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Primary research
Lysophosphatidic Acid-induced EGFR Transactivation Promotes Gastric Cancer DNA Replication Through Up-Regulation of Geminin
Lifei Fan
Inner Mongolia University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2399-2447
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Lysophosphatidic acid (LPA) is one of the simplest active phospholipid molecules. Binding to its receptors on the cell surface, LPA initiates various intracellular signal cascades, involving in numerous biological processes, such as cell proliferation, migration, and apoptosis. If abnormalities occur in the processes of LPA production, receptor expression or signal transduction, it may induce certain diseases and even contribute to the occurrence, development and metastasis of cancer. However, whether the initiation of DNA replication is regulated by LPA has not yet been investigated.
Methods
First, diverse public databases were analyzed to explore the genetic abnormalities affecting geminin. Next, an LPA gradient treatment was performed on gastric cancer cells, followed by detecting geminin expression variation using western blot analysis. Finally, RNAi technology or inhibitors were used to block the biological activity of related factors in the GPCR induced EGFR transactivation signaling pathway to verify whether the effect of LPA evoked gastric cancer DNA replication is dependent on geminin upregulation.
Results
We found that LPA specifically up-regulated expression of an essential replication negative regulator geminin in early S phase in gastric cancer cell lines, and that the deletion of geminin selectively induced DNA re-replication. Neither of these phenomena has been observed in normal gastric epithelial cells, indicating LPA-induced geminin up-regulation is restricted to tumor cells. Using RNAi or specific inhibitors to block the activity of related factors in the signaling pathway, we found that LPA acts through LPAR3 and downstream coupled MMPs signaling to trans-activate EGFR, increasing the expression level of geminin in S phase. On the other hand, LPA stimulation induced the up-regulation of de-ubiquitinating enzyme 3 (DUB3) in a short time and inhibited the ubiquitination degradation of geminin to enhance geminin stability and positively regulate the DNA replication initiation in gastric cancer cells. Taken together, our results suggested that LPA mediated DNA replication and S-phase cell-cycle progression through a LPAR3/MMPs/EGFR/PI3K/mTOR signaling axis in gastric cancer.
Conclusions
Our research is for the first time to study the regulatory effect of LPA-induced EGFR transactivation in DNA replication of tumor cells, and to uncover a novel mechanism for regulating the stability of geminin through LPA and related downstream signaling pathways. All of which will provide potential targets for the development of signaling pathways and tumor cell-specific EGFR transactivation inhibitor for the treatment of gastric cancer.
Keywords
LPA, EGFR, transactivation, DNA replication, geminin
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On 01 Oct, 2020
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On 29 Sep, 2020
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On 29 Sep, 2020
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Received 29 Sep, 2020
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Received 29 Sep, 2020
Reviewer #6 agreed
On 28 Sep, 2020
Reviewer #7 agreed
On 28 Sep, 2020
Reviewer #8 agreed
On 28 Sep, 2020
Reviewer #9 agreed
On 28 Sep, 2020
Reviewer #10 agreed
On 28 Sep, 2020
Review #4 received
Received 23 Sep, 2020
Reviewer #2 agreed
On 23 Sep, 2020
Reviewer #3 agreed
On 23 Sep, 2020
Reviewer #4 agreed
On 23 Sep, 2020
Reviewer #5 agreed
On 23 Sep, 2020
Review #2 received
Received 23 Sep, 2020
Review #3 received
Received 23 Sep, 2020
Reviewers invited
Invitations sent on 22 Sep, 2020
Reviewer #1 agreed
On 22 Sep, 2020
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On 18 Sep, 2020
Editor invited
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First submitted
On 17 Sep, 2020
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This preprint is under consideration at Cancer Cell International. 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
Primary research
