| 1. |
- Jenei, Veronika, et al.
(författare)
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A t-butyloxycarbonyl-modified Wnt5a-derived hexapeptide functions as a potent antagonist of Wnt5a-dependent melanoma cell invasion.
- 2009
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Ingår i: Proceedings of the National Academy of Sciences. - : Proceedings of the National Academy of Sciences. - 1091-6490 .- 0027-8424. ; 106:46, s. 19473-19478
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Tidskriftsartikel (refereegranskat)abstract
- The influential role of Wnt5a in tumor progression underscores the requirement for developing molecules that can target Wnt5a-mediated cellular responses. In the aggressive skin cancer, melanoma, elevated Wnt5a expression promotes cell motility and drives metastasis. Two approaches can be used to counteract these effects: inhibition of Wnt5a expression or direct blockade of Wnt5a signaling. We have investigated both options in the melanoma cell lines, A2058 and HTB63. Both express Frizzled-5, which has been implicated as the receptor for Wnt5a in melanoma cells. However, only the HTB63 cell line expresses and secretes Wnt5a. In these cells, the cytokine, TGFbeta1, controlled the expression of Wnt5a, but due to the unpredictable effects of TGFbeta1 signaling on melanoma cell motility, targeting Wnt5a signaling via TGFbeta1 was an unsuitable strategy to pursue. We therefore attempted to target Wnt5a signaling directly. Exogenous Wnt5a stimulation of A2058 cells increased adhesion, migration and invasion, all crucial components of tumor metastasis, and the Wnt5a-derived N-butyloxycarbonyl hexapeptide (Met-Asp-Gly-Cys-Glu-Leu; 0.766 kDa) termed Box5, abolished these responses. Box5 also inhibited the basal migration and invasion of Wnt5a-expressing HTB63 melanoma cells. Box5 antagonized the effects of Wnt5a on melanoma cell migration and invasion by directly inhibiting Wnt5a-induced protein kinase C and Ca(2+) signaling, the latter of which we directly demonstrate to be essential for cell invasion. The Box5 peptide directly inhibits Wnt5a signaling, representing an approach to anti-metastatic therapy for otherwise rapidly progressive melanoma, and for other Wnt5a-stimulated invasive cancers.
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| 2. |
- Linnskog, Rickard, et al.
(författare)
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Demonstration of a WNT5A-IL-6 positive feedback loop in melanoma cells : Dual interference of this loop more effectively impairs melanoma cell invasion
- 2016
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Ingår i: Oncotarget. - : Impact Journals, LLC. - 1949-2553. ; 7:25, s. 37790-37802
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Tidskriftsartikel (refereegranskat)abstract
- Increased expression and signalling of WNT5A and interleukin-6 (IL-6) have both been shown to promote melanoma progression. Here, we investigated the proposed existence of a WNT5A-IL-6 positive feedback loop that drives melanoma migration and invasion. First, the HOPP algorithm revealed that the invasive phenotype of cultured melanoma cells was significantly correlated with increased expression of WNT5A or IL-6. In three invasive melanoma cell lines, endogenous WNT5A protein expression was related to IL-6 protein secretion. Knockdown with anti-IL-6 siRNAs or treating WM852 melanoma cells with a neutralising anti-IL-6 antibody reduced WNT5A protein expression. Conversely, the silencing of WNT5A expression by WNT5A siRNAs or treating WM852 melanoma cells with Box5 (a WNT5A antagonist) significantly reduced IL-6 secretion. Interestingly, these effects occurred at the protein level but not at the transcriptional levels. Functionally, we demonstrated that combined siRNA knockdown of WNT5A and IL-6 expression or the simultaneous inhibition of WNT5A and IL-6 signalling inhibited melanoma cell invasion more effectively than suppressing each factor individually. Together, our results demonstrate that WNT5A and IL-6 are connected through a positive feedback loop in melanoma cells and that the combined targeting of both molecules could serve as an effective therapeutic means to reduce melanoma metastasis.
