KEAP1-NRF2 , TWIST1, SNAI2, ZEB1, EMT, Melanooma

(Vuodelta 2017) Huom.Kirjoittajina ovat suomalaiset tiedemiehet!
https://www.ncbi.nlm.nih.gov/pubmed/27170270

Histol Histopathol. 2017 Feb;32(2):129-136. doi: 10.14670/HH-11-778. Epub 2016 May 12.An immunohistochemical study of NFE2L2, KEAP1 and 8-hydroxy-2'-deoxyguanosine and the EMT markers SNAI2, ZEB1 and TWIST1 in metastatic melanoma.Hintsala HR^1,2,3, Haapasaari KM⁴, Soini Y¹, Karihtala P³.

Tiivistelmä . 

Tausta: Tiedetään vain vähän redoxtasapainon säätelijöitten merkityksestä metastaattisessa melanoomassa, mutta jonkin verran on  kuitenkin  näyttöä  siitä, että on olemassa eräs linkki  solun redox-tilan ja epiteliaalisesta mesenkymaaliseksi transition (EMT) kesken.

• Abstract
• Role of redox balance regulators in metastatic melanomas, but there is some evidence for a link between epithelial-to-mesenchymal transition (EMT) and cellular redox status.

Menetelmät: Tutkijat vertailivat  NFE2L2-, KEAP1-, 8-OHdG-, TWIST1-, SNAI2- ja ZEB1- proteiinien  immunohistokemiallista ilmentymää primäärimelanomissa ja metastaaseissa  kohortista, jossa oli 23  naevus-luonta, 66 malignia melanoma ja 22 metastaasia. 

• METHODS:We compared the immunohistochemical expression of nuclear factor erythroid-2-related factor 2 (NFE2L2), Kelch-like ECH-associated protein 1 (KEAP1), 8-hydroxy-2'-deoxyguanosine (8-OHdG), TWIST1, SNAI2 and ZEB1 between primary melanomas and metastases in a cohort of 23 nevi, 66 malignant melanomas and 22 metastases.

Tulokset:  Tuman NFE2LL2-ilmentymä oli korkeampi  ja sytoplasminen KEAP1 matalampi metastaattisissa leesioissa kuin primäärimelanomakohdassa.  Tuman NFE2L2-ilmentymä assosioitui etäisten  metastaasien olemassaoloon  ja tuman TWIST1-ilmentymään.   Niiden potilaiden prognoosi oli äärimmäisen huono, joilla oli sekä NFE2L2- ja TWIST1-ilmentymät tumissa.
RESULTS:

• Nuclear NFE2L2 expression was higher (p=0.003) and cytoplasmic KEAP1 lower (p=0.026) in metastatic lesions than at primary sites. Nuclear NFE2L2 expression was associated with the presence of distant metastases (p=0.040) and with nuclear TWIST1 expression (p=0.002). Patients having both NFE2L2 and TWIST1 expression in nuclei had an extremely poor prognosis (p=0.0003). 

Monimuuttuja-analyysissä tuman TWIST1-ilmentymä  oli  itsenäinen ennuste huonommasta  prognoosista ja invasiivinen TWIST1/ZEB1-fenotyyppi osoitti huonompaa melanoomaspesifistä elossapysymistä.  Tuman 8-OHdG-ilemntymä oli matalampi  metastaasikohdissa kuin primäärileesioissa.

• In multivariate analysis nuclear TWIST1 expression was an independent predictor of a poorer prognosis (HR 2.99, 95% CI 1.17-7.69; p=0.023) and the invasive TWIST1/ZEB1 phenotype showed poorer melanoma-specific survival (HR 7.28, 95% CI 2.23-23.77; p=0.001). Nuclear expression of 8-OHdG (p=0.001) was lower at metastatic sites than in primary lesions.

