• Journal of Microbiology and Biotechnology
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• 제33권1호
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• pp.114-122
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• 2023

A family of signal transduction pathways known as wingless type (Wnt) signaling pathways is essential to developmental processes like cell division and proliferation. Mutation in Wnt signaling results in a variety of diseases, including cancers of the breast, colon, and skin, metabolic disease, and neurodegenerative disease; thus, the Wnt signaling pathways have been attractive targets for disease treatment. However, the complicatedness and large involveness of the pathway often hampers pinpointing the specific targets of the metabolic process. In our current study, we investigated the differential metabolic regulation by the overexpression of the Wnt signaling pathway in a timely-resolved manner by applying high-throughput and un-targeted metabolite profiling. We have detected and annotated 321 metabolite peaks from a total of 36 human embryonic kidney (HEK) 293 cells using GC-TOF MS and LC-Orbitrap MS. The un-targeted metabolomic analysis identified the radical reprogramming of a range of central carbon/nitrogen metabolism pathways, including glycolysis, TCA cycle, and glutaminolysis, and fatty acid pathways. The investigation, combined with targeted mRNA profiles, elucidated an explicit understanding of activated fatty acid metabolism (β-oxidation and biosynthesis). The findings proposed detailed mechanistic biochemical dynamics in response to Wnt-driven metabolic changes, which may help design precise therapeutic targets for Wnt-related diseases.

• 대한의용생체공학회:의공학회지
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• 제41권6호
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• pp.235-246
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• 2020

This study aimed to evaluate the inhibitory effect of micro-current stimulation(MCS) on adipogenesis regarding with Wnt/β-catenin pathway using the ob/ob mouse and 3T3-L1 cell line. 6-week old ob/ob male mice were equally assigned to four groups: obese group(ob), obese with MCS groups(50 μA, 200 μA, and 400 μA). 6-week old C57BL/6J male mice were assigned to the control group(CON). We analyzed abdominal adipose tissue volume by using in vivo micro-CT and measured the body weight, feed intake, liver weight and triglycerides in serum. All the MCS groups showed that significantly reduced body weight and triglycerides in serum. In the case of liver weight and abdominal adipose tissue volume, the inhibitory effect of adipogenesis was shown in the 200 μA and 400 μA groups. To elucidate the anti-obesity effect of MCS, β-catenin, C/EBPα and FAS protein expressions were analyzed by western blotting. β-catenin expression was upregulated, C/EBPα and FAS expression were down-regulated in the relatively high-intensity groups(200 μA and 400 μA). Thus, the 200 μA and 400 μA for the intensity of MCS were chosen for cell experiments. In the 3T3-L1 cell line, Wnt/β-catenin pathway including Wnt10b, Wnt3a, β-catenin and Cyclin D1 was activated in all MCS groups. Accordingly, the expression level of C/EBPα was decreased during the differentiation and lipid droplet was significantly reduced in Oil red O staining results. These results suggest that the Wnt/β-catenin signaling might be activated by MCS with current intensities between 200-400 μA and it may lead to anti-obesity effects.

• Genomics & Informatics
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• 제19권1호
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• pp.5.1-5.11
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• 2021

Head and neck squamous cell carcinoma (HNSCC) is the most frequent type of head and neck cancer that usually arises from the mucosal surfaces of several organs including nasal cavity, paranasal sinuses, oral cavity, tongue, pharynx, and larynx. The Wnt signaling pathway is a crucial mechanism for cellular maintenance and development. It regulates cell cycle progression, apoptosis, proliferation, migration, and differentiation. Dysregulation of this pathway correlates with oncogenesis in various tissues including breast, colon, pancreatic as well as head and neck cancers. The present study aims to assess the gene alterations in the Wnt family of genes so as to derive an association with HNSCC. Computational approaches have been utilized for the identification of gene alterations in the Wnt family of genes. Several databases such as cBioportal, STRING, and UALCAN were used for the purpose. The frequency of alteration was high in case of Wnt family member 11 (5%). Gene amplification, deep deletions, missense and truncating mutations were observed in HNSCC patients. There was a marked difference in the gene expression profile of WNT11 between grades as well as normal samples. The survival probability measured using the Kaplan-Meier curve also presented with a significant difference among male and female subjects experiencing a low/medium level expression. The female patients showed less survival probability when compared to the male subjects. This provides the prognostic significance of the WNT11 gene in HNSCC. Taken together, the present study provides clues on the possible association of WNT11 gene alterations with HNSCC, which has to be further validated using experimental approaches.

