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Abstract
Folate and vitamin B12 deficiency is associated with depletion of the major intracellular antioxidant glutathione, and oxidative stress is emerging as an etiological mechanism for colon cancer. Azoxymethane (AOM), a potent carcinogen, induces colon cancer in rats by causing pathophysiological changes and oxidative stress. We investigated the synergistic effect of folate and vitamin B12 supplementation against AOM-induced carcinogenesis and oxidative stress in rat colon. Adult male rats were distributed into four groups: 1) Basal diet only; 2) AOM injection (15 mg/kg once per week in weeks 5 and 6); 3) Folate and vitamin B12 supplemented diet; 4) Folate and B12 diet with AOM injection. After 16 weeks, rats were sacrificed, colon tissue dissected, indicators of oxidative stress were measured, and immunohistochemical and ultrastructural changes were evaluated. AOM-injected rats showed oxidative stress, evident by glutathione depletion, oxidation of cellular proteins, and DNA oxidative damage. AOM increased mucosal levels of antiapoptotic and proapoptotic proteins Bcl2 and Bax and caused ultrastructure changes in colonic cell organelles. Folate and vitamin B12 supplementation decreased the level of oxidative stress and ameliorated the cytotoxic effects of AOM. In this in vivo experimental model of colon cancer, folate and vitamin B12 supplementation combats carcinogen-induced oxidative stress.
Acknowledgments
The authors acknowledge the assistance of Mr. Abdel Ghafar and Mr. Talal from the Histology Laboratory of Anatomy Department and Ms. Shadia from Immunohistochemistry Department for their dedicated effort in the immunohistochemistry experiments.
Disclosure Statement
No potential conflict of interest was reported by the authors.
Additional information
Funding
Notes on contributors
Smitha Padmanabhan
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Mostafa I. Waly
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Varna Taranikanti
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Nejib Guizani
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Amanat Ali
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Mohammad S. Rahman
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Zaher Al-Attabi
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Raya N. Al-Malky
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Sultan N. M. Al-Maskari
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Bader R. S. Al-Ruqaishi
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Jianan Dong
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.
Richard C. Deth
SP performed all the experiments. SP, MIW, and RCD conceived and wrote the first draft of the manuscript. JD and RCD jointly made critical revisions and arguments of the final manuscript. SP, VT, and MIW performed data analysis and prepared figures. VT, RNA, SNMA, and BRSA provided critical help in experiments and animal care. MIW, NG, AA, MSR, and ZA contributed in the experimental design, results interpretation, and review of literature. All authors have read and approved the final manuscript.