 Yhteenveto:  EMT-signalointi ja KEAP1/NFL2L2-akseli ovat todennäköisesti sekaantuneet   malignin melanooman metastaattiseen  leviämiseen ja ilmeisesti niillä näyttää olevan  mahdollista vuorovaikutusta.

• CONCLUSIONS:EMT signalling and the KEAP1/NFE2L2-axis are likely to be involved in metastatic spread of malignant melanoma and also appear to have potential interactions.

PMID:

27170270

DOI:

10.14670/HH-11-778

Avainsanoja:

NFE2L2, nuclear factor erythroid-2-related factor 2.

KEAP1, Kelch-like ECH-associated protein 1.

8-OHdG, 8-hydroxy-2'-deoxyguanosine.

TWIST1,twist family bHLH transcription factor 1

https://www.ncbi.nlm.nih.gov/gene/7291.

SNAI2, Zinc finger protein SLUG, https://www.uniprot.org/uniprot/O43623

ZEB1, Zinc finger E-box binding homeobox 1 

 https://www.uniprot.org/uniprot/P37275.

 

 

ZC2HC alaryhmä 12. Nanos-geenit ja proteiinit. ( CCHC)2 zinc finger proteiinit, onkofetaalit antigeenit

• NANOS3 https://www.ncbi.nlm.nih.gov/gene/342977, ZC2HC12C, NANOS1L, NOS3
• NANOS2 https://www.ncbi.nlm.nih.gov/gene/339345 ,ZC2HC12B, NOS2
• NANOS1 https://www.ncbi.nlm.nih.gov/gene/340719, ZC2HC12A, NOS1, SPGF12

Cell Mol Life Sci. 2018 Jun;75(11):1929-1946. doi: 10.1007/s00018-018-2766-3. Epub 2018 Feb 3.

Nanos genes and their role in development and beyond.

De Keuckelaere E^1,2,3,4, Hulpiau P^1,2, Saeys Y^1,5, Berx G^3,4, van Roy F^6,7.

Abstract

The hallmark of Nanos proteins is their typical (CCHC)₂ zinc finger motif (zf-nanos). Animals have one to four nanos genes. For example, the fruit fly and demosponge have only one nanos gene, zebrafish and humans have three, and Fugu rubripes has four.

Nanos genes are mainly known for their evolutionarily preserved role in germ cell survival and pluripotency. Nanos proteins have been reported to bind the C-terminal RNA-binding domain of Pumilio to form a post-transcriptional repressor complex. Several observations point to a link between the miRNA-mediated repression complex and the Nanos/Pumilio complex. Repression of the E2F3 oncogene product is, indeed, mediated by cooperation between the Nanos/Pumilio complex and miRNAs. Another important interaction partner of Nanos is the CCR4-NOT deadenylase complex. Besides the tissue-specific contribution of Nanos proteins to normal development, their ectopic expression has been observed in several cancer cell lines and various human cancers. An inverse correlation between the expression levels of human Nanos1 and Nanos3 and E-cadherin was observed in several cancer cell lines. Loss of E-cadherin, an important cell-cell adhesion protein, contributes to tumor invasion and metastasis. Overexpression of Nanos3 induces epithelial-mesenchymal transition in lung cancer cell lines partly by repressing E-cadherin. Other than some most interesting data from Nanos knockout mice, little is known about mammalian Nanos proteins, and further research is needed. In this review, we summarize the main roles of Nanos proteins and discuss the emerging concept of Nanos proteins as oncofetal antigens.

KEYWORDS:

Cancer; Cancer testis antigen; Germ cell specification; Multiprotein complexes; Nanos; Phylogeny; Pumilio; RNA regulation; RNA-binding protein; pRb deficiency

PMID:

29397397

DOI:

10.1007/s00018-018-2766-3

(2) https://www.ncbi.nlm.nih.gov/pubmed/9049312

EMBO J. 1997 Feb 17;16(4):834-43.

A CCHC metal-binding domain in Nanos is essential for translational regulation.

