See the published version of this preprint at Stem Cell Research & Therapy.
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
A new preprint design is coming!
We have improved readability, clarity, accessibility, and opportunities for engagement.
Research
Weilin Wang
Zhejiang University School of Medicine Second Affiliated Hospital
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Mesenchymal stem cells (MSCs) play an important role in tumour progression; concomitantly, MSCs also undergo profound changes in thetumour microenvironment (TME). These changes can directly impact the application and efficacy of MSC-based anti-tumour therapy. However, few studies have focused on the regulation of MSC fate in TME, which will limit the progress of MSC-based anti-tumour therapy. Herein, we investigated the effects of conditioned medium from human hepatocellular carcinoma cells (HCC-CM) on the phenotype and glucose metabolism of human adipose tissue-derived MSCs (hAT-MSCs).
Methods: hAT-MSCs were exposed to conditioned medium from Hep3B, Huh7 and HCCLM3cells for 4-8 weeks in vitro. Then, CCK-8 assay, EdU assay, Transwell assay, and flow cytometry were used to assess the alterations in cell phenotype in terms of cell proliferation, migration, invasion, cell cycle, and apoptosis. In addition, glucose metabolism was evaluated by related kits. Next,cells were subjected to withdrawal from HCC-CM for 2-4 weeks, andalterations in phenotype and glucose metabolism were reevaluatedin hAT-MSCs.Finally, the molecular mechanism was clarified by Western blotting.
Results: The results revealed that after exposure to HCC-CM, hAT-MSCs developed a stellate-shaped morphology. In association with cytoskeleton remodelling, hAT-MSCs showed enhanced capacities for migration and invasion, while cell proliferation was inhibited by regulating the cell cycle by downregulatingcyclins and cyclin-dependent kinases and activating the mitochondrial apoptosis pathway. In terms of glucose metabolism, our results showed mitochondrial dysfunction and elevated glycolysis of hAT-MSCs. However, interestingly, when hAT-MSCs were subjected towithdrawal from HCC-CM, the alterations in phenotype and glucose metabolism could be reversed. Further studies showed that these changes in hAT-MSCs may be regulated bythe ROS/MAPK/HIF-1α signalling pathway.
Conclusion:Taken together, the effects of long-term HCC-CM treatment on phenotype and glucose metabolism in hAT-MSCs are modest and largely reversible after withdrawal. This is the first report on the reversal of phenotype and glucose metabolism in tumour-associated MSCs (TA-MSCs), and it is anticipated that new insights into TA-MSCs will lead to the development of novel strategies for MSC-based anti-tumour therapy.
Keywords
Hepatocellular carcinoma, Conditioned medium, Human adipose tissue-derived mesenchymal stem cells, Cell phenotype, Glucose metabolism
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Loading...
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 Stem Cell Research & Therapy.
No community comments so far
Editorial decision: Accept
On 31 Oct, 2020
Review #1 received
Received 27 Oct, 2020
Reviewer #1 agreed
On 26 Oct, 2020
Reviewers invited
Invitations sent on 22 Oct, 2020
Editor assigned
On 21 Oct, 2020
Submission checks complete
On 20 Oct, 2020
Editor invited
On 20 Oct, 2020
No comments provided
Editorial decision: Minor Revision
On 10 Oct, 2020
Review #2 received
Received 03 Oct, 2020
Reviewer #2 agreed
On 29 Sep, 2020
Review #1 received
Received 22 Sep, 2020
Editor assigned
On 17 Sep, 2020
Reviewers invited
Invitations sent on 17 Sep, 2020
Reviewer #1 agreed
On 17 Sep, 2020
Submission checks complete
On 16 Sep, 2020
Editor invited
On 16 Sep, 2020
No comments provided
Editorial decision: Major Revision
On 23 Aug, 2020
Review #2 received
Received 22 Aug, 2020
Review #1 received
Received 14 Aug, 2020
Reviewer #2 agreed
On 10 Aug, 2020
Reviewer #1 agreed
On 08 Aug, 2020
Reviewers invited
Invitations sent on 06 Aug, 2020
Editor assigned
On 04 Aug, 2020
Submission checks complete
On 03 Aug, 2020
Editor invited
On 03 Aug, 2020
Version 1
Posted 14 May, 2020
No comments provided
Editorial decision: Major Revision
On 09 Jul, 2020
Review #3 received
Received 03 Jul, 2020
Review #1 received
Received 29 Jun, 2020
Review #2 received
Received 29 Jun, 2020
Reviewer #3 agreed
On 18 Jun, 2020
Reviewer #2 agreed
On 16 Jun, 2020
Reviewer #1 agreed
On 16 Jun, 2020
Reviewers invited
Invitations sent on 15 Jun, 2020
Editor assigned
On 14 May, 2020
Editor invited
On 13 May, 2020
Submission checks complete
On 08 May, 2020
First submitted
On 06 May, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Stem Cell & Developmental Cell Biology
Learn more about our company.
