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At least 559 records · Page 31

MiR-615 inhibits cell proliferation, migration and invasion by targeting EGFR in human glioblastoma

Highlights: • MiR-615 is decreased in GBM. • MiR-615 is a tumor suppressor. • MiR-615 targets EGFR 3′-UTR. MiR-615 and epidermal growth factor receptor (EGFR) are associated with a number of disease processes and pathogenesis. However, little is known about the mechanisms of miR-615 and EGFR in human glioblastoma multiforme (GBM). Here, we found that down-regulation of miR-615 expression occurred in GBM tissues and cells, and was inversely correlated with overall survival, relapse-free survival, WHO grade as well as EGFR expression. We further determined that miR-615 functions as a tumor suppressor by inhibiting GBM cell proliferation, cell cycle, migration and invasion, and promoting cell apoptosis. In-vivo assay validated the inhibition effect of miR-615 on tumor growth and EGFR expression. Luciferase reporter assays demonstrated that miR-615 targeted the 3′-untranslated region (3′-UTR) of EGFR. Besides, over-expression of EGFR reversed the inhibition effects of miR-615, while silencing of EGFR aggravated these inhibition effects. In conclusions, we identified that miR-615 plays a tumor suppressor role in GBM cell proliferation, migration and invasion by targeting EGFR expression, and miR-615 may act as a novel biomarker for early diagnosis or therapeutic targets of GBM.

60 APPLIED LIFE SCIENCES↗

TGF beta inhibits expression of SP-A, SP-B, SP-C, but not SP-D in human alveolar type II cells

Highlights: • TGF beta inhibits expression of SP-A, SP-B, SP-C, Fatty acid synthase, and ABCA3. • TGF beta does not inhibit the expression of SP-D. • TGF beta does not induce EMT changes in highly differentiated type II cells. TGF beta is a multifunctional cytokine that regulates alveolar epithelial cells as well as immune cells and fibroblasts. TGF beta inhibits surfactant protein A, B and C expression in fetal human lung and can inhibit type II cell proliferation induced by FGF7 (KGF). However, little is known about direct effects of TGF beta on adult human type II cells. We cultured alveolar type II cells under air/liquid interface conditions to maintain their state of differentiation with or without TGF beta. TGF beta markedly decreased expression of SP-A, SP-B, SP-C, fatty acid synthase, and the phospholipid transporter ABCA3. However, TGF beta increased protein levels of SP-D with little change in mRNA levels, indicating that it is regulated independently from other components of surfactant. TGF beta is a negative regulator of both the protein and the phospholipid components of surfactant. TGF beta did not induce EMT changes in highly differentiated human type II cells. SP-D is an important host defense molecule and regulated independently from the other surfactant proteins. Taken together these data are the first report of the effect of TGF beta on highly differentiated adult human type II cells. The effects on the surfactant system are likely important in the development of fibrotic lung diseases.

60 APPLIED LIFE SCIENCES↗

Down-regulation of FOXR2 inhibits non-small cell lung cancer cell proliferation and invasion through the Wnt/β-catenin signaling pathway

Highlights: • Expression of FOXR2 is up-regulated in NSCLC tissues and cell lines. • Down-regulation of FOXR2 inhibits NSCLC cell proliferation and invasion in vitro. • Down-regulation of FOXR2 inhibits NSCLC cell growth and metastasis in vivo. • Down-regulation of FOXR2 inhibits the Wnt/β-catenin pathway in NSCLC cells. Forkhead box R2 (FOXR2), a new member of the FOX family, is an important player in a wide range of cellular processes such as proliferation, migration, differentiation and apoptosis. Recently, FOXR2 has been reported to be implicated in cancer development. However, the biological functions of FOXR2 in non-small cell lung cancer (NSCLC) remain unclear. In this study, we investigated the specific role of FOXR2 in NSCLC. The results showed that down-regulation of FOXR2 significantly inhibited NSCLC cell proliferation and invasion in vitro and suppressed NSCLC cell growth and metastasis in vivo. In addition, the decrease in FOXR2 expression markedly reduced the protein levels of β-catenin, cyclinD1 and c-Myc and hence inactivated the Wnt/β-catenin pathway in NSCLC cells. Taken together, we concluded that FOXR2 might be considered as a promising therapeutic target for NSCLC treatment.

