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Research article
Weiwen Long
Corresponding Author
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Background: p63, a member of the p53 gene family, is an important regulator for epithelial tissue growth and development. ∆Np63α is the main isoform of p63 and highly expressed in Non-melanoma skin cancer (NMSC). Extracellular signal-regulated kinase 3 (ERK3) is an atypical mitogen-activated protein kinase (MAPK) whose biochemical features and cellular regulation are distinct from those of conventional MAPKs such as ERK1/2. While ERK3 has been shown to be upregulated in lung cancers and head and neck cancers, in which it promotes cancer cell migration and invasion, little is known about the implication of ERK3 in NMSCs.
Methods: Fluorescent immunohistochemistry was performed to evaluate the expression levels of DNp63a and ERK3 in normal and NMSC specimens. Dunnett’s test was performed to compare mean fluorescence intensity (MFI, indicator of expression levels) of p63 or ERK3 between normal cutaneous samples and NMSC samples. A mixed effects (ANOVA) test was used to determine the correlation between DNp63a and ERK3 expression levels (MFI). The regulation of ERK3 by DNp63a was studied by qRT-PCR, Western blot and luciferase assay. The effect of ERK3 regulation by DNp63a on cell migration was measured by performing trans-well migration assay.
Results: The expression level of ∆Np63α is upregulated in NMSCs compared to normal tissue. ERK3 level is significantly upregulated in AK and SCC in comparison to normal tissue and there is a strong positive correlation between ∆Np63α and ERK3 expression in normal skin and skin specimens of patients with AK, SCC or BCC. Further, we found that ∆Np63α positively regulates ERK3 transcript and protein levels in A431 and HaCaT skin cells, underlying the upregulation of ERK3 expression and its positive correlation with ∆Np63α in NMSCs. Moreover, similar to the effect of ∆Np63α depletion, silencing ERK3 greatly enhanced A431 cell migration. Restoration of ERK3 expression under the condition of silencing ∆Np63α counteracted the increase in cell migration induced by the depletion of ∆Np63α. Mechanistically, ERK3 inhibits the phosphorylation of Rac1 G-protein and the formation of filopodia of A431 skin SCC cells.
Conclusions: ERK3 is positively regulated by ∆Np63α and mediates the role of ∆Np63α in suppressing cell migration in NMSC.
Keywords
ΔNp63α, ERK3, IHC, SCC, NMSC, migration
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View the published article at BMC Cancer.
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Received 09 Jul, 2020
Review #2 received
Received 09 Jul, 2020
Reviewer #2 agreed
On 01 Jul, 2020
Editor assigned
On 30 Jun, 2020
Reviewers invited
Invitations sent on 30 Jun, 2020
Reviewer #1 agreed
On 30 Jun, 2020
Submission checks complete
On 29 Jun, 2020
Editor invited
On 29 Jun, 2020
No comments provided
Editorial decision: Major revision
On 09 Apr, 2020
Review #2 received
Received 18 Mar, 2020
Review #1 received
Received 18 Mar, 2020
Reviewer #2 agreed
On 09 Mar, 2020
Reviewer #1 agreed
On 25 Feb, 2020
Editor assigned
On 12 Feb, 2020
Reviewers invited
Invitations sent on 12 Feb, 2020
Submission checks complete
On 12 Feb, 2020
Editor invited
On 12 Feb, 2020
First submitted
On 04 Feb, 2020
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See the published version of this preprint at BMC Cancer.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
Weiwen Long
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at BMC Cancer.
Version 2
Posted 02 Jul, 2020
No community comments so far
Review #1 received
Received 09 Jul, 2020
Review #2 received
Received 09 Jul, 2020
Reviewer #2 agreed
On 01 Jul, 2020
Editor assigned
On 30 Jun, 2020
Reviewers invited
Invitations sent on 30 Jun, 2020
Reviewer #1 agreed
On 30 Jun, 2020
Submission checks complete
On 29 Jun, 2020
Editor invited
On 29 Jun, 2020
No comments provided
Editorial decision: Major revision
On 09 Apr, 2020
Review #2 received
Received 18 Mar, 2020
Review #1 received
Received 18 Mar, 2020
Reviewer #2 agreed
On 09 Mar, 2020
Reviewer #1 agreed
On 25 Feb, 2020
Editor assigned
On 12 Feb, 2020
Reviewers invited
Invitations sent on 12 Feb, 2020
Submission checks complete
On 12 Feb, 2020
Editor invited
On 12 Feb, 2020
First submitted
On 04 Feb, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at BMC Cancer.
Background: p63, a member of the p53 gene family, is an important regulator for epithelial tissue growth and development. ∆Np63α is the main isoform of p63 and highly expressed in Non-melanoma skin cancer (NMSC). Extracellular signal-regulated kinase 3 (ERK3) is an atypical mitogen-activated protein kinase (MAPK) whose biochemical features and cellular regulation are distinct from those of conventional MAPKs such as ERK1/2. While ERK3 has been shown to be upregulated in lung cancers and head and neck cancers, in which it promotes cancer cell migration and invasion, little is known about the implication of ERK3 in NMSCs.
Methods: Fluorescent immunohistochemistry was performed to evaluate the expression levels of DNp63a and ERK3 in normal and NMSC specimens. Dunnett’s test was performed to compare mean fluorescence intensity (MFI, indicator of expression levels) of p63 or ERK3 between normal cutaneous samples and NMSC samples. A mixed effects (ANOVA) test was used to determine the correlation between DNp63a and ERK3 expression levels (MFI). The regulation of ERK3 by DNp63a was studied by qRT-PCR, Western blot and luciferase assay. The effect of ERK3 regulation by DNp63a on cell migration was measured by performing trans-well migration assay.
Results: The expression level of ∆Np63α is upregulated in NMSCs compared to normal tissue. ERK3 level is significantly upregulated in AK and SCC in comparison to normal tissue and there is a strong positive correlation between ∆Np63α and ERK3 expression in normal skin and skin specimens of patients with AK, SCC or BCC. Further, we found that ∆Np63α positively regulates ERK3 transcript and protein levels in A431 and HaCaT skin cells, underlying the upregulation of ERK3 expression and its positive correlation with ∆Np63α in NMSCs. Moreover, similar to the effect of ∆Np63α depletion, silencing ERK3 greatly enhanced A431 cell migration. Restoration of ERK3 expression under the condition of silencing ∆Np63α counteracted the increase in cell migration induced by the depletion of ∆Np63α. Mechanistically, ERK3 inhibits the phosphorylation of Rac1 G-protein and the formation of filopodia of A431 skin SCC cells.
Conclusions: ERK3 is positively regulated by ∆Np63α and mediates the role of ∆Np63α in suppressing cell migration in NMSC.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
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