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.
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Objective: Our study aims to investigate the mechanism of the miR-129-5p/SPN axis in clear cell renal cell carcinoma (ccRCC), providing a novel direction for the targeted therapy of ccRCC.
Methods: Bioinformatics methods were implemented to find the differentially expressed genes (DEGs) associated with ccRCC from TCGA database. qRT-PCR was performed to detect miR-129-5p and SPN mRNA expression, while western bot was carried out for the detection of protein expression of SPN. Bioinformatics analysis was used to predict the binding sites of miR-129-5p on SPN 3’UTR, while dual-luciferase assay was conducted to verify their binding relationship. CCK-8 assay, colony formation assay, wound healing assay and Transwell assay were employed to measure ccRCC cell proliferative ability, cell formation ability, cell migratory and invasive abilities. Flow cytometry was implemented to assess cell cycle and apoptosis.
Results: miR-129-5p exhibited a significantly down-regulated expression level in ccRCC, while SPN showed a remarkably up-regulated expression level. Overexpressed miR-129-5p inhibited ccRCC cell proliferative, invasive and migratory capacities while induced cell cycle arrest in G0/G1 phase and promoted cell apoptosis. Dual-luciferase assay confirmed that there was a binding relationship between miR-129-5p and SPN. Moreover, overexpressed miR-129-5p remarkably reduced SPN expression in cancer cells, weakened the promoting effect of SPN on cell proliferation, migration, invasion and cell cycle progress, and led to enhanced cell apoptotic activity.
Conclusion: Our study proves the regulatory effect of the miR-129-5p/SPN axis in ccRCC, and provides a novel potential target for precise treatment of patients with ccRCC.
Keywords
SPN, miR-129-5p, clear cell renal cell carcinoma, proliferation, migration and invasion, apoptosis
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CURRENT STATUS: Under Review
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Posted 21 Jan, 2021
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Review #2 received
Received 31 Jan, 2021
Reviewer #2 agreed
On 25 Jan, 2021
Review #1 received
Received 25 Jan, 2021
Editor assigned
On 08 Jan, 2021
Reviewers invited
Invitations sent on 08 Jan, 2021
Reviewer #1 agreed
On 08 Jan, 2021
Submission checks complete
On 08 Jan, 2021
Editor invited
On 08 Jan, 2021
No comments provided
Editorial decision: Major revision
On 29 Nov, 2020
Review #2 received
Received 26 Nov, 2020
Reviewer #2 agreed
On 18 Nov, 2020
Reviewer #1 agreed
On 16 Oct, 2020
Review #1 received
Received 16 Oct, 2020
Reviewers invited
Invitations sent on 15 Oct, 2020
Editor assigned
On 13 Oct, 2020
Submission checks complete
On 12 Oct, 2020
Editor invited
On 12 Oct, 2020
No comments provided
Editorial decision: Major revision
On 04 Aug, 2020
Review #1 received
Received 03 Aug, 2020
Review #2 received
Received 02 Aug, 2020
Reviewer #3 agreed
On 01 Aug, 2020
Reviewer #2 agreed
On 28 Jul, 2020
Reviewers invited
Invitations sent on 27 Jul, 2020
Reviewer #1 agreed
On 27 Jul, 2020
Editor assigned
On 21 Jul, 2020
Editor invited
On 20 Jul, 2020
Submission checks complete
On 19 Jul, 2020
First submitted
On 17 Jul, 2020
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Subject Areas
Cancer Biology
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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
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 3
Posted 21 Jan, 2021
No community comments so far
Review #2 received
Received 31 Jan, 2021
Reviewer #2 agreed
On 25 Jan, 2021
Review #1 received
Received 25 Jan, 2021
Editor assigned
On 08 Jan, 2021
Reviewers invited
Invitations sent on 08 Jan, 2021
Reviewer #1 agreed
On 08 Jan, 2021
Submission checks complete
On 08 Jan, 2021
Editor invited
On 08 Jan, 2021
No comments provided
Editorial decision: Major revision
On 29 Nov, 2020
Review #2 received
Received 26 Nov, 2020
Reviewer #2 agreed
On 18 Nov, 2020
Reviewer #1 agreed
On 16 Oct, 2020
Review #1 received
Received 16 Oct, 2020
Reviewers invited
Invitations sent on 15 Oct, 2020
Editor assigned
On 13 Oct, 2020
Submission checks complete
On 12 Oct, 2020
Editor invited
On 12 Oct, 2020
No comments provided
Editorial decision: Major revision
On 04 Aug, 2020
Review #1 received
Received 03 Aug, 2020
Review #2 received
Received 02 Aug, 2020
Reviewer #3 agreed
On 01 Aug, 2020
Reviewer #2 agreed
On 28 Jul, 2020
Reviewers invited
Invitations sent on 27 Jul, 2020
Reviewer #1 agreed
On 27 Jul, 2020
Editor assigned
On 21 Jul, 2020
Editor invited
On 20 Jul, 2020
Submission checks complete
On 19 Jul, 2020
First submitted
On 17 Jul, 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
Objective: Our study aims to investigate the mechanism of the miR-129-5p/SPN axis in clear cell renal cell carcinoma (ccRCC), providing a novel direction for the targeted therapy of ccRCC.
Methods: Bioinformatics methods were implemented to find the differentially expressed genes (DEGs) associated with ccRCC from TCGA database. qRT-PCR was performed to detect miR-129-5p and SPN mRNA expression, while western bot was carried out for the detection of protein expression of SPN. Bioinformatics analysis was used to predict the binding sites of miR-129-5p on SPN 3’UTR, while dual-luciferase assay was conducted to verify their binding relationship. CCK-8 assay, colony formation assay, wound healing assay and Transwell assay were employed to measure ccRCC cell proliferative ability, cell formation ability, cell migratory and invasive abilities. Flow cytometry was implemented to assess cell cycle and apoptosis.
Results: miR-129-5p exhibited a significantly down-regulated expression level in ccRCC, while SPN showed a remarkably up-regulated expression level. Overexpressed miR-129-5p inhibited ccRCC cell proliferative, invasive and migratory capacities while induced cell cycle arrest in G0/G1 phase and promoted cell apoptosis. Dual-luciferase assay confirmed that there was a binding relationship between miR-129-5p and SPN. Moreover, overexpressed miR-129-5p remarkably reduced SPN expression in cancer cells, weakened the promoting effect of SPN on cell proliferation, migration, invasion and cell cycle progress, and led to enhanced cell apoptotic activity.
Conclusion: Our study proves the regulatory effect of the miR-129-5p/SPN axis in ccRCC, and provides a novel potential target for precise treatment of patients with ccRCC.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
This is a list of supplementary files associated with this preprint. Click to download.
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