60 APPLIED LIFE SCIENCES↗

Apixaban attenuates ischemia-induced myocardial fibrosis by inhibition of Gq/PKC signaling

Highlights: • Apixaban attenuates ischemia-induced myocardial fibrosis. • Ischemia-induced myocardial fibrosis shows higher thrombin level. • Thrombin induced collagen deposition in cardiac fibroblasts. • Thrombin induced collagen deposition by activation of Gq/PKC signaling. • Inhibition of Gq/PKC blunts ischemia-induced myocardial fibrosis. It was previously found that patients with symptom of myocardial dysfunction had increased levels of thrombin. Apixaban is one of the novel oral anticoagulant drugs widely used in clinic. As the inhibitor of FXa (prothrombin), it inhibits prothrombin conversion into thrombin leading to thrombin deficiency in vivo. However, the effects of apixaban on myocardial fibrosis were still unclear, and the concomitant molecular mechanisms remain to be investigated. Here, we showed that myocardial fibrosis-bearing mice induced by continuous myocardial ischemia (MI) had higher levels of thrombin. Orally administration of apixaban significantly abrogated fibrosis condition and thrombin levels. In vitro, thrombin induced collagen deposition in primary cardiac fibroblasts in a dose-dependent manner. Mechanistic experiments showed that thrombin induced collagen deposition by activation of the Par-1-coupled Gq/PKC signaling. Genetic ablation of Gq or pharmacological inhibition of PKC effectively blunted thrombin-induced collagen deposition in cardiac fibroblasts. Moreover, administration of PKC inhibitor or Gq antagonist obviously blocked MI-induced myocardial fibrosis in mice. To conclude, apixaban attenuates MI-induced myocardial fibrosis by inhibition of thrombin-dependent Par-1/Gq/PKC signaling axis.

60 APPLIED LIFE SCIENCES↗

MiR-130a-3p inhibits migration and invasion by regulating RAB5B in human breast cancer stem cell-like cells

Highlights: • We enriched breast cancer stem-like cells expressing CD44{sup +}CD24{sup -/low} from MCF-7 cells. • We found that miR-130a-3p is down-regulated in human breast cancer tissues and exosomes from circulating blood, and overexpression of miR-130a-3p in BCSCs inhibited cellular proliferation, migration, and invasion. • MiR-130a-3p inhibits migration and invasion by regulating RAB5B in human breast cancer stem cell-like cells. Breast cancer stem cells (BCSCs) constitute a subpopulation of tumor cells that express stem cell-associated markers and have a high capacity for tumor generation in vivo. MicroRNAs (miRNAs) are involved in tumorigenesis by regulating specific oncogenes and tumor suppressor genes, and their roles in BCSCs are becoming more apparent. We try to reveal the mechanism by which specific miRNA plays its function in BCSCs. Herein, we show that miR-130a-3p is down-regulated in human breast cancer tissues and exosomes from circulating blood. Overexpression of miR-130a-3p in BCSCs inhibited cellular proliferation, migration, and invasion, and silencing of miR-130a-3p had the opposite effects. We also confirmed that RAB5B is directly down-regulated by miR-130a-3p. Knockdown of RAB5B also inhibited cell proliferation, migration and invasion. Furthermore, we found that lower levels of exosome-derived miR-130a-3p are associated with lymph node metastasis and advanced TNM stage. Taken together, our results demonstrate that miR-130a-3p may act as a disease progression monitoring indicator and therapeutic target in breast cancer.

