KSHV vPK inhibits Wnt signaling via preventing interactions between β-catenin and TCF4

Cha, Seho,Kang, Myung-Suk,Seo, Taegun Elsevier 2018 Biochemical and biophysical research communication Vol.497 No.1

<P><B>Abstract</B></P> <P>Viral factors interact with host cellular proteins, leading to dysregulation of signaling pathways. The Wnt pathway is known to participate in embryonic development and oncogenesis under dysregulation conditions. A downstream factor of the Wnt signaling pathway, β-catenin, activates T-cell factor (TCF)-dependent transcription, which contributes to cell proliferation and tumorigenesis. In this study, we demonstrated that viral protein kinase (vPK) encoded by Kaposi's sarcoma-associated herpesvirus inhibits the Wnt signaling pathway without affecting nuclear localization and expression of β-catenin. Coimmunoprecipitation and chromatin immunoprecipitation assays revealed that vPK interacts with β-catenin, reducing the binding affinity on TCF binding regions as well as interactions of β-catenin with TCF4. Overexpression of vPK led to reduced mRNA expression of cyclin D1, a well-known transcriptional product of Wnt signaling, suggesting that vPK effectively regulates the host signaling pathway through direct interactions with cellular proteins.</P> <P><B>Highlights</B></P> <P> <UL> <LI> vPK decreases LiCl-mediated activation of TCF-dependent transcription. </LI> <LI> vPK does not disrupt relocalization of β-catenin from the cytosol to nucleus. </LI> <LI> vPK disrupts interactions of β-catenin with the TCF4-binding region. </LI> <LI> vPK inhibits Wnt signaling via interactions with β-catenin. </LI> </UL> </P>

miR-458b-5p regulates ovarian granulosa cells proliferation through Wnt/β‐catenin signaling pathway by targeting catenin beta-1

Wang Wenwen,Teng Jun,Han Xu,Zhang Shen,Zhang Qin,Tang Hui 아세아·태평양축산학회 2021 Animal Bioscience Vol.34 No.6

Objective: Ovarian follicular development, which dependent on the proliferation and differentiation of granulosa cells (GCs), is a complex biological process in which miRNA plays an important role. Our previous study showed that miR-458b-5p is associated with ovarian follicular development in chicken. The detailed function and molecular mechanism of miR-458b-5p in GCs is unclear. Methods: The luciferase reporter assay was used to verify the targeting relationship between miR-458b-5p and catenin beta-1 (CTNNB1), which is an important transcriptional regulatory factor of the Wnt/β-catenin pathway. The cell counting kit-8 (CCK-8) assay, flow cytometry with propidium iodide (PI) and annexin V-fluorescein isothiocyanate (FITC) labeling were applied to explore the effect of miR-458b-5p on proliferation, cell cycle and apoptosis of chicken GCs. Quantitative real-time polymerase chain reaction and Western blot were used to detect the mRNA and protein expression levels. Results: We demonstrated that the expression of miR-458b-5p and CTNNB1 showed the opposite relationship in GCs and theca cells of hierarchical follicles. The luciferase reporter assay confirmed that CTNNB1 is the direct target of miR-458b-5p. Using CCK-8 assay and flow cytometry with PI and Annexin V-FITC labeling, we observed that transfection with the miR-458b-5p mimics significantly reduced proliferation and has no effects on apoptosis of chicken GCs. In addition, miR-458b-5p decreased the mRNA and protein expression of CD44 molecule and matrix metallopeptidase 7, which are the downstream effectors of CTNNB1 in Wnt/β-Catenin pathway and play functional roles in cell proliferation. Conclusion: Taken together, the data indicate that miR-458b-5p regulates ovarian GCs proliferation through Wnt/β-catenin signaling pathway by targeting CTNNB1, suggesting that miR-458b-5p and its target gene CTNNB1 may potentially play a role in chicken ovarian follicular development. Objective: Ovarian follicular development, which dependent on the proliferation and differentiation of granulosa cells (GCs), is a complex biological process in which miRNA plays an important role. Our previous study showed that miR-458b-5p is associated with ovarian follicular development in chicken. The detailed function and molecular mechanism of miR-458b-5p in GCs is unclear.Methods: The luciferase reporter assay was used to verify the targeting relationship between miR-458b-5p and catenin beta-1 (<i><i>CTNNB1</i></i>), which is an important transcriptional regulatory factor of the Wnt/β-catenin pathway. The cell counting kit-8 (CCK-8) assay, flow cytometry with propidium iodide (PI) and annexin V-fluorescein isothiocyanate (FITC) labeling were applied to explore the effect of miR-458b-5p on proliferation, cell cycle and apoptosis of chicken GCs. Quantitative real-time polymerase chain reaction and Western blot were used to detect the mRNA and protein expression levels.Results: We demonstrated that the expression of miR-458b-5p and <i>CTNNB1</i> showed the opposite relationship in GCs and theca cells of hierarchical follicles. The luciferase reporter assay confirmed that <i>CTNNB1</i> is the direct target of miR-458b-5p. Using CCK-8 assay and flow cytometry with PI and Annexin V-FITC labeling, we observed that transfection with the miR-458b-5p mimics significantly reduced proliferation and has no effects on apoptosis of chicken GCs. In addition, miR-458b-5p decreased the mRNA and protein expression of CD44 molecule and matrix metallopeptidase 7, which are the downstream effectors of <i>CTNNB1</i> in Wnt/β-Catenin pathway and play functional roles in cell proliferation.Conclusion: Taken together, the data indicate that miR-458b-5p regulates ovarian GCs proliferation through Wnt/β-catenin signaling pathway by targeting <i>CTNNB1</i>, suggesting that miR-458b-5p and its target gene <i>CTNNB1</i> may potentially play a role in chicken ovarian follicular development.

