Significance of the Wnt canonical pathway in radiotoxicity via oxidative stress of electron beam radiation and its molecular control in mice

Abstract

Purpose

Radiation triggers cell death events through signaling proteins, but the combined mechanism of these events is unexplored The Wnt canonical pathway, on the other hand, is essential for cell regeneration and cell fate determination.

Aim

The relationship between the Wnt pathway's response to radiation and its role in radiotoxicity is overlooked, even though it is a critical molecular control of the cell. The Wnt pathway has been predicted to have radioprotective properties in some reports, but the overall mechanism is unknown. We intend to investigate how this combined cascade works throughout the radiation process and its significance over radiotoxicity.

Materials and methods

Thirty adult mice were irradiated with electron beam radiation, and 5 served as controls. Mice were sacrificed after 24 h and 30 days of irradiation. We assessed DNA damage studies, oxidative stress parameters, mRNA profiles, protein level (liver, kidney, spleen, and germ cells), sperm viability, and motility.

Observation

The mRNA profile helps to understand how the combined cascade of the Wnt pathway and NHEJ work together during radiation to combat oxidative response and cell survival. The quantitative examination of mRNA uncovers unique critical changes in all mRNA levels in all cases, particularly in germ cells. Recuperation was likewise seen in post-30 day’s radiation in the liver, spleen, and kidney followed by oxidative stress parameters, however not in germ cells. It proposes that reproductive physiology is exceptionally sensitive to radiation, even at the molecular level. It also suggests the suppression of Lef1/Axin2 could be the main reason for the permanent failure of the sperm function process. Post-irradiation likewise influences the morphology of sperm. The decrease in mRNA levels of Lef1, Axin2, Survivin, Ku70, and XRCC6 levels suggests radiation inhibits the Wnt canonical pathway and failure in DNA repair mechanisms in a coupled manner. An increase in Bax, Bcl2, and caspase3 suggests apoptosis activation followed by the decreased expression of enzymatic antioxidants.

Conclusion

Controlled several interlinked such as the Wnt canonical pathway, NHEJ pathway, and intrinsic apoptotic pathway execute when the whole body is exposed to radiation. These pathways decide the cell fate whether it will survive or will go to apoptosis which may further be used in a study to counterpart and better comprehend medication focus on radiation treatment

Graphical Abstract

(1) Electron beam radiation penetrates inside the cell leading to the production of free radicals, (2) which further produces ROS, RNS, and lipid peroxidation. (3) To counteract with ROS and RNS, SOD, Cat, and iNOS produce to scavenge free radicals. (4) Wnt3a, a natural activator of the Wnt canonical pathway binds to the Fz receptor and starts Wnt canonical pathway for cell proliferation and survival from radiation. (1) Electron beam radiation also cause DSB which further activates (5) NHEJ pathway for DNA repair. (1) Electron beam initiate the intrinsic apoptotic pathway (6) which leads to cell death. Survivin a candidate molecule of (4) Wnt canonical pathway inhibit the action of the intrinsic apoptotic pathway by inhibiting caspase 3 which rescue cell from apoptosis.

Keywords:

• Electron beam radiation
• Wnt canonical pathway
• apoptosis
• oxidative stress
• DNA damage
• radio-toxicity

Acknowledgments

The author wishes to thank Dr. Vishweshwara and HCG Bharath Cancer hospital staff for providing the irradiation facility. The author wishes sincere thanks to the Board of Research in Nuclear Sciences for providing us with non-interrupted funding to carry out this work.

Author contributions

Shashank Kumar conceived and designed the experiment, performed animal handling, harvesting of organs and germ cells, mRNA isolation, cDNA preparation, oxidative stress parameters, DNA ladder, Sperm morphology, viability, and motility. Shashank Kumar and Eram Fathima performed RT-qPCR analysis. Shashank Kumar performed the statistical analysis and prepared the manuscript. Suttur S. Malini and Farhath Khanum supervised the experiments. Suttur S. Malini corrected the manuscript. All authors read and approved the manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by the Board of Research in Nuclear Science, DAE, BARC (BRNS/2013/34/10429).

Notes on contributors

Shashank Kumar

Shashank Kumar, M.Sc. (Biotechnology) is working as Senior Research Fellow in DOS in Zoology, Manasagangothri, University of Mysore.

Eram Fathima

Eram Fathima, M.Sc. (Biotechnology) is working as Senior Research Fellow in NBT, Defense Food Research Laboratory, DRDO, Mysore.

Farhath Khanum

Farhath Khanum, Ph.D. is a retired Senior Scientist who worked as Scientist H in NBT, Defense Food Research Laboratory, DRDO, Mysore.

Suttur S. Malini

Suttur S. Malini, Ph.D. is Professor and Chairperson at DOS in Zoology and DOS in Genetics, University of Mysore, Mysore.