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Desmoglein 2 depletion leads to increased migration and upregulation of the chemoattractant secretoneurin in melanoma cells.

Peitsch WK, Doerflinger Y, Fischer-Colbrie R, Huck V, Bauer AT, Utikal J, Goerdt S, Schneider SW - PLoS ONE (2014)

Bottom Line: During development and progression of malignant melanoma, an important role has been attributed to alterations of cell-cell adhesions, in particular, to a "cadherin switch" from E- to N-cadherin.In gene expression profiles, desmoglein 2 depletion was associated with overexpression of migration-related genes.The mechanism of desmoglein 2 impaired cell migration is mediated by downregulation of secretogranin II.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University Medical Center Mannheim, Heidelberg University, Mannheim, Germany ; Helmholtz Group for Cell Biology, German Cancer Research Center (DKFZ), Heidelberg, Germany.

ABSTRACT
During development and progression of malignant melanoma, an important role has been attributed to alterations of cell-cell adhesions, in particular, to a "cadherin switch" from E- to N-cadherin. We have previously shown that a subtype of melanoma cells express the desmosomal cadherin desmoglein 2 as non-junction-bound cell surface protein in addition to classical cadherins. To study the role of desmoglein 2 in melanoma cells, melanoma lines containing high endogenous amounts of desmoglein 2 were depleted of the protein by RNA interference. Transwell migration and scratch wounding assays showed markedly increased migration upon desmoglein 2 suppression whereas proliferation and viability remained unaltered. In gene expression profiles, desmoglein 2 depletion was associated with overexpression of migration-related genes. Strongest overexpression was found for secretogranin II which has not been reported in melanoma cells before. The bioactive peptide derived from secretogranin II, secretoneurin, is known to exert chemoattractive functions and was demonstrated here to stimulate melanoma cell migration. In summary, we show that desmoglein 2 expression attenuates migration of melanoma cells. The mechanism of desmoglein 2 impaired cell migration is mediated by downregulation of secretogranin II. Loss of desmoglein 2 increases expression of secretogranin II, followed by an enhanced migratory activity of melanoma cells. Our data add a new pathway of regulating melanoma cell migration related to a desmoglein 2-secretogranin II axis.

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Impact of Dsg2 on invasion.(A, B) Transwell invasion assays of Dsg2-depleted (“Dsg2 Ko”) or non-targeting siRNA-treated (“nt Ko”) MeWo and C32. Considerably more Dsg2 Ko than nt Ko MeWo had invaded after 24, 48 and 72 h (A); however, differences did not achieve significance. (B) In the line C32, the number of invaded cells was significantly increased upon Dsg2 knockdown after 48, 72 and 96 h. Bars: SD; * p≤0.05, ** p≤0.01. (C) TEER assay, showing no alteration of invasive properties upon Dsg2 depletion. A marked decrease in TEER was seen in cocultures of Dsg2 Ko, nt Ko or untreated MeWo and C32 with a MDCK-C7 monolayer after 48 and 72 h. However, no significant differences were noted between Dsg2-depleted and control cells.
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pone-0089491-g004: Impact of Dsg2 on invasion.(A, B) Transwell invasion assays of Dsg2-depleted (“Dsg2 Ko”) or non-targeting siRNA-treated (“nt Ko”) MeWo and C32. Considerably more Dsg2 Ko than nt Ko MeWo had invaded after 24, 48 and 72 h (A); however, differences did not achieve significance. (B) In the line C32, the number of invaded cells was significantly increased upon Dsg2 knockdown after 48, 72 and 96 h. Bars: SD; * p≤0.05, ** p≤0.01. (C) TEER assay, showing no alteration of invasive properties upon Dsg2 depletion. A marked decrease in TEER was seen in cocultures of Dsg2 Ko, nt Ko or untreated MeWo and C32 with a MDCK-C7 monolayer after 48 and 72 h. However, no significant differences were noted between Dsg2-depleted and control cells.

Mentions: Transwell invasion assays showed appreciably increased invasion of melanoma cells upon Dsg2 depletion. Ratios of Dsg2-depleted vs. non-targeting siRNA-treated MeWo migrated through a Matrigel-coated Transwell filter were 5.6, 7.3 and 4.7 after 24, 48 and 72 h. However, measured differences did not reach statistical significance, due to large variations (Fig. 4A). Significantly more Dsg2-depleted C32 than C32 controls had invaded and transmigrated after 48, 72 and 96 h, with ratios of 5.2, 4.4 and 3.8 (p = 0.014, p = 0.004 and p = 0.032; Fig. 4B).