SDC4 Gene Silencing Favors Human Papillary Thyroid Carcinoma Cell Apoptosis and Inhibits Epithelial Mesenchymal Transition via Wnt/β-Catenin Pathway

Liang-Liang Chen,Ge-Xin Gao,Fei-Xia Shen,Xiong Chen,Xiao-Hua Gong,Wen-Jun Wu 한국분자세포생물학회 2018 Molecules and cells Vol.41 No.9

As the most common type of endocrine malignancy, papillary thyroid cancer (PTC) accounts for 85-90% of all thyroid cancers. In this study, we presented the hypothesis that SDC4 gene silencing could effectively attenuate epithelial mesenchymal transition (EMT), and promote cell apoptosis via the Wnt/β-catenin signaling pathway in human PTC cells. Bioinformatics methods were employed to screen the determined differential expression levels of SDC4 in PTC and adjacent normal samples. PTC tissues and adjacent normal tissues were prepared and their respective levels of SDC4 protein positive expression, in addition to the mRNA and protein levels of SDC4, Wnt/β-catenin signaling pathway, EMT and apoptosis related genes were all detected accordingly. Flow cytometry was applied in order to detect cell cycle entry and apoptosis. Finally, analyses of PTC migration and invasion abilities were assessed by using a Transwell assay and scratch test. In PTC tissues, activated Wnt/β-catenin signaling pathway, increased EMT and repressed cell apoptosis were determined. Moreover, the PTC K1 and TPC-1 cell lines exhibiting the highest SDC4 expression were selected for further experiments. In vitro experiments revealed that SDC4 gene silencing could suppress cell migration, invasion and EMT, while acting to promote the apoptosis of PTC cells by inhibiting the activation of the Wnt/β-catenin signaling pathway. Besides, si-β-catenin was observed to inhibit the promotion of PTC cell migration and invasion caused by SDC4 overexpression. Our study revealed that SDC4 gene silencing represses EMT, and enhances cell apoptosis by suppressing the activation of the Wnt/β-catenin signaling pathway in human PTC.