Curtis D¹, Treiber DK, Tao F, Zamore PD, Williamson JR, Lehmann R.Abstract

The Drosophila Nanos protein is a localized repressor of hunchback mRNA translation in the early embryo, and is required for the establishment of the anterior-posterior body axis. Analysis of nanos mutants reveals that a small, evolutionarily conserved, C-terminal region is essential for Nanos function in vivo, while no other single portion of the Nanos protein is absolutely required. Within the C-terminal region are two unusual Cys-Cys-His-Cys (CCHC) motifs that are potential zinc-binding sites. Using absorption spectroscopy and NMR we demonstrate that the CCHC motifs each bind one equivalent of zinc with high affinity. nanos mutations disrupting metal binding at either of these two sites in vitro abolish Nanos translational repression activity in vivo. We show that full-length and C-terminal Nanos proteins bind to RNA in vitro with high affinity, but with little sequence specificity. Mutations affecting the hunchback mRNA target sites for Nanos-dependent translational repression were found to disrupt translational repression in vivo, but had little effect on Nanos RNA binding in vitro. Thus, the Nanos zinc domain does not specifically recognize target hunchback RNA sequences, but might interact with RNA in the context of a larger ribonucleoprotein complex.

PMID:

9049312

PMCID:

PMC1169684

Väitöskirja syöpätutkimuksen alalta EMT

http://hdl.handle.net/2077/35940
Tri Gisela Nilsson  

breast cancer epithelial-mesenchymal transition c-erbB2 E-cadherin NFI-C2 FoxF1 LOX

Abstract:

Epithelial-mesenchymal transition (EMT) is a developmental process defined by loss of epithelial characteristics and acquisition of mesenchymal phenotype. EMT or similar processes are also implicated in carcinoma cell invasion and the progression of breast carcinoma to metastasis. In a cell model system for mammary carcinogenesis it has previously been shown that signaling from the oncogenic receptor tyrosine kinase c-erbB2 (HER2), frequently overexpressed in mammary cancers, induces EMT. In this system, c-erbB2-induced EMT was significantly delayed by high cell-density and cell-cell-dissociation occurred before downregulation of the epithelial adhesion molecule E-cadherin. Loss of E-cadherin expression is generally viewed as a fundamental event in EMT. This thesis shows that ectopic expression of E-cadherin concomitant with c-erbB2 signaling did not hinder the progression of EMT. E-cadherin expressed in mesenchymal cells had a weaker attachment to the cytoskeleton, implicating that rearrangement of the cytoskeleton is an important mechanism in EMT-associated cell-cell-dissociation. Expression of dominant negative E-cadherin weakened cell-cell adhesion but did not enable EMT at high cell-density. These finding indicate that loss of E-cadherin is a consequence rather than a cause of EMT and that density-dependent inhibition of EMT is not mediated by E-cadherin. The expression of the transcription factor nuclear factor I-C2 (NFI-C2) is lost during mammary tumor progression and NFI-C2 has been shown to counteract EMT by repressing the transcription factor Forkhead box F1 (FoxF1). FoxF1 induces EMT and invasiveness in breast cancer cells. In this thesis, Affymetrix microarray was used to find oppositely regulated targets of NFI-C2 and FoxF1. The extracellular matrix enzyme lysyl oxidase (LOX) was found to be negatively regulated by NFI-C2 and positively regulated by FoxF1 and responsible for the increased invasiveness caused by FoxF1 overexpression. A signaling pathway was identified where FoxF1-induced upregulation of LOX activated focal adhesion kinase, subsequently suppressing Smad2 activity. In parallel, overexpression of FoxF1 activated the p38 MAPK signaling pathway. These findings give new insights into the regulation of signaling pathways known to be important during breast tumor progression. Based on the findings that NFI-C2 is lost during breast tumor progression and suppresses EMT, the prognostic value of NFI-C2 in a mixed cohort of breast cancer patients was investigated. NFI-C2 was found to be a powerful prognostic marker associated with good prognosis in breast cancer.