Suppression Of Wnt Signaling Pathway Research Articles

SMALL-MOLECULE TELOMERE INHIBITOR ATTENUATES THE LEUKEMIA-INITIATION POTENTIAL IN DRUG-RESISTANT LEUKEMIA CELLS

The drug resistance is the most often obstacle in leukemia treatment and failing to eradicating the quiescent leukemia stem cell (LSCs) is believed as the main reason of relapse. Numerous studies have indicated that abnormal maintenance of telomere is related to the sensitivity of cancer cells to therapy. Based on these facts, we designed a lot of compounds and conducted large scale screening. In this study, we identified a small molecule compound IX, an imatinib derivative, as a potential therapeutic candidate of drug-resistant leukemia treatment. IX can eradicate drug-resistant leukemia cells or LSCs, while has little effect on the normal progenitor cells in vitro. We also showed that IX can relive the tumor burden of CML xenograft mice model and prolong the life span of them. Meanwhile, it can inhibit the abnormal elongation of telomere not only by effectively decreases the telomerase activity of drug-resistant leukemia cells, but also by affecting ALT mechanism. Besides, the suppression of Wnt signaling pathway by IX may plays an important role in eradicating drug-resistant cells. Taken together, our findings suggest that IX could be a novel therapeutic agent for the treatment of refractory leukemia.

OxLDL inhibits differentiation of mesenchymal stem cells into osteoblasts via the CD36 mediated suppression of Wnt signaling pathway.

Bone abnormalities as a consequence of osteoblast deregulation are associated with several diseases such as diabetes and chronic kidney disease. Important role for oxidized low density lipoproteins (oxLDL) in the pathophysiology of bone disorders has been reported. However, little is known about the effects and mechanisms of oxLDL on the process of osteoblastogenesis in human mesenchymal stem cells (MSCs). We show that oxLDL concentrations of ~ 10-25µg protein (0.43-1.0µM MDA/mg protein) inhibited the differentiation of MSCs to osteoblasts. We demonstrate that the underlying mechanism entails the suppression of the Wnt signaling through the down-regulation of β-catenin. Further, we show the association of scavenger receptor CD36 with the receptors LRP5/6 and Frizzled in mediating the oxLDL effects on the differentiation of MSCs to pre-osteoblasts. Inhibiting CD36 restored osteoblasts differentiation in the presence of oxLDL. Our findings suggest that oxLDL interferes with the canonical Wnt signaling pathway in a CD36 dependent manner leading to an inhibition of osteoblastogenesis.

LGR5 regulates gastric adenocarcinoma cell proliferation and invasion via activating Wnt signaling pathway

LGR5 plays a critical role in tissue development and the maintenance of adult stem cells in gastrointestinal tract. However, the oncogenic role of LGR5 in the development of gastric adenocarcinoma remains elusive. Here, we show that LGR5 promotes gastric adenocarcinoma cell proliferation and metastasis. We find that knock down of LGR5 or suppression of Wnt signaling pathway by inhibitor C59 arrests gastric adenocarcinoma cell proliferation and invasion. Moreover, treatment of Wnt3a, the activator of Wnt signaling pathway, partially recovers the proliferation defect observed in LGR5 knockdown gastric adenocarcinoma cells. Moreover, LGR5 facilitates β-catenin nuclear accumulation, a surrogate marker of the activation of Wnt signaling pathway. In addition, C59 treatment suppresses transcription of Axin2 and TCF1, both of which are the target genes of β-catenin in gastric adenocarcinoma cells. Gastric adenocarcinoma cells with overexpressed LGR5 form a large quantity of visible actin filaments and pseudopods, suggesting that LGR5 significantly enhances the ability of cell movement, which might capacitate gastric adenocarcinoma cells with enhanced LGR5 expression to gain invasive and migratory properties. Taken together, our results show that LGR5 contributes to cell proliferation and invasion through the activation of Wnt/β-catenin-signaling pathway in gastric adenocarcinoma cells.

Porcine Circovirus-Like Virus P1 Inhibits Wnt Signaling Pathway in Vivo and in Vitro.