P140 Light-emitting diode light regulates the proliferation of human outer root sheath cells via Wnt/β -catenin and ERK pathway

( Jung Eun Kim ),( Young Jun Woo ),( Kwan Ho Jeong ),( Hoon Kang ) 대한피부과학회 2016 대한피부과학회 학술발표대회집 Vol.68 No.2

<div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div><div style="display:none">fiogf49gjkf0d</div> Background: The hair growth promoting effects of low level light therapy (LLLT) have been reported. But the mechanisms on human outer root sheath cells (hORSCs) have been rarely studied. Objectives: We aimed to determine the effect of various wavelenths of light-emitting diode (LED) irradiation on hORSCs. Methods: The effects of the LED irradiation on cell proliferation, migration ability of hORSCs and the expression of molecules related to Wnt/β-catenin signaling and ERK pathway, hair stem cell markers, various growth factors and cytokines were examined. The effect of the LED irradiated hORSCs on cell proliferation of human dermal papilla cells (hDPCs) was also examined. Results: Low level irradiation of LED light variably promoted hORSC proliferation and suppressed cell apoptosis. LED irradiation induced Wnt5a, Axin2, Lef1 mRNA expression and β-catenin protein expression in hORSCs. Phosphorylation of ERK, c-Jun and p38 was also observed and PD98059 treatment before irradiation reduced the phosphorylation of ERK and c-Jun. The red light-treated ORSCs showed substantial upregulation of IL-6, IL-8, IGF-1, TGF-β1, and VEGF mRNA expressions. 660nm and 830nm of light irradiation to hORSC accelerated the migration in vitro. LED-irradiated hORSCs increased the proliferation of hDPCs when they are co-cultured. Conclusion: The growth promoting effect of LED on hORSC appears to be associated with the direct stimulation of the Wnt5a/β-catenin signaling and ERK signaling pathway.

Non-Polar Myxococcus fulvus KYC4048 Metabolites Exert Anti-Proliferative Effects via Inhibition of Wnt/β-Catenin Signaling in MCF-7 Breast Cancer Cells

( Juha Park ),( Hee-jin Yoo ),( Ah-ran Yu ),( Hye Ok Kim ),( Sang Cheol Park ),( Young Pyo Jang ),( Chayul Lee ),( Wonchae Choe ),( Sung Soo Kim ),( Insug Kang ),( Kyung-sik Yoon ) 한국미생물생명공학회(구 한국산업미생물학회) 2021 Journal of microbiology and biotechnology Vol.31 No.4

The Wnt/β-catenin signaling pathway is involved in breast cancer and Myxococcus fulvus KYC4048 is a myxobacterial strain that can produce a variety of bioactive secondary metabolites. Although a previous study revealed that KYC4048 metabolites exhibit anti-proliferative effects on breast cancer, the biochemical mechanism involved in their effects remains unclear. In the present study, KYC4048 metabolites were separated into polar and non-polar (ethyl acetate and n-hexane) fractions via liquid-liquid extraction. The effects of these polar and non-polar KYC4048 metabolites on the viability of breast cancer cells were then determined by MTT assay. Expression levels of Wnt/ β-catenin pathway proteins were determined by Western blot analysis. Cell cycle and apoptosis were measured via fluorescence-activated cell sorting (FACS). The results revealed that non-polar KYC4048 metabolites induced cell death of breast cancer cells and decreased expression levels of WNT2B, β-catenin, and Wnt target genes (c-Myc and cyclin D1). Moreover, the n-hexane fraction of non-polar KYC4048 metabolites was found most effective in inducing apoptosis, necrosis, and cell cycle arrest, leading us to conclude that it can induce apoptosis of breast cancer cells through the Wnt/β-catenin pathway. These findings provide evidence that the n-hexane fraction of non-polar KYC4048 metabolites can be developed as a potential therapeutic agent for breast cancer via inhibition of the Wnt/β-catenin pathway.