60 APPLIED LIFE SCIENCES↗

Inhibition of pancreatic cancer Panc1 cell migration by omeprazole is dependent on aryl hydrocarbon receptor activation of JNK

Highlights: • Omeprazole inhibition of Panc1 cell invasion is JNK- and AhR-dependent. • The non-genomic pathway is not due to AhR mutations. • Omeprazole-AhR-JNK modulates expression of invasion genes. Several aryl hydrocarbon receptor (AhR)-active pharmaceuticals were screened as inhibitors of pancreatic cancer cell invasion and identified two compounds, omeprazole, that inhibited invasion. Inhibition of highly invasive Panc1 cell invasion by omeprazole involves an AhR-dependent non-genomic pathway, and omeprazole-mediated inhibition of Panc1 cell invasion was dependent on Jun-N-terminal kinase (JNK) and mitogen-activated kinase kinase 7 (MKK7). The failure of omeprazole to induce nuclear translocation of the AhR was not due to overexpression of cytosolic AhR partner proteins Hsp90 or XAP2, and results of DNA sequencing show that the AhR expressed in Panc1 cells was not mutated. Results of RNAseq studies indicate that omeprazole induced an AhR-dependent downregulation of several pro-invasion factors including activated leukocyte cell adhesion molecule (ALCAM), long chain fatty acid CoA-synthase (CSL4), stathmin 3 (STMN3) and neuropillin 2 (NRP2), and the specific functions of these genes are currently being investigated.

60 APPLIED LIFE SCIENCES↗

Ataxin-3 promotes testicular cancer cell proliferation by inhibiting anti-oncogene PTEN

Highlights: • Ataxin-3 is up-regulated in TC tissues. • Ataxin-3 promoted TC cell proliferation. • Ataxin-3 inhibited the expression of PTEN in TC cells. • Ataxin-3 indirectly activated the AKT/mTOR signaling pathway in TC cells. Human Ataxin-3 protein was first identified as a transcript from patients with Machado-Joseph disease (MJD), also known as spinocerebellar ataxia type 3 (SCA3). Recent studies have demonstrated that Ataxin-3 is involved in gastric cancer and lung cancer. However, the role of Ataxin-3 in testicular cancer (TC) remains poorly understood. This study aims to explore the significance of Ataxin-3 expression in TC. Firstly, we investigated 53 paired TC and para-tumor tissues and found that Ataxin-3 was overexpressed in TC tissues, and this overexpression of Ataxin-3 was correlated with tumor stages. Functionally, Ataxin-3 overexpression promoted cell proliferation, and Ataxin-3 knockdown inhibited cell proliferation. In addition, up-regulation of Ataxin-3 inhibited the expression of PTEN and activated the AKT/mTOR pathway. Conversely, inhibition of Ataxin-3 suppressed the expression of p-AKT and p-mTOR, and increased the expression of p-4EBP1. These findings may provide a better understanding about the mechanism of TC and suggest that Ataxin-3 may be a potential prognostic biomarker and therapeutic target for TC.

60 APPLIED LIFE SCIENCES↗

Knockdown of ADAM17 inhibits cell proliferation and increases oxaliplatin sensitivity in HCT-8 colorectal cancer through EGFR-PI3K-AKT activation