Porcine circovirus-like virus P1 is an important pathogen of the current pig industry, the infection mechanism is not entirely clear. Wnt signaling pathway plays an important role in the growth of young animals and infection of some viruses. This study was designed to demonstrate the effects of P1 infection on the Wnt signaling pathway. In vivo experiments, we demonstrated the down-regulatory effects of P1 infection in piglets and mice on the downstream components expression levels of Wnt signaling pathway, and the effects of Wnt signaling pathway activation on the pathogenesis of P1. In vitro studies, we found P1 infection down-regulated protein level of β-catenin and mRNA level of mmp2, prevented the β-catenin from entering into nucleus, abolished the TCF/LEF promoter activity, proved that P1 could inhibit the activation of Wnt signaling pathway in vitro. Finally, we found that VP1 of P1 virus also had the inhibitory effects on Wnt signaling pathway in vitro, elucidated the mechanism of P1’s inhibitory effects on the Wnt signaling pathway and offered the possibility that the suppression of Wnt signaling pathway was involved in the post-weaning multisystemic wasting syndrome (PMWS), laying a foundation for elucidating the pathogenesis of P1.

XAV939 Inhibits Intima Formation by Decreasing Vascular Smooth Muscle Cell Proliferation and Migration Through Blocking Wnt Signaling.

Excessive proliferation, migration, and oxidative stress of vascular smooth muscle cells (VSMCs) are key mechanisms involved in intima formation, which is the basic pathological process of in stent restenosis. This study aims at exploring the role of XAV939 in proliferation, migration, and reactive oxygen species (ROS) generation of VSMCs, and hence evaluating its effects on intima formation. Carotid artery ligation models for C57BL/6 mice were established and gave them different intervention: saline, XAV939, Axin2 overexpression adenovirus, and negative control adenovirus. The intima formation was assayed by intima area and intima/media ratio. To investigate the underlying mechanisms, primary rat VSMCs were cultured and treated with XAV939 and platelet-derived growth factor-BB. EdU, direct cell counting, cell wound-healing assay, and flow cytometry were used to measure proliferation, migration, cell cycle, apoptosis, and ROS generation of VSMCs, respectively. By Western blot, we examined proliferating cell nuclear antigen, Cyclin D1, Cyclin E, p21, β-actin, JNK, phosphorylated JNK, Axin2 and β-catenin expression. Immunofluorescence staining and confocal microscopy were conducted to detect translocation of β-catenin. XAV939 inhibited intima formation, which was exhibited by the loss of intima area and I/M ratio and attenuated proliferation, migration, and ROS generation, as well as promoted cell cycle arrest of VSMCs. Specifically, XAV939 inhibited Wnt pathway. XAV939 attenuates intima formation because of its inhibition of proliferation, migration, and apoptosis of VSMCs through suppression of Wnt signaling pathway.

PPARγ suppressed Wnt/β-catenin signaling pathway and its downstream effector SOX9 expression in gastric cancer cells.

Wnt signaling pathway activation plays a critical role in biological processes of tumor progression. SOX9 belongs to the sry-related high-mobility group box (SOX) family and is a key transcription factor in the development and differentiation of multiple cell lineages. The purpose of this study was to investigate whether suppression of Wnt signaling pathway by PPARγ gene affects target SOX9 gene expression. The pEGFP-N1-PPARγ overexpression recombinant plasmid was structured by molecular biology technology. The overexpression plasmid and empty vector pEGFP-N1 were transfected into three types of human gastric cancer cell lines, with different levels of differentiation, MKN-28, SGC-7901 and BGC-823. The PPARγ, β-catenin and SOX9 mRNA levels and proteins were examined by real-time PCR and Western blot analysis. The pEGFP-N1-PPARγ recombinant plasmid was constructed and transfected into MKN-28, SGC-7901 and BGC-823 successfully. High expression of PPARγ (p < 0.05) for transfection recombinant plasmid group induced obviously decreased expression of β-catenin (p < 0.05), whereas SOX9 expression decreased significantly (p < 0.05) compared with the transfection empty vector group and normal comparison group. PPARγ can suppress β-catenin expression in Wnt signaling pathway and its downstream effector SOX9 expression in gastric cancer cells.