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Research article
Hong Zhang
Shengjing Hospital of China Medical University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-3327-1328
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
To investigate the effects of miR-200c, which targets fucosyltransferase 4 (FUT4), on the proliferation, migration and invasion of colon cancer cells and to further explore its mechanism.
Methods
We assessed the miR-200c and FUT4 mRNA levels in LoVo and SW480 cells by quantitative real-time polymerase chain reaction (qRT-PCR), and their correlation was analysed by Pearson correlation analysis. LipofectamineTM 2000 transfection reagent was used to transfect miR-200c mimic, FUT4 siRNA, FUT4 mimic and FUT4 mimic negative control into LoVo and SW480 cells, and RT-PCR was used to analyse the transfection efficiency. Cell Counting Kit-8 (CCK-8), colony formation and transwell assays were used to detect the migration, invasion and proliferation of LoVo and SW480 cells. Immunofluorescence was used to analyse the expression of the Ki-67 protein. Moreover, the expression of Wnt/β-catenin signalling pathway-related proteins was detected by western blots. A double luciferase experiment was performed to verify the targeting relationship between miR-200c and FUT4. In vivo, tumour growth and Wnt/β-catenin signalling pathway-related proteins were also analysed.
Results
Pearson correlation analysis showed that miR-200c and FUT4 were negatively correlated in LoVo and SW480 cells (correlation coefficients were -0.9046 and -0.9236, respectively). MiR-200c overexpression inhibited the proliferation, migration and invasion of LoVo cells by downregulating FUT4. The Ki67-positive cells and Wnt/β-catenin signalling pathway-related proteins were reduced in the miR-200c overexpression and FUT4 silencing groups. A bioinformatics analysis and a dual luciferase reporting system identified FUT4 as the target of miR-200c.
Conclusions
In conclusion, miR-200c overexpression inhibits FUT4 expression and downregulates the Wnt/β-catenin signalling pathway, thereby inhibiting the migration, invasion and proliferation of colon cancer cells.
Keywords
miR-200c, FUT4, colon cancer, Wnt/β-catenin pathway
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CURRENT STATUS: Published
View the published article at BMC Cancer.
No community comments so far
Submission checks complete
On 21 Nov, 2020
Editorial decision: Accept
On 18 Nov, 2020
No comments provided
Editor assigned
On 07 Nov, 2020
Submission checks complete
On 07 Nov, 2020
Editor invited
On 07 Nov, 2020
No comments provided
Editorial decision: Minor revision
On 28 Oct, 2020
Editor assigned
On 12 Oct, 2020
Submission checks complete
On 11 Oct, 2020
Editor invited
On 11 Oct, 2020
Version 2
Posted 11 Sep, 2020
No comments provided
Editorial decision: Minor revision
On 25 Sep, 2020
Review #2 received
Received 23 Sep, 2020
Reviewer #2 agreed
On 17 Sep, 2020
Reviewer #1 agreed
On 15 Sep, 2020
Review #1 received
Received 15 Sep, 2020
Reviewers invited
Invitations sent on 14 Sep, 2020
Editor assigned
On 08 Sep, 2020
Submission checks complete
On 07 Sep, 2020
Editor invited
On 07 Sep, 2020
No comments provided
Editorial decision: Major revision
On 20 Aug, 2020
Review #2 received
Received 17 Aug, 2020
Reviewer #2 agreed
On 29 Jul, 2020
Reviewer #3 agreed
On 29 Jul, 2020
Review #1 received
Received 24 Jun, 2020
Reviewer #1 agreed
On 04 Jun, 2020
Editor assigned
On 21 May, 2020
Reviewers invited
Invitations sent on 21 May, 2020
Submission checks complete
On 20 May, 2020
Editor invited
On 14 May, 2020
First submitted
On 06 May, 2020
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A new preprint design is coming!
We have improved readability, clarity, accessibility, and opportunities for engagement.
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
Hong Zhang
Shengjing Hospital of China Medical University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-3327-1328
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.
No community comments so far
Submission checks complete
On 21 Nov, 2020
Editorial decision: Accept
On 18 Nov, 2020
No comments provided
Editor assigned
On 07 Nov, 2020
Submission checks complete
On 07 Nov, 2020
Editor invited
On 07 Nov, 2020
No comments provided
Editorial decision: Minor revision
On 28 Oct, 2020
Editor assigned
On 12 Oct, 2020
Submission checks complete
On 11 Oct, 2020
Editor invited
On 11 Oct, 2020
Version 2
Posted 11 Sep, 2020
No comments provided
Editorial decision: Minor revision
On 25 Sep, 2020
Review #2 received
Received 23 Sep, 2020
Reviewer #2 agreed
On 17 Sep, 2020
Reviewer #1 agreed
On 15 Sep, 2020
Review #1 received
Received 15 Sep, 2020
Reviewers invited
Invitations sent on 14 Sep, 2020
Editor assigned
On 08 Sep, 2020
Submission checks complete
On 07 Sep, 2020
Editor invited
On 07 Sep, 2020
No comments provided
Editorial decision: Major revision
On 20 Aug, 2020
Review #2 received
Received 17 Aug, 2020
Reviewer #2 agreed
On 29 Jul, 2020
Reviewer #3 agreed
On 29 Jul, 2020
Review #1 received
Received 24 Jun, 2020
Reviewer #1 agreed
On 04 Jun, 2020
Editor assigned
On 21 May, 2020
Reviewers invited
Invitations sent on 21 May, 2020
Submission checks complete
On 20 May, 2020
Editor invited
On 14 May, 2020
First submitted
On 06 May, 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
To investigate the effects of miR-200c, which targets fucosyltransferase 4 (FUT4), on the proliferation, migration and invasion of colon cancer cells and to further explore its mechanism.
Methods
We assessed the miR-200c and FUT4 mRNA levels in LoVo and SW480 cells by quantitative real-time polymerase chain reaction (qRT-PCR), and their correlation was analysed by Pearson correlation analysis. LipofectamineTM 2000 transfection reagent was used to transfect miR-200c mimic, FUT4 siRNA, FUT4 mimic and FUT4 mimic negative control into LoVo and SW480 cells, and RT-PCR was used to analyse the transfection efficiency. Cell Counting Kit-8 (CCK-8), colony formation and transwell assays were used to detect the migration, invasion and proliferation of LoVo and SW480 cells. Immunofluorescence was used to analyse the expression of the Ki-67 protein. Moreover, the expression of Wnt/β-catenin signalling pathway-related proteins was detected by western blots. A double luciferase experiment was performed to verify the targeting relationship between miR-200c and FUT4. In vivo, tumour growth and Wnt/β-catenin signalling pathway-related proteins were also analysed.
Results
Pearson correlation analysis showed that miR-200c and FUT4 were negatively correlated in LoVo and SW480 cells (correlation coefficients were -0.9046 and -0.9236, respectively). MiR-200c overexpression inhibited the proliferation, migration and invasion of LoVo cells by downregulating FUT4. The Ki67-positive cells and Wnt/β-catenin signalling pathway-related proteins were reduced in the miR-200c overexpression and FUT4 silencing groups. A bioinformatics analysis and a dual luciferase reporting system identified FUT4 as the target of miR-200c.
Conclusions
In conclusion, miR-200c overexpression inhibits FUT4 expression and downregulates the Wnt/β-catenin signalling pathway, thereby inhibiting the migration, invasion and proliferation of colon cancer cells.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
This is a list of supplementary files associated with this preprint. Click to download.
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