Highlights: • High expression of ADAM17 is associated with oxaliplatin chemosensitivity. • Chronic oxaliplatin exposure established drug-resistant cell line HCT-8/L-OHP. • Down regulation of ADAM17 can enhance sensitivity and reverse resistance to oxaliplatin. • Downregulation of ADAM17 inhibit cell proliferation, induce apoptosis via EGFR/PI3K/AKT signaling pathway. We investigated the role of a disintegrin and metalloproteinase 17 (ADAM17) in chemo resistance, and to clarify the mechanism underlying reverse of L-OHP resistance by knockdown of ADAM17. CRC tissues with corresponding adjacent normal tissues were collected. The mRNA and protein expression of ADAM17 in tissues were detected by RT-qPCR, immunohistochemistry and Western blot. The prognostic impact of ADAM17 expression were then validated in TCGA database to confirm the results. Resistance to oxaliplatin was induced in HCT-8 (HCT-8/L-OHP) colorectal cancer cell line by exposing cell to increasing concentrations of L-OHP. MTT were used to evaluate the resistance to L-OHP. Subsequently, Knockdown of ADAM17 in HCT-8 and HCT-8/L-OHP cells to explore the mechanism through which ADAM17 shRNA reverses L-OHP resistance. Our result showed that ADAM17 was higher expression in the cancerous tissue and related to the chemosensitivity. Moreover, ADAM17 shRNA, AG1478 and LY294002 could inhibit cell proliferation, induce apoptosis and increase oxaliplatin sensitivity in HCT-8/L-OHP and parental colorectal cancer cell line, but nonsense shRNA did not show this effect. Western blot analysis further confirmed that EGFR/PI3K/AKT signaling pathway is involved in ADAM17 shRNA inhibiting proliferation and chemosensitivity of HCT-8/L-OHP and HCT-8 cells. The present study provides the evidence that downregulation of ADAM17 could increase the sensitivity to chemotherapy, inhibit cell proliferation, induce apoptosis, and reverse oxaliplatin resistance via suppression of the EGFR/PI3K/AKT signaling pathway in CRC.

60 APPLIED LIFE SCIENCES↗

PARP-1 inhibition attenuates cardiac fibrosis induced by myocardial infarction through regulating autophagy

Post-MI heart failure is characterized by structural remodeling, in which intramyocardial fibrosis takes a important part. Poly(ADP-ribose) polymerase 1 (PARP-1) is a extensive nuclear enzyme and plays a critical role in various diseases. It was shown that PARP-1 inhibition could alleviate heart failure and dowregulate autophagy, but whether PARP-1 regulates autophagy and thus impacts the activities of CFs remain unknown. We transfected cultured cardiac fibroblasts (CFs) with small interfere RNA-PARP-1 (siPARP-1) to downregulate PARP-1 and analyzed the ability of proliferation, migration, differentiation, and autophagy levels of CFs under different treatments using CCK8 assays, transwell migration assays, immunofluorescence assays detecting expression of α-SMA, western blot assays detecting autophagy-related proteins respectively. Furthermore, rat models of myocardial infarction (MI) were induced by ligation of left anterior descending coronary artery and PARP-1 inhibitor, 4-aminobenzamide (4-AB), was injected intraperitoneally after MI, followed by echocardiography detection, masson assays, immunohistochemistry assays detecting expression of α-SMA and western blot assays detecting autophagy-related proteins to investigate whether PARP-1 inhibition could regulate autophagy, alleviate cardiac fibrosis and improve cardiac function in vivo. In cultured CFs, siPARP-1 repressed TGF-β1-induced proliferation, migration, and differentiation through regulating autophagic levels. The in vitro results was verified by the in vivo study, indicating that PARP-1 inhibition partially decreased autophagy, abrogated cardiac fibrosis and significantly improved cardiac function post-MI. In conclusion, this work demonstrated the vital connection of PARP-1 and autophagy in the activation of CFs, and provided solid evidence supporting PARP-1 inhibition as a feasible strategy for the treatment of post-MI heart failure.

60 APPLIED LIFE SCIENCES↗

NKILA inhibition protects retinal pigment epithelium cells from hypoxia by facilitating NFκB activation

