Wnt/β-catenin pathway regulates MGMT gene expression in cancer and inhibition of Wnt signalling prevents chemoresistance

Wickström, Malin; Dyberg, Cecilia; Milosevic, Jelena; Einvik, Christer; Calero, Raúl; Sveinbjørnsson, Baldur; Sandén, Emma; Darabi, Anna; Siesjö, Peter; Kool, Marcel; Kogner, Per; Baryawno, Ninib; Johnsen, John Inge

doi:10.1038/ncomms9904

Cited by 191 publications

(154 citation statements)

References 34 publications

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“…Protein extraction and Western blotting were performed as described earlier (29). Membranes (PVDF; Millipore) were incubated with antibodies against ROCK1 (1:1,000; 160 kDa), ROCK2 (1:1,000; 160 kDa), cyclin D1 (1:1,000; 36 kDa), TrkA (1:1,000; 140 kDa), MYCN (1:500; 62 kDa), β-actin (1:5,000; 45 kDa), GSK3β (1:1,000; 46 kDa), GSK3α/β phospho S21/S9 (1:1,000; 51 and 46 kDa), β3-tubulin (1:1,000; 55 kDa), MYPT1 (1:1,000; 140 kDa), phosphorylated MYPT1 (Thr696) (1:500; 140 kDa) (all from Cell Signaling), MYC phospho (T58/S62) (1:1,000; 57 to 65 kDa), GAPDH (1:10,000; 37 kDa), vinculin (1:10,000; 124 kDa) (all from Abcam), and MYCN (1:2,000; 62 kDa) (Santa Cruz Biotechnology).…”

Section: Discussionmentioning

confidence: 99%

Rho-associated kinase is a therapeutic target in neuroblastoma

Dyberg

Fransson

Andonova

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Neuroblastoma is a peripheral neural system tumor that originates from the neural crest and is the most common and deadly tumor of infancy. Here we show that neuroblastoma harbors frequent mutations of genes controlling the Rac/Rho signaling cascade important for proper migration and differentiation of neural crest cells during neuritogenesis. RhoA is activated in tumors from neuroblastoma patients, and elevated expression of Rho-associated kinase (ROCK)2 is associated with poor patient survival. Pharmacological or genetic inhibition of ROCK1 and 2, key molecules in Rho signaling, resulted in neuroblastoma cell differentiation and inhibition of neuroblastoma cell growth, migration, and invasion. Molecularly, ROCK inhibition induced glycogen synthase kinase 3β-dependent phosphorylation and degradation of MYCN protein. Small-molecule inhibition of ROCK suppressed MYCN-driven neuroblastoma growth in TH-MYCN homozygous transgenic mice and MYCN gene-amplified neuroblastoma xenograft growth in nude mice. Interference with Rho/Rac signaling might offer therapeutic perspectives for high-risk neuroblastoma.N euroblastoma is a childhood tumor of the peripheral sympathetic nervous system, causing 6% of all childhood cancers but 9% of deaths from malignant tumors in children (1, 2). Approximately half of patients present with high-risk disease characterized by unresectable primary lesions and multiple metastases (2, 3). Although survival in this group has improved, the majority of the tumors show resistance to therapy with poor patient survival despite intensive multimodal therapy, necessitating the search for new therapeutic options (2).Compared with adult tumors, pediatric cancers exhibit significantly fewer genomic aberrations and mutations. In neuroblastoma, somatically acquired amplification of MYCN, hemizygous deletions of 1p and 11q, and gain of 17q are the most common chromosomal aberrations associated with a poor prognosis (3). Exome-and whole-genome sequencing analysis of neuroblastoma has reported an overall low somatic mutation count (12 to 18, median 15), with ALK being the most frequently mutated gene (7 to 10%) (4-6). In addition, chromothripsis, PHOX2B, and ATRX mutations have also been detected in a subset of highrisk tumors (4,5).Neuritogenesis is initiated during embryogenesis by a transient population of cells called the neural crest. During embryonic development, neural crest cells migrate throughout the embryo and eventually differentiate into multiple cell types, such as neurons and glial cells of the peripheral nervous system, pigment cells, fibroblasts, smooth muscle cells, and odontoblasts. The failure of neural crest cells to differentiate can result in development of cancers such as neuroblastoma and melanoma (7). A combination of Wingless (Wnt), bone morphogenetic protein, and fibroblast growth factor (FGF) signals is required to induce the formation of the neural crest and to initiate migration of neural crest cells by acquiring cell motility through epithelialmesenchymal transition (8). The no...

