See the published version of this preprint at Cancer Cell International.
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.
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Primary research
Irina Krivosheeva
Mediko-geneticheskiy nauchniy centr
Corresponding Author
ORCiD: https://orcid.org/0000-0003-1743-3032
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
New approach for treating cancer is to influence on vital genes for tumors. A number of attempts have led to discovery candidate genes that are mean to be essential for surviving of tumor cells of different types. In this work, we tried to analyze genes that were previously found to be essential for melanoma surviving. We performed transient siRNA-mediated knockdown of UNC45A, STK11IP, RHPN2 and ZNFX1 in melanoma cell line A375. After that, we checked cell viability, proliferation and migrating rate in vitro.
Methods
For knockdown of the genes we used siRNA mediated interference. To deliver siRNA in cells we used transfection by Metafectene PRO. Cell viability and proliferation assessed by MTT assay. Cell migration was assessed by wound healing assay. Knockdown of genes was evaluated via RT-qPCR analysis.
Results
Our results indicate that knockdown of the genes leads to no statistically significant changes in cell survival according to viability assay. However, mobility assay shows that knockdown of each of the target genes accelerates the speed of cells migrating.
Conclusion
Our results do not confirm the hypothesis. Possible explaining of such controversial results could be in insufficient bioinformatical analysis or in need of multiplex knockdown of the genes.
Keywords
Melanoma, knockdown, viability, wound-healing, siRNA
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CURRENT STATUS: Published
View the published article at Cancer Cell International.
No community comments so far
Reviewer #1 agreed
On 19 Sep, 2020
Review #1 received
Received 19 Sep, 2020
Editor assigned
On 16 Sep, 2020
Reviewers invited
Invitations sent on 16 Sep, 2020
Editorial decision: Accept
On 16 Sep, 2020
Submission checks complete
On 15 Sep, 2020
Editor invited
On 15 Sep, 2020
Version 1
Posted 17 Jul, 2020
No comments provided
Editorial decision: Major revision
On 19 Aug, 2020
Review #2 received
Received 16 Aug, 2020
Reviewer #3 agreed
On 03 Aug, 2020
Reviewer #2 agreed
On 02 Aug, 2020
Reviewers invited
Invitations sent on 30 Jul, 2020
Reviewer #1 agreed
On 30 Jul, 2020
Review #1 received
Received 30 Jul, 2020
Editor assigned
On 20 Jul, 2020
Editor invited
On 19 Jul, 2020
Submission checks complete
On 15 Jul, 2020
First submitted
On 14 Jul, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Cancer Biology
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See the published version of this preprint at Cancer Cell International.
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
Primary research
Irina Krivosheeva
Mediko-geneticheskiy nauchniy centr
Corresponding Author
ORCiD: https://orcid.org/0000-0003-1743-3032
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 Cancer Cell International.
No community comments so far
Reviewer #1 agreed
On 19 Sep, 2020
Review #1 received
Received 19 Sep, 2020
Editor assigned
On 16 Sep, 2020
Reviewers invited
Invitations sent on 16 Sep, 2020
Editorial decision: Accept
On 16 Sep, 2020
Submission checks complete
On 15 Sep, 2020
Editor invited
On 15 Sep, 2020
Version 1
Posted 17 Jul, 2020
No comments provided
Editorial decision: Major revision
On 19 Aug, 2020
Review #2 received
Received 16 Aug, 2020
Reviewer #3 agreed
On 03 Aug, 2020
Reviewer #2 agreed
On 02 Aug, 2020
Reviewers invited
Invitations sent on 30 Jul, 2020
Reviewer #1 agreed
On 30 Jul, 2020
Review #1 received
Received 30 Jul, 2020
Editor assigned
On 20 Jul, 2020
Editor invited
On 19 Jul, 2020
Submission checks complete
On 15 Jul, 2020
First submitted
On 14 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
View the published article at Cancer Cell International.
Background
New approach for treating cancer is to influence on vital genes for tumors. A number of attempts have led to discovery candidate genes that are mean to be essential for surviving of tumor cells of different types. In this work, we tried to analyze genes that were previously found to be essential for melanoma surviving. We performed transient siRNA-mediated knockdown of UNC45A, STK11IP, RHPN2 and ZNFX1 in melanoma cell line A375. After that, we checked cell viability, proliferation and migrating rate in vitro.
Methods
For knockdown of the genes we used siRNA mediated interference. To deliver siRNA in cells we used transfection by Metafectene PRO. Cell viability and proliferation assessed by MTT assay. Cell migration was assessed by wound healing assay. Knockdown of genes was evaluated via RT-qPCR analysis.
Results
Our results indicate that knockdown of the genes leads to no statistically significant changes in cell survival according to viability assay. However, mobility assay shows that knockdown of each of the target genes accelerates the speed of cells migrating.
Conclusion
Our results do not confirm the hypothesis. Possible explaining of such controversial results could be in insufficient bioinformatical analysis or in need of multiplex knockdown of the genes.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
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