Highlights: • Hypoxia induces NKILA expression in retinal pigment epithelium (RPE) cells. • Hypoxia induces NKILA-IκBα association and NFκB activation in RPE cells. • NKILA shRNA inhibits hypoxia-induced RPE cell death. • NKILA overexpression potentiates hypoxia-induced RPE cell death. • miR-103 inhibition by hypoxia could be a primary reason of NKILA upregulation. Sustained retinal hypoxia causes injuries to retinal pigment epithelium (RPE) cells. We studied expression and potential functions of nuclear factor-κB (NFκB) Interacting LncRNA (NKILA) in hypoxia-treated RPE cells. Hypoxia induced NKILA expression, NKILA-IκBα association and NFκB activation in ARPE-19 cells and primary human RPE cells. shRNA-mediated knockdown of NKILA facilitated NFκB activation, inhibiting RPE cell death and apoptosis. Conversely, exogenous overexpression of NKILA blocked hypoxia-induced NFκB activation, thereby exacerbating RPE cell apoptosis. Further studies show that hypoxia downregulated microRNA-103 (miR-103), the anti-NKILA microRNA, in RPE cells. Transfection of miR-103 mimic blocked hypoxia-induced NKILA expression to significantly boost NFκB activation, protecting RPE cells from hypoxia. Collectively, we conclude that hypoxia-induced NKILA expression negatively regulates NFκB to promote RPE cell death. Conversely, NKILA inhibition protects RPE cells from hypoxia by facilitating NFκB activation.

60 APPLIED LIFE SCIENCES↗

Differential inhibition of mucin-type O-glycosylation (MTOG) induced by peracetyl N-thioglycolyl-d-galactosamine (Ac5GalNTGc) in myeloid cells

Highlights: • Each cell type expresses specific set of glycosyl transferases that result in unique pattern of mucin-type O-glycosylation. • MTOG of CD antigens is regulated depending on the specific cell type, developmental stage, and resting or activated state. • Metabolic incorporation of N-thioglycolyl-d-galactosamine results in cell-type dependent differential inhibition of MTOG. • Myeloid cells metabolize Ac{sub 5}GalNTGc with varying efficiency resulting in differential inhibition of MTOG on CD43. Investigations on the structure and functional roles of glycosylation – an intricate, complex, and dynamic post translational modification on proteins – in biological processes has been a challenging task. Glycan modifications vary depending on the specific cell type, its developmental stage, and resting or activated state. In the present study, we aim to understand the differences between the mucin-type O-glycosylation (MTOG) of two functionally divergent human cell lines, K562 (chronic myeloid leukemia) and U937 (histiocytic lymphoma), having myeloid origins. MTOG is initiated by the addition of N-acetyl-α-d-galactosamine (GalNAc) to Ser/Thr of glycoproteins. We exploited the metabolic glycan engineering (MGE) strategy using the peracetyl N-thioglycolyl-d-galactosamine (Ac{sub 5}GalNTGc), a synthetic GalNAc analogue, to engineer the glycoconjugates. Ac{sub 5}GalNTGc was metabolized and incorporated as N-thioglycolyl-d-galactosamine (GalNTGc) in cell surface glycoproteins in both the cell lines with varying degrees of efficiency. Notably, metabolic incorporation of GalNTGc resulted in differential inhibition of MTOG. It was observed that endogenous glycosylation machinery of K562 is relatively more stringent for selecting GalNTGc whereas U937 is flexible towards this selection. Additionally, we studied how the glycan modifications vary on a given CD antigen in these cell lines. Particularly, MTOG on CD43 was differentially inhibited in K562 and U937 as revealed by glycan-dependent and glycan-independent antibodies. It was observed that the effect of MGE on CD43 was similar to global effects on both cell lines. Consequences of MGE using GalNAc analogues depend on the expression and activity of various glycosyl transferases which determine global glycosylation on cell surface as well as on specific glycoproteins.

60 APPLIED LIFE SCIENCES↗

LINC01510 suppresses cell proliferation and invasion by inhibiting Wnt/β-catenin signaling in renal cell carcinoma