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Section: Discussionmentioning

confidence: 99%

Rho-associated kinase is a therapeutic target in neuroblastoma

Dyberg

Fransson

Andonova

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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“…Further detailed studying of the molecular mechanisms involved in the dynamics of this pathway will pave the way for its clinical usage. Recently, Wnt pathway inhibitors have been studied successfully to block Wnt signaling in mouse models [63,64]. From this, it may be inferred that the bacterially expressed recombinant sFRP4 holds immense potential for in vivo cancer therapy.…”

Section: Discussionmentioning

confidence: 99%

Antagonizing canonical Wnt signaling pathway by recombinant human sFRP4 purified from E. coli and its implications in cancer therapy

Ghoshal

Ghosh

2016

Mol Cell Biochem

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The Wnt signaling pathway plays a predominant role in aberrant proliferation in myriad of cancers. In non-cancerous cells, Wnts are blocked by the secreted frizzled-related proteins (sFRPs) that are generally downregulated in cancer cells. We have purified and characterized bacterially expressed glutathione S-transferase-tagged SFRP4 from a novel clone generated from human cell origin. Cervical cancer (HeLa) and lung cancer (A549) cells, in which Wnt and associated genes were found to be expressed, were treated with the purified recombinant sFRP4, which revealed a significant dose-dependent cell growth inhibition up to 40 %. The current investigation on functionality of this bacterially produced recombinant sFRP4 in arresting cancer cell proliferation is the first of its kind, where G2/M phase arrest and early apoptosis were evident. Increase in phosphorylated β-catenin in sFRP4 treatment indicated inhibition of Wnt pathway, which was further confirmed by downregulation of pro-proliferative genes, namely cyclin D1, c-myc, and survivin. Functional activity of recombinant sFRP4 was further exploited in co-therapy module with chemotherapeutic drugs to decipher molecular events. Collectively, our study on purified recombinant sFRP4 from bacterial host holds great promise in targeting Wnt signaling for exploring new strategies to combat cancer.

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“…MGG8 or MGG6 patient-derived cells were cultured in NeuroCult NS-A proliferation kit (STEMCELL Technologies) with 10 μM of LGK974 (Selleckchem, S7143) diluted in DMSO (Wickstrom et al, 2015) or 50 mg/ml of the anti-VEGF B20 (Lu et al, 2012) for 72 hr. Then mRNA was extracted and RT-PCR performed as described above.…”

Section: Star⋆methodsmentioning

confidence: 99%

A Glial Signature and Wnt7 Signaling Regulate Glioma-Vascular Interactions and Tumor Microenvironment

et al. 2018

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SUMMARY Gliomas comprise heterogeneous malignant glial and stromal cells. While blood vessel co-option is a potential mechanism to escape anti-angiogenic therapy, the relevance of glial phenotype in this process is unclear. We show that Olig2+ oligodendrocyte precursor-like glioma cells invade by single-cell vessel co-option and preserve the blood-brain barrier (BBB). Conversely, Olig2-negative glioma cells form dense perivascular collections and promote angiogenesis and BBB breakdown, leading to innate immune cell activation. Experimentally, Olig2 promotes Wnt7b expression, a finding that correlates in human glioma profiling. Targeted Wnt7a/7b deletion or pharmacologic Wnt inhibition blocks Olig2+ glioma single-cell vessel co-option and enhances responses to temozolomide. Finally, Olig2 and Wnt7 become upregulated after anti-VEGF treatment in preclinical models and patients. Thus, glial-encoded pathways regulate distinct glioma-vascular microenvironmental interactions.

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Wnt/β-catenin pathway regulates MGMT gene expression in cancer and inhibition of Wnt signalling prevents chemoresistance

Cited by 191 publications

References 34 publications

Rho-associated kinase is a therapeutic target in neuroblastoma

Rho-associated kinase is a therapeutic target in neuroblastoma

Antagonizing canonical Wnt signaling pathway by recombinant human sFRP4 purified from E. coli and its implications in cancer therapy

A Glial Signature and Wnt7 Signaling Regulate Glioma-Vascular Interactions and Tumor Microenvironment

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