Desmoglein 2 depletion leads to increased migration and upregulation of the chemoattractant secretoneurin in melanoma cells.

Peitsch WK, Doerflinger Y, Fischer-Colbrie R, Huck V, Bauer AT, Utikal J, Goerdt S, Schneider SW - PLoS ONE (2014)

Impact of Dsg2 on invasion.(A, B) Transwell invasion assays of Dsg2-depleted (“Dsg2 Ko”) or non-targeting siRNA-treated (“nt Ko”) MeWo and C32. Considerably more Dsg2 Ko than nt Ko MeWo had invaded after 24, 48 and 72 h (A); however, differences did not achieve significance. (B) In the line C32, the number of invaded cells was significantly increased upon Dsg2 knockdown after 48, 72 and 96 h. Bars: SD; * p≤0.05, ** p≤0.01. (C) TEER assay, showing no alteration of invasive properties upon Dsg2 depletion. A marked decrease in TEER was seen in cocultures of Dsg2 Ko, nt Ko or untreated MeWo and C32 with a MDCK-C7 monolayer after 48 and 72 h. However, no significant differences were noted between Dsg2-depleted and control cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928442&req=5

pone-0089491-g004: Impact of Dsg2 on invasion.(A, B) Transwell invasion assays of Dsg2-depleted (“Dsg2 Ko”) or non-targeting siRNA-treated (“nt Ko”) MeWo and C32. Considerably more Dsg2 Ko than nt Ko MeWo had invaded after 24, 48 and 72 h (A); however, differences did not achieve significance. (B) In the line C32, the number of invaded cells was significantly increased upon Dsg2 knockdown after 48, 72 and 96 h. Bars: SD; * p≤0.05, ** p≤0.01. (C) TEER assay, showing no alteration of invasive properties upon Dsg2 depletion. A marked decrease in TEER was seen in cocultures of Dsg2 Ko, nt Ko or untreated MeWo and C32 with a MDCK-C7 monolayer after 48 and 72 h. However, no significant differences were noted between Dsg2-depleted and control cells.
Mentions: Transwell invasion assays showed appreciably increased invasion of melanoma cells upon Dsg2 depletion. Ratios of Dsg2-depleted vs. non-targeting siRNA-treated MeWo migrated through a Matrigel-coated Transwell filter were 5.6, 7.3 and 4.7 after 24, 48 and 72 h. However, measured differences did not reach statistical significance, due to large variations (Fig. 4A). Significantly more Dsg2-depleted C32 than C32 controls had invaded and transmigrated after 48, 72 and 96 h, with ratios of 5.2, 4.4 and 3.8 (p = 0.014, p = 0.004 and p = 0.032; Fig. 4B).

Bottom Line: During development and progression of malignant melanoma, an important role has been attributed to alterations of cell-cell adhesions, in particular, to a "cadherin switch" from E- to N-cadherin.In gene expression profiles, desmoglein 2 depletion was associated with overexpression of migration-related genes.The mechanism of desmoglein 2 impaired cell migration is mediated by downregulation of secretogranin II.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University Medical Center Mannheim, Heidelberg University, Mannheim, Germany ; Helmholtz Group for Cell Biology, German Cancer Research Center (DKFZ), Heidelberg, Germany.

ABSTRACT
During development and progression of malignant melanoma, an important role has been attributed to alterations of cell-cell adhesions, in particular, to a "cadherin switch" from E- to N-cadherin. We have previously shown that a subtype of melanoma cells express the desmosomal cadherin desmoglein 2 as non-junction-bound cell surface protein in addition to classical cadherins. To study the role of desmoglein 2 in melanoma cells, melanoma lines containing high endogenous amounts of desmoglein 2 were depleted of the protein by RNA interference. Transwell migration and scratch wounding assays showed markedly increased migration upon desmoglein 2 suppression whereas proliferation and viability remained unaltered. In gene expression profiles, desmoglein 2 depletion was associated with overexpression of migration-related genes. Strongest overexpression was found for secretogranin II which has not been reported in melanoma cells before. The bioactive peptide derived from secretogranin II, secretoneurin, is known to exert chemoattractive functions and was demonstrated here to stimulate melanoma cell migration. In summary, we show that desmoglein 2 expression attenuates migration of melanoma cells. The mechanism of desmoglein 2 impaired cell migration is mediated by downregulation of secretogranin II. Loss of desmoglein 2 increases expression of secretogranin II, followed by an enhanced migratory activity of melanoma cells. Our data add a new pathway of regulating melanoma cell migration related to a desmoglein 2-secretogranin II axis.

Show MeSH
Related in: MedlinePlus