dual-specificity phosphatase-9（DUSP9）, a direct target of miR-212, at both protein and mR NA levels. Downregulation of miR-212 markedly increased glucose consumption and reduced proinflammatory cytokine secretion by enhancing DUSP9 expression. Conclusion The results showed the benefit of LFBE and miR-212 downregulation in modulating glucose consumption and reducing proinflammatory cytokine secretion by targeting DUSP9. VA in LFBE was a strong regulator of palmitate-induced abnormal glucose consumption in HepG2 cells and can be a primary mediator.

Study on effect of imbalance of TRAF-6, IRAK-1 and NALP3 inflammatory factors in patients with gouty arthritis

Objective:To explore the effect of imbalance of tumor necrosis factor receptor related factor-6(TRAF-6),interleukin 1 receptor associated kinase-1(IRAK-1)and neutrophil alkaline phosphatase-3(NALP3)in patients with gouty arthritis.Methods:The retrospective experiment was conducted on 105 patients with gouty arthritis admitted to our hospital(47 patients with acute onset and 58 patients with remission,namely group A and group B);meanwhile,another 61 healthy volunteers were selected for control,namely group C.The enrolling of the three groups was dated from May 2017 to May 2018,and TRAF-6,IRAK-1 and NALP3 of all subjects were tested through real-time fluorescence quantification(RT-PCR),and the correlation between the three inflammatory factors and gouty arthritis was compared.Results:1)Through treatment,ESR,BUA and total addiment in group A and B were higher than those in group C,among which the three indicators in group A were higher than those in group B(P<0.05),while CRP was lower than that of group C,and the two indicators in group A were lower than those in group B(P<0.05).2)There was no significant difference in the relative expression of TRAF-6 mRNA between group A and group B before treatment(P>0.05),significantly lower than group C(P<0.05);the above indicators of group A and group B were improved to some extent after treatment,but group A was still lower than group B(P<0.05),and the degree of improvement of group A was also lower than that of group C(P<0.05),while the degree of improvement of group B was not significantly different from that of group C(P>0.05).3)The relative expression level of IRAK-1mRNA in group A and group B before treatment showed no significant difference(P>0.05),but was also lower than that in group C(P<0.05).The relative expression level of IRAK-1mRNA in group A and group B increased to some extent after treatment,with group A significantly lower than group C(P<0.05),and group B showed no significant difference compared with group C(P>0.05).4)The relative expression level of NALP-3 mRNA in group A and group B showed no significant difference(P>0.05)before treatment,significantly higher than that in group C(P<0.05);the relative expression of NALP-3 mRNA in group A was not significantly decreased(P>0.05)after treatment,while that in group B was significantly decreased after treatment(P<0.05),indicating significant different compared with group A and group C.5)There was no correlation between)TRAF-6,ESR,CRP and total addiment(P>0.05);IRAK-1 was negatively correlated with CRP,BUA and total addiment(P<0.05);NALP-3 was negatively correlated with ESR and CRP(P<0.05).Conclusion:TRAF-6,IRAK-1 and NALP-3 are all under abnormal expression in the developing of new gouty arthritis,acting as important participants in promoting the occurrence,development and outcome of illness states,so the intervening measures should be taken.

Effects of tetrandrine on phenotypic modulation of vascular smooth muscle cells and expression of p38 MAPK as well as MKP-1 after intimal injury of rabbit carotid arteries

Objective： To study the effects of tetrandrine （Tet） on phenotypic modulation of vascular smooth muscle cells （VSMCs） and expression of p38 mitogen-activated protein kinase （p38MAPK） as well as mitogen-activated protein kinase phosphatase-1 （MKP-1） after vascular intimal injury. Methods： HE staining was used to analyze vascular morphology of sham-injured group, injured group and Tet-treated group at day 28. lmmunohistochemistry, Western blot and RT-PCR were respectively used to detect the expression change of smooth muscle a-actin （SMa-actin）, proliferation cell nuclear antigen （PCNA）, p38MAPK and MKP-1 of injured group and Tet group at days 7, 14 and 28 after balloon injury. Results： ① All layers of vascular wall in sham-injured group were intact at day 28. The neointimal area was significantly increased and the lumen area notably decreased in injured group at day 28. The neointimal proliferation in Tet treated group was less than that in injured group, and the lumen area of Tet group was significantly increased than that of injured group at day 28. ②Compared with the injured group, the expression of SMa-actin, PCNA, p38MAPK and MKP-1 of vascular wall in Tet group was no difference, and the neointimal proliferation condition was also basically as same as injured group at day 7 after injury. The expression of PCNA and p38MAKP in Tet group was obviously lower than that in injured group, and the expression of MKP-1 in Tet group was obviously higher than that in injured group at days 14 and 28 after injury. The expression of SMa-actin in Tet group was slightly higher than that in injured group at days 14 and 28 after injury. Conclusions： Tet could reduce neointimal proliferation by inhibiting VSMCs phenotypic modulation and p38MAPK signaling transduction pathway as well as its down regulation.