Elevated expression of Axin2 in intestinal metaplasia and gastric cancers

이동휘,정인호,장보근 대한병리학회 2023 Journal of Pathology and Translational Medicine Vol.57 No.6

Background: The Wnt signaling pathway regulates crucial cellular processes, including stem cell development and tissue repair. Dysregulation of this pathway, particularly β-catenin stabilization, is linked to colorectal carcinoma and other tumors. Axin2, a critical component in the pathway, plays a role in β-catenin regulation. This study examines Axin2 expression in normal gastric mucosa and various gastric pathologies. Methods: Formalin-fixed and paraffin-embedded tissue samples from normal stomach, gastritis, intestinal metaplasia (IM), and gastric carcinoma were collected. Axin2 and β-catenin expression were evaluated using RNA in situ hybridization and immunohistochemistry, respectively. Histo-scores (H-scores) were calculated to quantify expression levels of Axin2. Associations between Axin2 expression and clinicopathological variables were examined. Results: Axin2 expression was examined in normal stomach, gastritis, and IM tissues. Axin2 expression was mainly observed in the surface and isthmus areas in the normal stomach and gastritis, whereas Axin2 expression was markedly higher at the bases of IM. Axin2 H-scores were significantly elevated in IM (mean ± standard deviation [SD], 87.0 ± 38.9) compared to normal (mean ± SD, 18.0 ± 4.5) and gastritis tissues (mean ± SD, 33.0 ± 18.6). In total, 30% of gastric carcinomas showed higher Axin2 expression. Axin2 expression did not have significant associations with age, sex, Lauren classification, histological differentiation, invasion depth, and lymph node metastasis. However, a strong positive correlation was observed between Axin2 and nuclear β-catenin in gastric carcinomas (p < .001). Conclusions: Axin2 expression was significantly increased in IM compared to normal and gastritis cases. In addition, Axin2 showed a strong positive association with nuclear β-catenin expression in gastric carcinomas, demonstrating a close relationship with abnormal Wnt/β-catenin signaling pathway.

Various Wavelengths of Light-Emitting Diode Light Regulate the Proliferation of Human Dermal Papilla Cells and Hair Follicles via Wnt/β-Catenin and the Extracellular Signal-Regulated Kinase Pathways

( Hong Jin Joo ),( Kwan Ho Jeong ),( Jung Eun Kim ),( Hoon Kang ) 대한피부과학회 2017 Annals of Dermatology Vol.29 No.6

Background: The human dermal papilla cells (hDPCs) play an important role in regulation of hair cycling and growth. Objective: The aim of this study was to investigate the effect of different wavelengths of light-emitting diode (LED) irradiation on the proliferation of cultured hDPCs and on the growth of human hair follicles (HFs) in vitro. Methods: We examined the effect of LED irradiation on Wnt/β-catenin signaling and mitogen-activated protein kinase (MAPK) pathways in hDPCs. Anagen HFs were cultured with LED irradiation and elongation of each hair shaft was measured. Results: The most potent wavelength in promoting the hDPC proliferation is 660 nm and 830 nm promoted hDPC proliferation to a lesser extent than 660 nm. Various wavelengths significantly increased β-catenin, Axin2, Wnt3a, Wnt5a and Wnt10b mRNA expression. LED irradiation significantly increased β-catenin and cyclin D expression, and the phosphorylation of MAPK and extracellular signal-regulated kinase (ERK). HFs irradiated with 415 nm and 660 nm grew longer than control. Conclusion: Our result suggests that LED has a potential to stimulate hDPC proliferation via the activation of Wnt/β-catenin signaling and ERK pathway. To our best knowledge, this is the first report which investigated that the effect of various wavelengths of LED on hDPC proliferation and the underlying mechanisms. (Ann Dermatol 29(6) 747∼754, 2017)

Anti-Proliferative Activity of Nodosin, a Diterpenoid from Isodon serra, via Regulation of Wnt/β-Catenin Signaling Pathways in Human Colon Cancer Cells

( Eun Seo Bae ),( Young-mi Kim ),( Dong-hwa Kim ),( Woong Sub Byun ),( Hyen Joo Park ),( Young-won Chin ),( Sang Kook Lee ) 한국응용약물학회 2020 Biomolecules & Therapeutics(구 응용약물학회지) Vol.28 No.5

