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Research article
Zhongliang Zheng
College of Life Sciences in Wuhan University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2861-5720
License:
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Background Based on its low toxicity, arginine starvation therapy has the potential to treat those malignant tumors that can’t be treated by surgery. Arginine deiminase (ADI; EC 3.5.3.6) gene is indicated to be an ideal cancer-suppressor gene. ADI expressed in cytosol displayed higher oncolytic efficiency than ADI-PEG20 (Pegylated Arginine Deiminase by PEG 20,000)[1]. However, it is still unknown whether cytosolic ADI has the same function mechanism as ADI-PEG20 or other underlying mechanisms in cells.
Methods The interaction of ADI and other protein factors was screened by yeast hybrid, and verified by co-immunoprecipitation and immunofluorescent staining. The effect of ADI inhibiting ferritin light-chain domain (FTL) on mitochondria damage was evaluated by site-directed mutation and flow cytometry. The apoptosis pathway of mitochondria control was analyzed by Western Blot and real-time PCR. The effect of p53 expression on cancer cell death was assessed by siTP53 transfection. The chromatin autophagy was explored by immunofluorescent staining and Western Blot.
Results ADI expressed in cytosol inhibited the activity of cytosolic ferritin through interacting with FTL. The inactive mutant of ADI still aroused the apoptosis of some cells through mitochondria damage. Arginine deprivation also induced the expression increase of p53 and p53AIP1, which aggravated cellular mitochondria damage. Chromatin autophagy appeared at the later stage of arginine deprivation. DNA damage came along with the whole process of arginine starvation. Histone 3 (H3) was found in autophagosomes, which implied that cancer cells try to utilize the arginine in histones to survive during arginine starvation.
Conclusions Mitochondria damage is the major mechanism for ADI expressed in cytosol to induce cancer cell death. Chromatophagy accumulation not only drives cancer cell to utilize histone arginine but also speeds up cancer cell death at the later time point of arginine deprivation.
Keywords
Arginine deprivation, arginine deiminase, apoptosis, mitochondria damage, chromotin autophagy
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Received 25 Feb, 2020
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Received 18 Feb, 2020
Review #1 received
Received 18 Feb, 2020
Reviewer #5 agreed
On 14 Feb, 2020
Reviewer #6 agreed
On 14 Feb, 2020
Reviewer #3 agreed
On 11 Feb, 2020
Reviewer #4 agreed
On 11 Feb, 2020
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Reviewer #1 agreed
On 11 Feb, 2020
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On 11 Feb, 2020
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A new preprint design is coming!
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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
Zhongliang Zheng
College of Life Sciences in Wuhan University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-2861-5720
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
Editorial decision: Accept
On 01 Jul, 2020
Editor assigned
On 29 Jun, 2020
Submission checks complete
On 28 Jun, 2020
Editor invited
On 28 Jun, 2020
No comments provided
Editorial decision: Minor revision
On 26 Jun, 2020
Editor assigned
On 23 Jun, 2020
Submission checks complete
On 22 Jun, 2020
Editor invited
On 22 Jun, 2020
No comments provided
Editorial decision: Minor revision
On 17 Jun, 2020
Editor assigned
On 15 Jun, 2020
Submission checks complete
On 14 Jun, 2020
Editor invited
On 14 Jun, 2020
No comments provided
Editorial decision: Minor revision
On 03 Jun, 2020
Editor assigned
On 02 Jun, 2020
Submission checks complete
On 01 Jun, 2020
Editor invited
On 01 Jun, 2020
Version 2
Posted 17 Apr, 2020
No comments provided
Editorial decision: Minor revision
On 09 May, 2020
Review #2 received
Received 09 May, 2020
Review #3 received
Received 18 Apr, 2020
Reviewer #3 agreed
On 17 Apr, 2020
Review #1 received
Received 15 Apr, 2020
Editor assigned
On 14 Apr, 2020
Reviewers invited
Invitations sent on 14 Apr, 2020
Reviewer #1 agreed
On 14 Apr, 2020
Reviewer #2 agreed
On 14 Apr, 2020
Submission checks complete
On 13 Apr, 2020
Editor invited
On 13 Apr, 2020
No comments provided
Editorial decision: Major revision
On 02 Mar, 2020
Review #4 received
Received 01 Mar, 2020
Review #3 received
Received 25 Feb, 2020
Review #2 received
Received 18 Feb, 2020
Review #1 received
Received 18 Feb, 2020
Reviewer #5 agreed
On 14 Feb, 2020
Reviewer #6 agreed
On 14 Feb, 2020
Reviewer #3 agreed
On 11 Feb, 2020
Reviewer #4 agreed
On 11 Feb, 2020
Reviewers invited
Invitations sent on 11 Feb, 2020
Reviewer #1 agreed
On 11 Feb, 2020
Reviewer #2 agreed
On 11 Feb, 2020
Editor assigned
On 04 Feb, 2020
Submission checks complete
On 03 Feb, 2020
Editor invited
On 03 Feb, 2020
First submitted
On 02 Feb, 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 Based on its low toxicity, arginine starvation therapy has the potential to treat those malignant tumors that can’t be treated by surgery. Arginine deiminase (ADI; EC 3.5.3.6) gene is indicated to be an ideal cancer-suppressor gene. ADI expressed in cytosol displayed higher oncolytic efficiency than ADI-PEG20 (Pegylated Arginine Deiminase by PEG 20,000)[1]. However, it is still unknown whether cytosolic ADI has the same function mechanism as ADI-PEG20 or other underlying mechanisms in cells.
Methods The interaction of ADI and other protein factors was screened by yeast hybrid, and verified by co-immunoprecipitation and immunofluorescent staining. The effect of ADI inhibiting ferritin light-chain domain (FTL) on mitochondria damage was evaluated by site-directed mutation and flow cytometry. The apoptosis pathway of mitochondria control was analyzed by Western Blot and real-time PCR. The effect of p53 expression on cancer cell death was assessed by siTP53 transfection. The chromatin autophagy was explored by immunofluorescent staining and Western Blot.
Results ADI expressed in cytosol inhibited the activity of cytosolic ferritin through interacting with FTL. The inactive mutant of ADI still aroused the apoptosis of some cells through mitochondria damage. Arginine deprivation also induced the expression increase of p53 and p53AIP1, which aggravated cellular mitochondria damage. Chromatin autophagy appeared at the later stage of arginine deprivation. DNA damage came along with the whole process of arginine starvation. Histone 3 (H3) was found in autophagosomes, which implied that cancer cells try to utilize the arginine in histones to survive during arginine starvation.
Conclusions Mitochondria damage is the major mechanism for ADI expressed in cytosol to induce cancer cell death. Chromatophagy accumulation not only drives cancer cell to utilize histone arginine but also speeds up cancer cell death at the later time point of arginine deprivation.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
This is a list of supplementary files associated with this preprint. Click to download.
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