Dual-specificity Phosphatase 1 Deficiency Induces Endometrioid Adenocarcinoma Progression via Activation of Mitogen-activated Protein Kinase/Extracellular Signal-regulated Kinase Pathway

Background： Previously, we reported that dual-specificity adenocarcinoma （EEA）. However, the role of DUSP1 medroxyprogesterone （MPA） are still unclear. phosphatase I （DUSPI） was differentially expressed in endometrioid in EEA progression and the relationship between DUSPI and Methods： The expression of DUSPI in EEA specimens was detected by immunohistochemical analysis. The effect of DUSPI on cell proliferation was analyzed by Cell Counting Kit 8 and colony formation assay, and cell migration was analyzed by transwell assay. MPA-induced DUSPI expression in EEA cells was measured by Western blot. Results： DUSPI expression was deficient in advanced International Federation of Gynecology and Obstetrics stage, high-grade and myometrial invasive EEA. In EEA cell lines （HeclA, Hecl B, RL952, and Ishikawa）, the DUSP1 expression was substantially higher in lshikawa cells than in other cell lines （P 〈 0.05）. Knockdown ofDUSP I promoted lshikawa cells proliferation, migration, and activation of mitogen-activated protein kinases/extracellular signal-regulated kinase （MAPK/Erk） pathway. MPA-induced DUSP1 expression and inhibited MAPK/Erk pathway in Ishikawa cells. Conclusions： Our data suggest that DUSP1 deficiency promotes EEA progression via MAPK/Erk pathway, which may be reversed by MPA, suggesting that DUSP I may serve as a potential therapeutic target for the treatment of EEA.

Let-7a gene knockdown protects against cerebral ischemia/reperfusion injury

The micro RNA（mi RNA） let-7 was one of the first mi RNAs to be discovered, and is highly conserved and widely expressed among species. let-7 expression increases in brain tissue after cerebral ischemia/reperfusion injury; however, no studies have reported let-7 effects on nerve injury after cerebral ischemia/reperfusion injury. To investigate the effects of let-7 gene knockdown on cerebral ischemia/reperfusion injury, we established a rat model of cerebral ischemia/reperfusion injury. Quantitative reverse transcription-polymerase chain reaction demonstrated that 12 hours after cerebral ischemia/reperfusion injury, let-7 expression was up-regulated, peaked at 24 hours, and was still higher than that in control rats after 72 hours. Let-7 gene knockdown in rats suppressed microglial activation and inflammatory factor release, reduced neuronal apoptosis and infarct volume in brain tissue after cerebral ischemia/reperfusion injury. Western blot assays and luciferase assays revealed that mitogen-activated protein kinase phosphatase-1（MKP1） is a direct target of let-7. Let-7 enhanced phosphorylated p38 mitogen-activated protein kinase（MAPK） and c-Jun N-terminal kinase（JNK） expression by down-regulating MKP1. These findings suggest that knockdown of let-7 inhibited the activation of p38 MAPK and JNK signaling pathways by up-regulating MKP1 expression, reduced apoptosis and the inflammatory reaction, and exerted a neuroprotective effect following cerebral ischemia/reperfusion injury.

Imbalanced expression of mitogen-activated protein kinase phosphatase-1 and phosphorylated extracellular signal-regulated kinases in lung squamous cell carcinoma

Objective： Mitogen-activated protein kinases （MAPKs） are correlated with a more malignant phenotype in many cancers. This study was designed to evaluate the predictive value of the expression of MAPK phosphatase-1 （MKP-1） and phosphorylated extracellular signal-regulated kinase 1/2 （p-ERKl/2）, as the key regulatory mechanism of the MAPKs, in lung squamous cell carcinoma （SCC）. Methods： We assessed the expressions of MKP-1 and p-ERK1/2 in twenty subjects at different differentiation degree of SCC and five normal lungs by immunohistochemistry and real-time reverse transcriptase polymerase chain reaction （RT-PCR） analysis. Results： Immunohistochemistry and real-time RT-PCR assay showed that the expression of MKP-1 was gradually decreased as tissue type went from normal lung tissues to increasingly undifferentiated carcinoma, and it was negatively correlated with tumor differentiation （P〈0.01）. However, the expression of p-ERK1/2 or ERKl/2 was gradually increased as tissue type went from normal lung tissues to increasingly undifferentiated carcinoma, and it was positively correlated with tumor differentiation （P〈0.01）. Conclusions： Our data indicates the relevance of MKP-1 and p-ERK1/2 in SCC as a potential positive and negative prognostic factor. The imbalanced expression of MKP-1 and p-ERKl/2 may play a role in the development of SCC and these two molecules may be new targets for the therapy and prognosis of SCC.