Highlights: • LINC01510 is down-regulated in renal cell carcinoma (RCC) and predicts poor prognosis. • LINC01510 inhibits RCC cell proliferation and growth by affecting cell cycle transition from G0/G1 to S stage. • LINC01510 inhibits the expression of MMP2, MMP7 and MMP9, and then decrease the ability of RCC cell invasion. • LINC01510 regulates cell proliferation and invasion through Wnt/β-catenin signaling. The role of long non-coding RNA in Renal cell carcinoma (RCC) tumorigenesis and progression remains largely unknown. Here, we found that LINC01510 functions as a tumor suppressor in RCC tumorigenesis. We screened TCGA database and then found that LINC01510 is significantly down-regulated in malignant RCC tissues, and the lower expression of LINC01510 predicts poor prognosis. Moreover, the down-regulated LINC01510 was further confirmed in our fresh tissues and cell lines. Biological functions assays shown that Ectopic expression of LINC01510 not only inhibits RCC cell proliferation both in vitro and in vivo, but also impairs cell invasion ability. Moreover, we found overexpression of LINC01510 inhibits the expression of CCND1 and CCNE1, as well as MMPs (MMP2, MMP7 and MMP9), and thus affecting RCC cell cycle and invasion. Meanwhile, Western blot assays revealed that the expression of β-catenin is regulated by LINC01510; overexpression of β-catenin could partly rescue the cell viability and invasion ability caused by ectopic expression of LINC01510. Taken together, we found that LINC01510 regulates cell proliferation and invasion by modulating Wnt/β-catenin signaling in RCC.

60 APPLIED LIFE SCIENCES↗

ODC1 inhibits the inflammatory response and ROS-induced apoptosis in macrophages

Highlights: • ODC1 is up-regulated by LPS, BCG and CCl4 stimulation. • ODC1 inhibits inflammatory responses in macrophages. • ODC1 mediates NF-κB and MAPK activation. • ODC1 inhibits ROS-induced macrophage apoptosis. Macrophage activation plays a critical role in the innate immune response. Ornithine decarboxylase (ODC1) metabolizes l-ornithine to polyamines and is the rate-limiting enzyme involved in the metabolism of polyamines, which are reportedly involved in cell differentiation, proliferation, and migration. However, the function of ODC1 in immune cells and especially in macrophages, as well as its underlying molecular mechanism, remains unclear. This study revealed the potential ODC1 function and mechanism associated with the lipopolysaccharide (LPS)-, Bacillus Calmette–Guerin (BCG)-, or carbon tetrachloride (CCl{sub 4})-induced inflammatory response in macrophages. We found significant upregulation of ODC1 in macrophages following LPS simulation and ODC1-specific suppression of proinflammatory cytokine secretion from macrophages upon stimulation with LPS, BCG and CCl{sub 4}, respectively, suggesting a role as a common control element of the inflammatory response. Western blotting for nuclear factor-κB and mitogen-activated protein kinases revealed significant inhibition of phosphorylation of multiple transcription factors following ODC1 overexpression in macrophages. Moreover, ODC1 inhibited reactive oxygen species-induced and caspase-dependent apoptosis highlighted by decreased caspase-3 and -9 expression following ODC1 upregulation. These findings indicated that ODC1 was involved in attenuating the inflammatory response upon stimulation of macrophages, making it a potential therapeutic target for inflammatory diseases.

60 APPLIED LIFE SCIENCES↗

Sulindac sulfone inhibits the mTORC1 pathway in colon cancer cells by directly targeting voltage-dependent anion channel 1 and 2