Colorectal cancer (CRC) is one of the most malignant type of cancers and its incidence is steadily increasing, due to life style factors that include western diet. Abnormal activation of canonical Wnt/β-catenin signaling pathway plays an important role in colorectal carcinogenesis. Therefore, targeting Wnt/β-catenin signaling has been considered a crucial strategy in the discovery of small molecules for CRC. In the present study, we found that Nodosin, an ent-kaurene diterpenoid isolated from Isodon serra, effectively inhibits the proliferation of human colon cancer HCT116 cells. Mechanistically, Nodosin effectively inhibited the overactivated transcriptional activity of β-catenin/T-cell factor (TCF) determined by Wnt/β-catenin reporter gene assay in HEK293 and HCT116 cells. The expression of Wnt/β-catenin target genes such as Axin2, cyclin D1, and survivin were also suppressed by Nodosin in HCT116 cells. Further study revealed that a longer exposure of Nodosin induced the G₂/M phase cell cycle arrest and subsequently apoptosis in HCT116 cells. These findings suggest that the anti-proliferative activity of Nodosin in colorectal cancer cells might in part be associated with the regulation of Wnt/β-catenin signaling pathway.

Circular RNAs Regulate Cancer Onset and Progression via Wnt/β-Catenin Signaling Pathway

Yun-Feng Li,Jian Zhang,Lei Yu 연세대학교의과대학 2019 Yonsei medical journal Vol.60 No.12

Cancer remains to be a major challenge for public health providers, and is the second leading cause of death worldwide. Therefore,it is imperative to explore the mechanisms underlying cancer initiation and development, and design novel diagnostics andtherapeutics. Circular RNAs (circRNAs), which exhibit a covalently closed loop structure, are involved in a variety of diseases, includingcancer. The aberrant expression of circRNAs contributes to the initiation and development of various cancers by disruptingthe interplay of specific signaling pathways, including the Wnt/β-catenin pathway, which controls a plethora of cellular processesthat drive cancer development. The interactions between circRNAs (specifically expressed in different cancer tissues) andWnt/β-catenin signaling pathway presents potential diagnostic biomarkers and novel therapeutic targets. In this review, we havesummarized research discoveries on the functions of Wnt/β-catenin pathway-related circRNAs in the modulation of oncogenesisand progression of different types of cancer. We anticipate that our findings will contribute to the improvement or developmentof circRNAs-based strategies for cancer treatment.

Electroacupuncture Promotes Neural Proliferation in Hippocampus of Perimenopausal Depression Rats via Wnt/β-Catenin Signaling Pathway

Qin Jing,Lu Ren,Xue Deng,Nan Zhang,Martin Fu,Ge Wang,Xi-Rong Jiang,Shu-Ru Lin,Cai-Rong Ming 사단법인약침학회 2020 Journal of Acupuncture & Meridian Studies Vol.13 No.3

Background and Objective: Perimenopausal depression is caused by the impaired function of the ovarium before menopause and with a series of symptoms. Electroacupuncture (EA) therapy has been demonstrated to improve clinically depression. However, the mechanism underlying its therapeutic activity remains unknown. This study aimed to investigat the effects of EA treatment on the hippocampal neural proliferation through Wnt signaling pathway. Methods: Chronic unpredictable mild stress (CUMS) combined with bilateral ovariectomy (OVX) were used to establish a rat model of perimenopausal depression. The open field test (OFT) and sucrose preference test (SPT) were used to assess depression-like behaviors in rats. ELISAs were used to measure estrogen (E2), luteinizing hormone (LH) and gonadotropin-releasing hormone (GnRH) levels in the serum. RT-PCR and Western blot assay were utilized for measuring the mRNA expressions and protein expressions of GSK-3β/β-catenin. Results: Four-week EA treatment at three points including “Shenshu” (BL23), “Baihui” (GV20) and “Sanyinjiao” (SP6) simultaneously ameliorated depression-like behaviors in rats with CUMS and OVX, whereas rescued the decreased serum level of E2 and prevented the increased serum levels of GnRH and LH. EA treatment ameliorated CUMS and OVX-induced alterations of glycogen synthase kinase-3b (GSK-3β) and β-catenin mRNA levels, β-catenin and phosphorylated β-catenin (p-β-catenin) protein levels. Conclusions: The results showed that EA treatment promoted hippocampal neural proliferation in perimenopausal depression rats via activating the Wnt/b-catenin signaling pathway, indicating that EA may represent an efficacious therapy for perimenopausal depression.