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Primary research
Anupam Chatterjee
North-Eastern Hill University School of Life Sciences
Corresponding Author
ORCiD: https://orcid.org/0000-0003-0486-0449
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Raw areca-nut (RAN) consumption induces oral, esophageal and gastric cancers which are significantly associated with the overexpression of pituitary tumor transforming gene1/Securin and chromosomal instability (CIN). Since the molecular mechanism underlying Securin upregulation remains unclear, this study is intended to investigate the association of Securin upregulation with Rb-E2F1 circuit and epigenetic histone (H3) modifications pattern both globally and in the promoter region of Securin gene.
Methods
Six groups of mice were used and in the treated group each mouse consumed 1 mg of RAN-extract with lime per day ad libitum in the drinking water for 60 days after which the dose was increased by 1 mg after every 60 days. Histopathological evaluation of stomach tissues was done and Securin expression was analysed by immunoblotting as well as by immunohistochemistry. ChIP-qPCR assay was performed for evaluating the recruitment of different histone modifications in the two regions of Securin promoter.
Results
All mice developed gastric cancer with Securin overexpression after 300 days of feeding. Immunohistochemistry data revealed hyperphosphorylation of Rb and upregulation of E2F1 in the RAN-treated samples. Increased trimethylation of H3 Lysine4 and acetylation of H3 Lysine9 and 18 both globally and in the promoter region of Securin gene was observed by increasing the level of lysine-N-methyltransferase2A, lysine-acetyltransferase, EP-300 and PCAF after RAN-treatment. ChIP-qPCR data show an increased recruitment of H3K4me3, H3K9ac and H3K18ac in the promoter of Securin gene after RAN-exposure
Conclusions
In light of the present results, it seems that RAN-mediated pRb-inactivation induced Securin upregulation, a putative E2F1 target, by inducing misregulation in chromatin remodeling in its promoter region which led to transcriptional activation and subsequently develop chromosomal instability. Therefore, the present results have led to the hypothesis that RAN-induced changes in the epigenetic landscape, Securin overexpression and subsequent elevation of chromosomal instability are probably a by-product of the inactivation of the pRb pathway.
Keywords
PTTG1/Securin gene, Histone methylation, Histone acetylation, Rb phosphorylation
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On 08 Jun, 2021
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On 07 Jun, 2021
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On 21 May, 2021
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Posted 05 Jun, 2021
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On 05 Jun, 2021
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Posted 21 May, 2021
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Editorial decision: Revise Before Review
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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
Anupam Chatterjee
North-Eastern Hill University School of Life Sciences
Corresponding Author
ORCiD: https://orcid.org/0000-0003-0486-0449
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 (no preprint)
Posted 08 Jun, 2021
First submitted
On 08 Jun, 2021
Submission checks complete
On 07 Jun, 2021
Editor invited
On 07 Jun, 2021
Editor assigned
On 07 Jun, 2021
Editorial decision: Revise Before Review
On 21 May, 2021
This version was not preprinted
Version (no preprint)
Posted 05 Jun, 2021
First submitted
On 05 Jun, 2021
This version was not preprinted
Version (no preprint)
Posted 21 May, 2021
This version was not preprinted
Version 1
Posted 21 May, 2021
No community comments so far
Editorial decision: Revise Before Review
On 20 May, 2021
Editor assigned
On 18 May, 2021
First submitted
On 17 May, 2021
Submission checks complete
On 17 May, 2021
Editor invited
On 17 May, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Raw areca-nut (RAN) consumption induces oral, esophageal and gastric cancers which are significantly associated with the overexpression of pituitary tumor transforming gene1/Securin and chromosomal instability (CIN). Since the molecular mechanism underlying Securin upregulation remains unclear, this study is intended to investigate the association of Securin upregulation with Rb-E2F1 circuit and epigenetic histone (H3) modifications pattern both globally and in the promoter region of Securin gene.
Methods
Six groups of mice were used and in the treated group each mouse consumed 1 mg of RAN-extract with lime per day ad libitum in the drinking water for 60 days after which the dose was increased by 1 mg after every 60 days. Histopathological evaluation of stomach tissues was done and Securin expression was analysed by immunoblotting as well as by immunohistochemistry. ChIP-qPCR assay was performed for evaluating the recruitment of different histone modifications in the two regions of Securin promoter.
Results
All mice developed gastric cancer with Securin overexpression after 300 days of feeding. Immunohistochemistry data revealed hyperphosphorylation of Rb and upregulation of E2F1 in the RAN-treated samples. Increased trimethylation of H3 Lysine4 and acetylation of H3 Lysine9 and 18 both globally and in the promoter region of Securin gene was observed by increasing the level of lysine-N-methyltransferase2A, lysine-acetyltransferase, EP-300 and PCAF after RAN-treatment. ChIP-qPCR data show an increased recruitment of H3K4me3, H3K9ac and H3K18ac in the promoter of Securin gene after RAN-exposure
Conclusions
In light of the present results, it seems that RAN-mediated pRb-inactivation induced Securin upregulation, a putative E2F1 target, by inducing misregulation in chromatin remodeling in its promoter region which led to transcriptional activation and subsequently develop chromosomal instability. Therefore, the present results have led to the hypothesis that RAN-induced changes in the epigenetic landscape, Securin overexpression and subsequent elevation of chromosomal instability are probably a by-product of the inactivation of the pRb pathway.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
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