Highlights: • We identified VDAC1 and VDAC2 as novel binding proteins of sulindac sulfone. • Sulindac sulfone directly binds to both VDAC1 and VDAC2. • Sulindac sulfone may negatively regulate the function of VDAC. • Sulindac sulfone may inhibit the mTORC1 pathway through the inhibition of VDAC. Sulindac sulfone is a metabolite of sulindac, a non-steroidal anti-inflammatory drug (NSAID), without anti-inflammatory ability. However, sulindac sulfone has been reported to significantly reduce polyps in patients with colorectal adenomatous polyposis in clinical trials. Thus, sulindac sulfone is expected to be useful for the chemoprevention of neoplasia with few side effects related to anti-inflammatory ability. To date, the molecular targets of sulindac sulfone have not yet fully investigated. Therefore, in order to newly identify sulindac sulfone-binding proteins, we generated sulindac sulfone-fixed FG beads and purified sulindac sulfone-binding proteins from human colon cancer HT-29 cells. we identified mitochondrial outer membrane proteins voltage-dependent anion channel (VDAC) 1 and VDAC2 as novel molecular targets of sulindac sulfone, and sulindac sulfone directly bound to both VDAC1 and VDAC2. Double knockdown of VDAC1 and VDAC2 by siRNA inhibited growth and arrested the cell cycle at G1 phase in HT-29 cells. Depletion of VDAC1 and VDAC2 also inhibited the mTORC1 pathway with a reduction in cyclin D1. Interestingly, these effects were consistent with those of sulindac sulfone against human colon cancer cells, suggesting that sulindac sulfone negatively regulates the function of VDAC1 and VDAC2. In the present study, our data suggested that VDAC1 and VDAC2 are direct targets of sulindac sulfone which suppresses the mTORC1 pathway and induces G1 arrest.

60 APPLIED LIFE SCIENCES↗

Clotrimazole inhibits the Wnt/β-catenin pathway by activating two eIF2α kinases: The heme-regulated translational inhibitor and the double-stranded RNA-induced protein kinase

Highlights: • A zebrafish phenotypic screening repositioned antifungal azoles to Wnt inhibitors. • Clotrimazole reduces β-catenin levels in a proteasome-independent manner. • Clotrimazole inhibits Wnt/β-catenin signaling by activating eIF2α kinases. The Wnt/β-catenin signaling pathway controls cell proliferation and differentiation, and therefore, when this pathway is excessively activated, it causes tumorigenesis. Our chemical suppressor screening in zebrafish embryos identified antifungal azoles including clotrimazole, miconazole, and itraconazole, as Wnt/β-catenin signaling inhibitors. Here we show the mechanism underlying the Wnt/β-catenin pathway inhibition by antifungal azoles. Clotrimazole reduced β-catenin revels in a proteasome-independent fashion. By gene knockdown of two translational regulators, heme-regulated translational inhibitor and double-stranded RNA-induced protein kinase, we show that they mediate the clotrimazole-induced inhibition of the Wnt/β-catenin pathway. Thus, clotrimazole inhibits the Wnt/β-catenin pathway by decreasing β-catenin protein levels through translational regulation. Antifungal azoles represent genuine candidate compounds for anticancer drugs or chemopreventive agents that reduce adenomatous polyps.

60 APPLIED LIFE SCIENCES↗

Inhibition of cGAS-STING-TBK1 signaling pathway by DP96R of ASFV China 2018/1

Highlights: • DP96R is an inhibitor of cGAS-triggered type I IFNs induction and NF-κB activation. • DP96R inhibits the phosphorylation of TBK1 and TBK1-mediated antiviral responses. • The C-terminal domain of DP96R is responsible for its inhibitory effects. African swine fever virus (ASFV) is a highly pathogenic large DNA virus that causes African swine fever (ASF) in domestic pigs and European wild boars with mortality rate up to 100%. The DP96R gene of ASFV encodes one of the viral virulence factors, yet its action mechanism remains unknown. In this study, we report that DP96R of ASFV China 2018/1 strain subverts type I IFN production in cGAS sensing pathway. DP96R inhibited the cGAS/STING, and TBK1 but not IRF3-5D mediated IFN-β and ISRE promoters activation. Furthermore, DP96R selectively blocked the activation of NF-κB promoter induced by cGAS/STING, TBK1, and IKKβ, but not by overexpression of p65. Moreover, DP96R inhibited phosphorylation of TBK1 stimulated by cGAS/STING activation, and TBK1-induced antiviral response. Finally, truncated mutation analysis demonstrated that the region spanning amino acids 30 to 96 of DP96R was responsible for the inhibitory activity. To our knowledge, this is for the first time that DP96R of ASFV China 2018/1 is reported to negatively regulate type I IFN expression and NF-κB signaling by inhibiting both TBK1 and IKKβ, which plays an important role in virus immune evasion.