A new preprint design is coming!
We have improved readability, clarity, accessibility, and opportunities for engagement.
See the published version of this preprint at Stem Cell Research & Therapy.
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
Weilin Wang
Zhejiang University School of Medicine Second Affiliated Hospital
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 Stem Cell Research & Therapy.
No community comments so far
Editorial decision: Accept
On 31 Oct, 2020
Review #1 received
Received 27 Oct, 2020
Reviewer #1 agreed
On 26 Oct, 2020
Reviewers invited
Invitations sent on 22 Oct, 2020
Editor assigned
On 21 Oct, 2020
Submission checks complete
On 20 Oct, 2020
Editor invited
On 20 Oct, 2020
No comments provided
Editorial decision: Minor Revision
On 10 Oct, 2020
Review #2 received
Received 03 Oct, 2020
Reviewer #2 agreed
On 29 Sep, 2020
Review #1 received
Received 22 Sep, 2020
Editor assigned
On 17 Sep, 2020
Reviewers invited
Invitations sent on 17 Sep, 2020
Reviewer #1 agreed
On 17 Sep, 2020
Submission checks complete
On 16 Sep, 2020
Editor invited
On 16 Sep, 2020
No comments provided
Editorial decision: Major Revision
On 23 Aug, 2020
Review #2 received
Received 22 Aug, 2020
Review #1 received
Received 14 Aug, 2020
Reviewer #2 agreed
On 10 Aug, 2020
Reviewer #1 agreed
On 08 Aug, 2020
Reviewers invited
Invitations sent on 06 Aug, 2020
Editor assigned
On 04 Aug, 2020
Submission checks complete
On 03 Aug, 2020
Editor invited
On 03 Aug, 2020
Version 1
Posted 14 May, 2020
No comments provided
Editorial decision: Major Revision
On 09 Jul, 2020
Review #3 received
Received 03 Jul, 2020
Review #1 received
Received 29 Jun, 2020
Review #2 received
Received 29 Jun, 2020
Reviewer #3 agreed
On 18 Jun, 2020
Reviewer #2 agreed
On 16 Jun, 2020
Reviewer #1 agreed
On 16 Jun, 2020
Reviewers invited
Invitations sent on 15 Jun, 2020
Editor assigned
On 14 May, 2020
Editor invited
On 13 May, 2020
Submission checks complete
On 08 May, 2020
First submitted
On 06 May, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Stem Cell & Developmental Cell Biology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Stem Cell Research & Therapy.
Background: Mesenchymal stem cells (MSCs) play an important role in tumour progression; concomitantly, MSCs also undergo profound changes in thetumour microenvironment (TME). These changes can directly impact the application and efficacy of MSC-based anti-tumour therapy. However, few studies have focused on the regulation of MSC fate in TME, which will limit the progress of MSC-based anti-tumour therapy. Herein, we investigated the effects of conditioned medium from human hepatocellular carcinoma cells (HCC-CM) on the phenotype and glucose metabolism of human adipose tissue-derived MSCs (hAT-MSCs).
Methods: hAT-MSCs were exposed to conditioned medium from Hep3B, Huh7 and HCCLM3cells for 4-8 weeks in vitro. Then, CCK-8 assay, EdU assay, Transwell assay, and flow cytometry were used to assess the alterations in cell phenotype in terms of cell proliferation, migration, invasion, cell cycle, and apoptosis. In addition, glucose metabolism was evaluated by related kits. Next,cells were subjected to withdrawal from HCC-CM for 2-4 weeks, andalterations in phenotype and glucose metabolism were reevaluatedin hAT-MSCs.Finally, the molecular mechanism was clarified by Western blotting.
Results: The results revealed that after exposure to HCC-CM, hAT-MSCs developed a stellate-shaped morphology. In association with cytoskeleton remodelling, hAT-MSCs showed enhanced capacities for migration and invasion, while cell proliferation was inhibited by regulating the cell cycle by downregulatingcyclins and cyclin-dependent kinases and activating the mitochondrial apoptosis pathway. In terms of glucose metabolism, our results showed mitochondrial dysfunction and elevated glycolysis of hAT-MSCs. However, interestingly, when hAT-MSCs were subjected towithdrawal from HCC-CM, the alterations in phenotype and glucose metabolism could be reversed. Further studies showed that these changes in hAT-MSCs may be regulated bythe ROS/MAPK/HIF-1α signalling pathway.
Conclusion:Taken together, the effects of long-term HCC-CM treatment on phenotype and glucose metabolism in hAT-MSCs are modest and largely reversible after withdrawal. This is the first report on the reversal of phenotype and glucose metabolism in tumour-associated MSCs (TA-MSCs), and it is anticipated that new insights into TA-MSCs will lead to the development of novel strategies for MSC-based anti-tumour therapy.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Loading...