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Calreticulin promotes EMT in pancreatic cancer via mediating Ca2+ dependent acute and chronic endoplasmic reticulum stress
Ming Dong
The first hospital of China Medical University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2685-1420
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at Journal of Experimental & Clinical Cancer Research.
Background: Our previous study showed that calreticulin (CRT) promoted EGF-induced epithelial-mesenchymal transition (EMT) in pancreatic cancer (PC) via Integrin/EGFR-ERK/MAPK signaling. We next investigated the novel signal pathway and molecular mechanism involving the oncogenic role of CRT in PC.
Methods: We investigated the potential role and mechanism of CRT in regulating intracellular free Ca2+ dependent acute and chronic endoplasmic reticulum stress (ERS)-induced EMT in PC in vitro and vivo.
Results: Thapsigargin (TG) induced acute ERS via increasing intracellular free Ca2+ in PC cells, which was reversed by CRT silencing. Additionally, CRT silencing inhibited TG-induced EMT in vitro by reversing TG-induced changes of the key proteins in EMT signaling (ZO-1, E-cadherin and Slug) and ERK/MAPK signaling (pERK). TG-promoted cell invasion and migration was also rescued by CRT silencing but enhanced by IRE1α silencing (one of the key stressors in unfolded protein response). Meanwhile, CRT was co-immunoprecipitated and co-localized with IRE1α in vitro and its silencing led to the chronic ERS via upregulating IRE1α independent of IRE1-XBP1 axis. Moreover, CRT silencing inhibited IRE1α silencing-promoted EMT, including inhibiting the activation of EMT and ERK/MAPK signaling and the promotion of cell mobility. In vivo, CRT silencing decreased subcutaneous tumor size and distant liver metastasis following with the increase of IRE1α expression. A negative relationship between CRT and IRE1α was also observed in clinical PC samples, which coordinately promoted the advanced clinical stages and poor prognosis of PC patients.
Conclusions: CRT promotes EMT in PC via mediating intracellular free Ca2+ dependent TG-induced acute ERS and IRE1α-mediated chronic ERS via Slug and ERK/MAPK signaling.
Keywords
Calreticulin, Intracellular free Ca2+, Endoplasmic reticulum stress, IRE1α, Epithelial mesenchymal transition, Pancreatic cancer
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This is a list of supplementary files associated with this preprint. Click to download.
- SupplementalMaterialFigure1.tif
Supplemental Figure 1. The statistic data of WB in Figure 1. A The quantified data of WB in Figure 2A. B The quantified data of WB in Figure 2B. C The quantified data of WB in Figure 2C. D The quantified data of WB in Figure 2D.
- SupplementalMaterialFigure2.tif
Supplemental Figure 2. IRE1α silencing enhanced TG-induced the increase of cell migration and invasion in vitro. A, B Cell invasion in Control, IRE1αsiRNA, TG and IRE1αsiRNA combing TG groups of Capan-2 (A) and SW1990 cells (B). C, D Cell migration in Control, IRE1αsiRNA, TG and IRE1αsiRNA combing TG groups of Capan-2 (C) and SW1990 cells (D). Data are shown as mean ± SD. *P < 0.05, **P < 0.01 versus control.
- SupplementalMaterialFigure3.tif
Supplemental Figure 3. The statistic data of WB in Figure 5. A The quantified data of WB in Figure 5A. B The quantified data of WB in Figure 5B. C The quantified data of WB in Figure 5C. D The quantified data of WB in Figure 5D.
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CURRENT STATUS: Published
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Posted 04 Sep, 2020
Published
On 07 Oct, 2020
No community comments so far
Editorial decision: Accept
On 06 Sep, 2020
Reviewer #1 agreed
On 04 Sep, 2020
Review #1 received
Received 04 Sep, 2020
Editor assigned
On 31 Aug, 2020
Reviewers invited
Invitations sent on 31 Aug, 2020
Submission checks complete
On 30 Aug, 2020
Editor invited
On 30 Aug, 2020
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Editorial decision: Minor revision
On 05 Jul, 2020
Review #1 received
Received 04 Jul, 2020
Review #2 received
Received 04 Jul, 2020
Reviewer #2 agreed
On 28 Jun, 2020
Reviewer #1 agreed
On 23 Jun, 2020
Editor assigned
On 22 Jun, 2020
Reviewers invited
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First submitted
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This preprint is under consideration at Journal of Experimental & Clinical Cancer Research. A preprint is 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
Calreticulin promotes EMT in pancreatic cancer via mediating Ca2+ dependent acute and chronic endoplasmic reticulum stress
Ming Dong
The first hospital of China Medical University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2685-1420
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
Version 2
Posted 04 Sep, 2020
Published
On 07 Oct, 2020
No community comments so far
Editorial decision: Accept
On 06 Sep, 2020
Reviewer #1 agreed
On 04 Sep, 2020
Review #1 received
Received 04 Sep, 2020
Editor assigned
On 31 Aug, 2020
Reviewers invited
Invitations sent on 31 Aug, 2020
Submission checks complete
On 30 Aug, 2020
Editor invited
On 30 Aug, 2020
No comments provided
Editorial decision: Minor revision
On 05 Jul, 2020
Review #1 received
Received 04 Jul, 2020
Review #2 received
Received 04 Jul, 2020
Reviewer #2 agreed
On 28 Jun, 2020
Reviewer #1 agreed
On 23 Jun, 2020
Editor assigned
On 22 Jun, 2020
Reviewers invited
Invitations sent on 22 Jun, 2020
First submitted
On 21 Jun, 2020
Submission checks complete
On 21 Jun, 2020
Editor invited
On 21 Jun, 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 version at Journal of Experimental & Clinical Cancer Research.
Background: Our previous study showed that calreticulin (CRT) promoted EGF-induced epithelial-mesenchymal transition (EMT) in pancreatic cancer (PC) via Integrin/EGFR-ERK/MAPK signaling. We next investigated the novel signal pathway and molecular mechanism involving the oncogenic role of CRT in PC.
Methods: We investigated the potential role and mechanism of CRT in regulating intracellular free Ca2+ dependent acute and chronic endoplasmic reticulum stress (ERS)-induced EMT in PC in vitro and vivo.
Results: Thapsigargin (TG) induced acute ERS via increasing intracellular free Ca2+ in PC cells, which was reversed by CRT silencing. Additionally, CRT silencing inhibited TG-induced EMT in vitro by reversing TG-induced changes of the key proteins in EMT signaling (ZO-1, E-cadherin and Slug) and ERK/MAPK signaling (pERK). TG-promoted cell invasion and migration was also rescued by CRT silencing but enhanced by IRE1α silencing (one of the key stressors in unfolded protein response). Meanwhile, CRT was co-immunoprecipitated and co-localized with IRE1α in vitro and its silencing led to the chronic ERS via upregulating IRE1α independent of IRE1-XBP1 axis. Moreover, CRT silencing inhibited IRE1α silencing-promoted EMT, including inhibiting the activation of EMT and ERK/MAPK signaling and the promotion of cell mobility. In vivo, CRT silencing decreased subcutaneous tumor size and distant liver metastasis following with the increase of IRE1α expression. A negative relationship between CRT and IRE1α was also observed in clinical PC samples, which coordinately promoted the advanced clinical stages and poor prognosis of PC patients.
Conclusions: CRT promotes EMT in PC via mediating intracellular free Ca2+ dependent TG-induced acute ERS and IRE1α-mediated chronic ERS via Slug and ERK/MAPK signaling.
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