60 APPLIED LIFE SCIENCES↗

MicroRNA-98-5p ameliorates oxygen-glucose deprivation/reoxygenation (OGD/R)-induced neuronal injury by inhibiting Bach1 and promoting Nrf2/ARE signaling

Highlights: • miR-98 expression was altered in neurons with OGD/R treatment. • miR-98-5p inhibited OGD/R-induced apoptosis and ROS production. • miR-98-5p targets and regulates Bach1. • miR-98-5p regulates OGD/R-induced injury by targeting Bach1. • miR-98-5p regulates Nrf2/ARE signaling by targeting Bach1. MicroRNA-98-5p (miR-98-5p) is a stress-related microRNA (miRNA) that plays an important role in regulating cell survival, apoptosis, and oxidative stress in multiple cell types and diseases. However, little is known about the role of miR-98-5p in cerebral ischemia/reperfusion injury. In this study, we investigated the role and mechanism of miR-98-5p in regulating neuronal injury induced by oxygen-glucose deprivation/reoxygenation (OGD/R), an in vitro model of cerebral ischemia/reperfusion injury. We found that miR-98 expression was significantly altered in neurons in response to OGD/R treatment. Functional experiments showed that overexpression of miR-98-5p inhibited OGD/R-induced apoptosis and reactive oxygen species (ROS) production in neurons, whereas inhibition of miR-98-5p showed the opposite effect. Interestingly, bioinformatics analysis predicted that BTB and CNC homology 1 (Bach1) was a potential target gene of miR-98-5p, that was verified by dual-luciferase reporter assay. Moreover, overexpression of miR-98-5p inhibited Bach1 expression while suppression of miR-98-5p promoted Bach1 expression in neurons. Notably, miR-98-5p was shown to regulate the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) and the activity of the antioxidant response element (ARE). However, overexpression of Bach1 or silencing of Nrf2 significantly abolished the miR-98-5p-mediated neuroprotective effect. Overall, these results demonstrate that miR-98-5p ameliorates OGD/R-induced neuronal injury in vitro through targeting to promote activation of Nrf2/ARE signaling. Our study suggests that miR-98-5p may play a potential role in cerebral ischemia/reperfusion injury and represents a potential therapeutic target for neuroprotection.

60 APPLIED LIFE SCIENCES↗

Inhibition of Drp1 hyperactivation reduces neuropathology and behavioral deficits in zQ175 knock-in mouse model of Huntington's disease

Highlights: • Inhibition of Drp1 hyperactivation reduces behavioral deficits in zQ175 knock-in mice. • Inhibition of Drp1 hyperactivation attenuates striatal cell death in HD mice. • Suppression of Drp1 hyperactivation reduces white matter disorganization in HD mice. Mitochondrial dysfunction manifests in the pathogenesis of Huntington's disease (HD), a fatal and inherited neurodegenerative disease. Dynamin-related protein 1 (Drp1) is the primary component of mitochondrial fission and becomes hyperactivated in various models of HD. We previously reported that inhibition of Drp1 hyperactivation by P110, a rationally designed peptide inhibitor of Drp1-Fis1 interaction, is protective in the HD R6/2 mouse model, which expresses a fragment of mutant Huntingtin (mHtt). In this study, we expand our work to test the effect of P110 treatment in HD knock-in (zQ175 KI) mice that express full-length mtHtt and exhibit progressive disease symptoms, reminiscent of human HD. We find that subcutaneously sustained treatment with P110 reduces movement deficits of mice. Moreover, the treatment attenuates striatal neuronal loss, microglial hyperactivity and white matter disorganization in zQ175 KI mice. These findings provide an additional line of evidence that inhibition of Drp1 hyperactivation is sufficient to reduce HD-associated neuropathology and behavioral deficits. We propose that manipulation of Drp1 hyperactivation might be a useful strategy to develop therapeutics for treating HD.

60 APPLIED LIFE SCIENCES↗