• BMB Reports
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• 제49권7호
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• pp.357-358
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• 2016

Merlin, encoded by the NF2 gene, is a tumor suppressor that exerts its function via inhibiting mitogenic receptors at the plasma membrane. Although multiple mutations in Merlin have been identified in Neurofibromatosis type II (NF2) disease, its molecular mechanism is not fully understood. Here, we show that Merlin interacts with LRP6 and inhibits LRP6 phosphorylation, a critical step for the initiation of Wnt signaling. We found that treatment of Wnt3a caused phosphorylation of Merlin by PAK1, leading to detachment of Merlin from LRP6 and allowing the initiation of Wnt/β-catenin signaling. A higher level of β-catenin was found in tissues from NF2 patients. Enhanced proliferation and migration caused by knockdown of Merlin in glioblastoma cells were inhibited by suppression of β-catenin. Conclusively, these results suggest that sustained Wnt/β-catenin signaling activity induced by abrogation of Merlin-mediated inhibition of LRP6 phosphorylation might be a cause of NF2 disease.

• Molecules and Cells
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• 제40권1호
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• pp.45-53
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• 2017

Aberrant hypermethylation of Wnt antagonists has been observed in gastric cancer. A number of studies have focused on the hypermethylation of a single Wnt antagonist and its role in regulating the activation of signaling. However, how the Wnt antagonists interacted to regulate the signaling pathway has not been reported. In the present study, we systematically investigated the methylation of some Wnt antagonist genes (SFRP2, SFRP4, SFRP5, DKK1, DKK2, and APC) and their regulatory role in carcinogenesis. We found that aberrant promoter methylation of SFRP2, SFRP4, DKK1, and DKK2 was significantly increased in gastric cancer. Moreover, concurrent hypermethylation of SFRP2 and DKK2 was observed in gastric cancer and this was significantly associated with increased expression of ${\beta}-catenin$, indicating that the joint inactivation of these two genes promoted the activation of the Wnt signaling pathway. Further analysis using a multivariate Cox proportional hazards model showed that DKK2 methylation was an independent prognostic factor for poor overall survival, and the predictive value was markedly enhanced when the combined methylation status of SFRP2 and DKK2 was considered. In addition, the methylation level of SFRP4 and DKK2 was correlated with the patient's age and tumor differentiation, respectively. In conclusion, epigenetic silencing of Wnt antagonists was associated with gastric carcinogenesis, and concurrent hypermethylation of SFRP2 and DKK2 could be a potential marker for a prognosis of poor overall survival.

• East Asian mathematical journal
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• 제39권1호
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• pp.65-73
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• 2023

In this paper, we present an optimal control strategy to prevent the EMT process by downregulating the level of overexpressed β-catenin in the cytoplasm. To do this, we propose a mathematical model that expresses relationship between the Wnt signaling pathway and TGF-β in cancer cells. We also define an optimal control problem considering the side effects that occur simultaneously with the method for controlling the concentration of β-catenin. Finally numerical simulations show that treatment effect is quantitatively changes depending on the concentration of core proteins of the Wnt signaling pathway.

• 통합자연과학논문집
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• 제12권4호
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• pp.133-141
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• 2019

Mesenchymal stem cells (MSCs) are known to differentiate into multiple lineages, making neurogenic differentiation an important target in the clinical field. In the present study, we induced the neurogenic differentiation of cells using histone deacetylase (HDAC) inhibitors and studied their mechanisms for further differentiation in vitro. We treated cells with the HDAC inhibitors, MS-275 and NaB; and found that the cells had neuron-like features such as distinct bipolar or multipolar morphologies with branched processes. The mRNA expressions encoding for NEFL, MAP2, TUJ1, OLIG2, and SYT was significantly increased following HDAC inhibitors treatment compared to without HDAC inhibitors; high protein levels of MAP2 and Tuj1 were detected by immunofluorescence staining. We examined the mechanisms of differentiation and found that the Wnt signaling pathway and downstream mitogen-activate protein kinase were involved in neurogenic differentiation of MSCs. Importantly, Wnt4, Wnt5a/b, and Wnt11 protein levels were highly increased after treatment with NaB; signals were activated through the regulation of Dvl2 and Dvl3. Interestingly, NaB treatment increased the levels of JNK and upregulated JNK phosphorylation. After MS-275 treatment, Wnt protein levels were decreased and GSK-3β was phosphorylated. In this cell, HDAC inhibitors controlled the non-canonical Wnt expression by activating JNK phosphorylation and the canonical Wnt signaling by targeting GSK-3β.