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| 3. |
- Linnskog, Rickard
(författare)
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Inhibition of WNT5A Signaling and Function in Malignant Melanoma
- 2014
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- The life-threatening disease of malignant melanoma is one of the fastest growing forms of cancer, affecting an estimated number of 200,000 patients worldwide every year. Although highly curable if detected early, once the melanoma has metastasized there are few treatment options and a median survival rate of only 6-9 months. Consequently, there is an imperative need to identify the factors that drive melanoma metastasis and to develop means of interfering with their actions. Elevated WNT5A expression has been significantly correlated with increased metastatic potential of melanoma cells as well as reduced patient survival. In this thesis we investigate the possibility of impairing melanoma cell motility by targeting WNT5A signaling. Our results demonstrate that TGFβ is an unsuitable target for blocking WNT5A signaling since it mediates unpredictable effects on melanoma cell invasion. Instead, we show that the t-butyloxycarbonyl-modified WNT5A-derived hexapeptide Box5 functions a potent antagonist of WNT5A-induced signaling, migration and invasion in melanoma cells. Moreover, we demonstrate that IL-6 can drive melanoma cell motility through p38α-MAPK-dependent up-regulation of WNT5A expression and that targeting IL-6, in contrast to TGFβ, reduces melanoma cell invasion. We furthermore reveal the existence of a WNT5A-IL-6 positive feedback loop in melanoma cells that augments their invasive capacity. Concomitantly, we show that combined targeting of IL-6 and WNT5A within this loop more effectively impairs melanoma cell invasion. Taken together, our results demonstrate the possibilities of antagonizing WNT5A signaling directly (Box5) or indirectly (targeting of IL-6), which if combined can provide the basis for future therapeutic intervention of melanoma metastasis.
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| 4. |
- Linnskog, Rickard, et al.
(författare)
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Interleukin-6 drives melanoma cell motility through p38α-MAPK-dependent up-regulation of WNT5A expression.
- 2014
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Ingår i: Molecular Oncology. - : Wiley. - 1574-7891. ; 8:8, s. 1365-1378
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Tidskriftsartikel (refereegranskat)abstract
- Extensive research has demonstrated a tumor-promoting role of increased WNT5A expression in malignant melanoma. However, very little light has been shed upon how WNT5A expression is up-regulated in melanoma. A potential regulator of WNT5A expression is the pro-inflammatory cytokine Interleukin (IL)-6, which shares the ability of WNT5A to increase melanoma cell invasion. Here, we investigate whether IL-6 can promote melanoma cell motility through an increased expression of WNT5A. We clearly demonstrate that the WNT5A-antagonistic peptide Box5 could inhibit IL-6-induced melanoma cell migration and invasion. Furthermore, IL-6 stimulation of the human melanoma cell lines HTB63 and A375 increased the expression of WNT5A in a dose-dependent manner. To identify the signaling mechanism responsible for this up-regulation, we explored the involvement of the three main signals induced by IL-6; STAT3, Akt and ERK 1/2. Of these, only STAT3 was activated by IL-6 in the melanoma cell lines tested. However, the STAT3 inhibitor S3I-201 failed to inhibit IL-6-induced WNT5A up-regulation in HTB63 and A375 cells. Nor did STAT3 siRNA silencing affect the expression of WNT5A. In search of an alternative signaling mechanism, we detected IL-6-induced activation of p38-MAPK in HTB63 and A375 cells. The p38-MAPK inhibitor SB203580 abolished the IL-6-induced WNT5A up-regulation and blocked IL-6-induced melanoma cell invasion. The latter effect could be rescued by the addition of recombinant WNT5A. Notably, immunoprecipitation analysis revealed that only the p38α-MAPK isoform was activated by IL-6, and subsequent siRNA silencing of p38α-MAPK abolished the IL-6-induced up-regulation of WNT5A. Taken together, we demonstrate a novel link between the two melanoma pro-metastatic agents IL-6 and WNT5A explaining how IL-6 can increase melanoma cell invasion and thus promote the metastatic process. This finding provides a basis for future therapeutic intervention of melanoma progression.
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| 5. |
- Moradi, Farnaz, et al.
(författare)
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Dual mechanisms of action of the RNA-binding protein human antigen R explains its regulatory effect on melanoma cell migration.
- 2016
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Ingår i: Translational Research. - : Elsevier BV. - 1878-1810 .- 1931-5244. ; 172, s. 45-60
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Tidskriftsartikel (refereegranskat)abstract
- Overexpression of wingless-type MMTV integration site family 5A (WNT5A) plays a significant role in melanoma cancer progression; however, the mechanism(s) involved remains unknown. In breast cancer, the human antigen R (HuR) has been implicated in the regulation of WNT5A expression. Here, we demonstrate that endogenous expression of WNT5A correlates with levels of active HuR in HTB63 and WM852 melanoma cells and that HuR binds to WNT5A messenger RNA in both cell lines. Although the HuR inhibitor MS-444 significantly impaired migration in both melanoma cell lines, it reduced WNT5A expression only in HTB63 cells, as did small interfering RNA knockdown of HuR. Consistent with this finding, MS-444-induced inhibition of HTB63 cell migration was restored by the addition of recombinant WNT5A, whereas MS-444-induced inhibition of WM852 cell migration was restored by the addition of recombinant matrix metalloproteinase-9, another HuR-regulated protein. Clearly, HuR positively regulates melanoma cell migration via at least 2 distinct mechanisms making HuR an attractive therapeutic target for halting melanoma dissemination.
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