Effects of Glucocorticoid?Induced Transcript 1 Gene Deficiency on Glucocorticoid Activation in Asthmatic Mice

Background: Glucocorticoid (GC) is the first?line therapy for asthma, but some asthmatics are insensitive to it. Glucocorticoid?induced transcript 1 gene (GLCCI1) is reported to be associated with GCs efficiency in asthmatics, while its exact mechanism remains unknown. Methods: A total of 30 asthmatic patients received fluticasone propionate for 12 weeks. Forced expiratory volume in 1 s (FEV1) and GLCCI1 expression were detected. Asthma model was constructed in wild?type and GLCCI1 knockout (GLCCI1?/?) mice. Glucocorticoid receptor (GR) and mitogen?activated protein kinase phosphatase 1 (MKP?1) expression were detected by polymerase chain reaction and Western blotting (WB). The phosphorylation of p38 mitogen?activated protein kinase (MAPK) was also detected by WB. Results: In asthmatic patients, the change of FEV1 was well positively correlated with change of GLCCI1 expression (r = 0.430, P = 0.022). In animal experiment, GR and MKP?1 mRNA levels were significantly decreased in asthmatic mice than in control mice (wild?type: GR: 0.769 vs. 1.000, P = 0.022; MKP?1: 0.493 vs. 1.000, P < 0.001. GLCCI1?/?: GR: 0.629 vs. 1.645, P < 0.001; MKP?1: 0.377 vs. 2.146, P < 0.001). Hydroprednisone treatment significantly increased GR and MKP?1 mRNA expression levels than in asthmatic groups; however, GLCCI1?/?.asthmatic mice had less improvement (wild?type: GR: 1.517 vs. 0.769, P = 0.023; MKP?1: 1.036 vs. 0.493, P = 0.003. GLCCI1?/?: GR: 0.846 vs. 0.629, P = 0.116; MKP?1: 0.475 vs. 0.377, P = 0.388). GLCCI1?/? asthmatic mice had more obvious phosphorylation of p38 MAPK than wild?type asthmatic mice (9.060 vs. 3.484, P < 0.001). It was still higher even though after hydroprednisone treatment (6.440 vs. 2.630, P < 0.001). Conclusions: GLCCI1 deficiency in asthmatic mice inhibits the activation of GR and MKP?1 and leads to more obvious phosphorylation of p38 MAPK, leading to a decremental sensitivity to GCs.

DUSP1 Blocks autophagy-dependent ferroptosis in pancreatic cancer

Ferroptosis is a oxidative damage-dependent form of regulated cell death that has become an emerging target for disease prevention and treatment.Here,we show that dual-specificity phosphatase 1(DUSP1),a phosphatase playing multiple roles in stress-signaling pathways,is a new repressor of ferroptosis in human pancreatic cancer cells.Several classical ferroptosis activators(eg,erastin and RSL3)induce the expression of DUSP1,but not other members of DUSP,which depends on extracellular signal-regulated protein kinases 1 and 2(ERK1/2).Moreover,shRNA-mediated DUSP1 knockdown increases the anticancer activity of ferroptosis activators in pancreatic cancer cells through activating lipid peroxidation in vitro and in vivo.Importantly,DUSP1-mediated autophagy is responsible for lipid peroxidation-mediated ferroptotic cell death.Thus,the DUSP1-related ferroptotic pathway may represent a potential target for therapeutic intervention in pancreatic cancer.

Electrochemical inactivation of cyanobacteria and microcystin degradation using a boron-doped diamond anode-A potential tool for cyanobacterial bloom control

Cyanobacterial blooms are global phenomena that can occur in calm and nutrient-rich （eutrophic） fresh and marine waters. Human exposure to cyanobacteria and their biologically active products is possible during water sports and various water activities, or by ingestion of contaminated water. Although the vast majority of harmful cyanobacterial products are confined to the interior of the cells, these are eventually released into the surrounding water following natural or artificially induced cell death. Electrochemical oxidation has been used here to damage cyanobacteria to halt their proliferation, and for microcystin degradation under in-vitro conditions. Partially spent Jaworski growth medium with no addition of supporting electrolytes was used. Electrochemical treatment resulted in the cyanobacterial loss of cell-buoyancy regulation, cell proliferation arrest, and eventual cell death. Microcystin degradation was studied separately in two basic modes of treatment： batch-wise flow, and constant flow, for electrolytic-cell exposure. Batch-wise exposure simulates treatment under environmental conditions, while constant flow is more appropriate for the study of boron-doped diamond electrode efficacy under laboratory conditions. The effectiveness of microcystin degradation was established using high-performance liquid chromatography-photodiode array detector analysis, while the biological activities of the products were estimated using a colorimetric protein phosphatase-1 inhibition assay. The results indicate potential for the application of electro-oxidation methods for the control of bloom events by taking advantage of specific intrinsic ecological characteristics of bloom-forming cyanobacteria. The applicability of the use of boron-doped diamond electrodes in remediation of water exposed to cyanobacteria bloom events is discussed.