Lysophosphatidic Acid-induced EGFR Transactivation Promotes Gastric Cancer DNA Replication Through Up-Regulation of Geminin
Lifei Fan
Inner Mongolia University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2399-2447
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: Under Review
Version 1
Posted 22 Sep, 2020
No community comments so far
Editorial decision: Major revision
On 17 Oct, 2020
Review #11 received
Received 07 Oct, 2020
Review #5 received
Received 04 Oct, 2020
Review #6 received
Received 04 Oct, 2020
Review #9 received
Received 03 Oct, 2020
Reviewer #13 agreed
On 01 Oct, 2020
Review #8 received
Received 29 Sep, 2020
Review #7 received
Received 29 Sep, 2020
Reviewer #11 agreed
On 29 Sep, 2020
Reviewer #12 agreed
On 29 Sep, 2020
Review #10 received
Received 29 Sep, 2020
Review #1 received
Received 29 Sep, 2020
Reviewer #6 agreed
On 28 Sep, 2020
Reviewer #7 agreed
On 28 Sep, 2020
Reviewer #8 agreed
On 28 Sep, 2020
Reviewer #9 agreed
On 28 Sep, 2020
Reviewer #10 agreed
On 28 Sep, 2020
Review #4 received
Received 23 Sep, 2020
Reviewer #2 agreed
On 23 Sep, 2020
Reviewer #3 agreed
On 23 Sep, 2020
Reviewer #4 agreed
On 23 Sep, 2020
Reviewer #5 agreed
On 23 Sep, 2020
Review #2 received
Received 23 Sep, 2020
Review #3 received
Received 23 Sep, 2020
Reviewers invited
Invitations sent on 22 Sep, 2020
Reviewer #1 agreed
On 22 Sep, 2020
Editor assigned
On 18 Sep, 2020
Editor invited
On 17 Sep, 2020
First submitted
On 17 Sep, 2020
Submission checks complete
On 17 Sep, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Cancer Biology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Lysophosphatidic acid (LPA) is one of the simplest active phospholipid molecules. Binding to its receptors on the cell surface, LPA initiates various intracellular signal cascades, involving in numerous biological processes, such as cell proliferation, migration, and apoptosis. If abnormalities occur in the processes of LPA production, receptor expression or signal transduction, it may induce certain diseases and even contribute to the occurrence, development and metastasis of cancer. However, whether the initiation of DNA replication is regulated by LPA has not yet been investigated.
Methods
First, diverse public databases were analyzed to explore the genetic abnormalities affecting geminin. Next, an LPA gradient treatment was performed on gastric cancer cells, followed by detecting geminin expression variation using western blot analysis. Finally, RNAi technology or inhibitors were used to block the biological activity of related factors in the GPCR induced EGFR transactivation signaling pathway to verify whether the effect of LPA evoked gastric cancer DNA replication is dependent on geminin upregulation.
Results
We found that LPA specifically up-regulated expression of an essential replication negative regulator geminin in early S phase in gastric cancer cell lines, and that the deletion of geminin selectively induced DNA re-replication. Neither of these phenomena has been observed in normal gastric epithelial cells, indicating LPA-induced geminin up-regulation is restricted to tumor cells. Using RNAi or specific inhibitors to block the activity of related factors in the signaling pathway, we found that LPA acts through LPAR3 and downstream coupled MMPs signaling to trans-activate EGFR, increasing the expression level of geminin in S phase. On the other hand, LPA stimulation induced the up-regulation of de-ubiquitinating enzyme 3 (DUB3) in a short time and inhibited the ubiquitination degradation of geminin to enhance geminin stability and positively regulate the DNA replication initiation in gastric cancer cells. Taken together, our results suggested that LPA mediated DNA replication and S-phase cell-cycle progression through a LPAR3/MMPs/EGFR/PI3K/mTOR signaling axis in gastric cancer.
Conclusions
Our research is for the first time to study the regulatory effect of LPA-induced EGFR transactivation in DNA replication of tumor cells, and to uncover a novel mechanism for regulating the stability of geminin through LPA and related downstream signaling pathways. All of which will provide potential targets for the development of signaling pathways and tumor cell-specific EGFR transactivation inhibitor for the treatment of gastric cancer.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8