• Molecules and Cells
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• 제41권9호
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• pp.830-841
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• 2018

Recent studies have indicated that microRNAs (miRNAs) play an important role in hepatocellular carcinoma (HCC) progression. In this study, we showed that miR-766-3p was decreased in approximately 72% of HCC tissues and cell lines, and its low expression level was significantly correlated with tumour size, TNM stage, metastasis, and poor prognosis in HCC. Ectopic miR-766-3p expression inhibited HCC cell proliferation, colony formation, migration and invasion. In addition, we showed that miR-766-3p repressed Wnt3a expression. A luciferase reporter assay revealed that Wnt3a was a direct target of miR-766-3p, and an inverse correlation between miR-766-3p and Wnt3a expression was observed. Moreover, Wnt3a up-regulation reversed the effects of miR766-3p on HCC progression. In addition, our study showed that miR-766-3p up-regulation decreased the nuclear ${\beta}-catenin$ level and expression of Wnt targets (TCF1 and Survivin) and reduced the level of MAP protein regulator of cytokinesis 1 (PRC1). However, these effects of miR-766-3p were reversed by Wnt3a up-regulation. In addition, PRC1 upregulation increased the nuclear ${\beta}-catenin$ level and protein expression of TCF1 and Survivin. iCRT3, which disrupts the ${\beta}-catenin-TCF4$ interaction, repressed the TCF1, Survivin and PRC1 protein levels. Taken together, our results suggest that miR-766-3p down-regulation promotes HCC cell progression, probably by targeting the Wnt3a/PRC1 pathway, and miR-766-3p may serve as a potential therapeutic target in HCC.

• 생명과학회지
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• 제28권9호
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• pp.1016-1021
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• 2018

C. difficile 톡신A에 의한 대장상피세포 자살과정은 위막성대장염(Pseudomembranous colitis)의 주요 원인으로 고려되고 있다. 톡신A는 활성산소 를 증가시켜 세포자살 신호를 유도한다. 또한 톡신A는 미세섬유나 미세소관과 같은 세포골격계 형성을 저해함으로써 자살을 유도한다고 알려져 있다. 하지만 톡신A가 야기하는 소화기 상피세포 자살경로는 아직 불분명하다. 본 연구에서는 소화관 상피세포의 성장과 분화 그리고 기능에 중요하다고 알려져 온 Wnt 신호경로에 대한 톡신A의 영향을 확인해보았다. 이를 위해 비암화-인간대장세포주(NCM460)에 톡신A를 처치하고 Wnt 신호 분자들의 변화를 추적하였다. 또한 톡신A를 주입한 생쥐의 회장 상피세포 속 Wnt 신호경로 변화도 평가하였다. 인간 대장상피세포에서 톡신A는 Wnt 경로의 핵심 신호분자인 ${\beta}$-catenin 단백질의 양을 빠르게 감소시켰다. 이 현상은 생쥐 회장 상피세포에서도 동일하게 확인되었다. 연구자 등은 톡신A가 $GSK3{\beta}$ 활성형 인산화(Thr390)를 증가시킴도 확인하였다. 이는 톡신A가 $GSK3{\beta}$의 활성을 높여서 ${\beta}$-catenin의 인산화시키고 이를 통해 단백질 분해 과정이 촉진되었음을 보여준다. 이 결과들을 종합하면, 톡신A에 의한 소화관 상피세포 자살과정이 상피세포의 성장과 자살을 조절하는 Wnt 신호경로 차단과 밀접하게 연관되어 있음을 보여준다.

• The Korean Journal of Physiology and Pharmacology
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• 제28권1호
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• pp.21-30
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• 2024

The challenging clinical outcomes associated with advanced cervical cancer underscore the need for a novel therapeutic approach. Monensin, a polyether antibiotic, has recently emerged as a promising candidate with anti-cancer properties. In line with these ongoing efforts, our study presents compelling evidence of monensin's potent efficacy in cervical cancer. Monensin exerts a pronounced inhibitory impact on proliferation and anchorage-independent growth. Additionally, monensin significantly inhibited cervical cancer growth in vivo without causing any discernible toxicity in mice. Mechanism studies show that monensin's anti-cervical cancer activity can be attributed to its capacity to inhibit the Wnt/β-catenin pathway, rather than inducing oxidative stress. Monensin effectively reduces both the levels and activity of β-catenin, and we identify Akt, rather than CK1, as the key player involved in monensin-mediated Wnt/β-catenin inhibition. Rescue studies using Wnt activator and β-catenin-overexpressing cells confirmed that β-catenin inhibition is the mechanism of monensin's action. As expected, cervical cancer cells exhibiting heightened Wnt/β-catenin activity display increased sensitivity to monensin treatment. In conclusion, our findings provide pre-clinical evidence that supports further exploration of monensin's potential for repurposing in cervical cancer therapy, particularly for patients exhibiting aberrant Wnt/β-catenin activation.