Gene expression of angiogenesis and apoptotic factors in female BALB/c mice with breast cancer after eight weeks of aerobic training

Document Type : Original Article

Authors

1 Department of Sport Sciences, Kish International Campus, University of Tehran, Kish, Iran

2 Department of Exercise Physiology, Faculty of Sport Sciences, University of Tehran, Tehran, Iran

Abstract

Objective(s): Breast cancer is the most common cancer in women, caused by a disorder in the angiogenesis and apoptosis process. Exercise can affect the process of angiogenesis and apoptosis in the tumor tissue. Thus, the aim of the present study was to investigate the changes in angiogenesis and apoptotic factors in mice with breast cancer after 8 weeks of exercise training.
Materials and Methods: Sixteen females BALB/c mice (age: 3-5 weeks and weight: 17.1 ± 0.1 g) with breast cancer were randomly divided into two groups of aerobic training and control. The aerobic training included 8 weeks and 5 sessions per week of running with an intensity of 14-20 m.min-1. HIF-1α, VEGF, miR-21 and cytochrome C, Apaf-1, caspase-9, and caspase-3 gene expressions were examined by real-time PCR. Repeated measures ANOVA, Bonferroni’s post hoc test, and independent samples t-test were used to analyze the data (p <0.05).
Results: The results showed that aerobic training reduced the growth of tumor volume and significantly reduced miR-21 gene expression. Aerobic training also significantly increased the gene expression of HIF-1α, cytochrome C, Apaf-1, caspase-9, and caspase-3, while changes in VEGF gene expression were not statistically significant.
Conclusion: It appears that aerobic exercise training reduces tumor size and ameliorates breast cancer by reducing miR-21 gene expression, suppressing the apoptosis process, and reducing angiogenesis.

Keywords


1. Thomadaki H, Scorilas A. Molecular profile of the BCL2 family of the apoptosis related genes in breast cancer cells after treatment with cytotoxic/cytostatic drugs. Connect Tissue Res 2008;49:261–264.
2. Venkatadri R, Muni T, Iyer AK, Yakisich JS, Azad N. Role of apoptosis-related miRNAs in resveratrol-induced breast cancer cell death. Cell Death Dis 2016;7:e2104.
3. Ribatti D. The discovery of antiangiogenic molecules: a historical review. Curr Pharm Des 2009;15:345-352.
4. Zhao D, Tu Y, Wan L, Bu L, Huang T, Sun X, et al. In Vivo monitoring of angiogenesis inhibition via down-regulation of mir-21 in a VEGFR2-Luc Murine breast cancer model using bioluminescent imaging. PLoS One 2013;8. e71472.
5. Pacher P, Beckman JS, Liaudet L. Nitric oxide and peroxynitrite in health and disease. Physiol Rev 2007;87:315–424.
6. García-Pastor C, Benito-Martínez S, Moreno-Manzano V, Fernández-Martínez AB, Lucio-Cazaña FJ. Mechanism and consequences of the impaired Hif-1α response to yypoxia in human proximal tubular HK-2 cells exposed to high glucose. Sci Rep 2019;9:15868.
7. Lv D, Cui PL, Yao SW, Xu YQ, Yang ZX. Melatonin inhibits the expression of vascular endothelial growth factor in pancreatic cancer cells. Chin J Cancer Res 2012;24:310-316.
8. Hassan M, Watari H, AbuAlmaaty A, Ohba Y, Sakuragi N. Apoptosis and molecular targeting therapy in cancer. Biomed Res Int 2014;2014:150845.
9. Glushakova OY, Glushakov AO, Borlongan CV, Valadka AB, Hayes RL, Glushakov AV. Role of caspase-3-mediated apoptosis in chronic caspase-3-cleaved tau accumulation and blood-brain barrier damage in the corpus callosum after traumatic brain injury in rats. J Neurotrauma 2018;35:157–173.
10. Green DR, Llambi F. Cell death signaling. Cold Spring Harb Perspect Biol 2015;7:a006080.
11. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007;35:495-516.
12. Shi M, Guo N. MicroRNA expression and its implications for the diagnosis and therapeutic strategies of breast cancer. Cancer Treat Rev 2009;35:328-334.
13. Lowery AJ, Miller N, McNeill RE, Kerin MJ. MicroRNAs as prognostic indicators and therapeutic targets: potential effect on breast cancer management. Clin Cancer Res 2008;14:360-365.
14. Si ML, Zhu S, Wu H, Lu Z, Wu F, Mo YY. MiR-21-mediated tumor growth. Oncogene 2007;26:2799-2803.
15. Wang N, Zhang CQ, He JH, Duan XF, Wang YY, Ji X, et al. MiR-21 down-regulation suppresses cell growth, invasion and induces cell apoptosis by targeting FASL, TIMP3, and RECK genes in esophageal carcinoma. Dig Dis Sci 2013;58:1863-1870.
16. Johnsson A, Johnsson A, Johansson K. Physical activity during and after adjuvant chemotherapy in patients with breast cancer. Physiotherapy 2013;99:221-227.
17. Hayes SC, Steele ML, Spence RR, Gordon L, Battistutta D, Bashford J, et al. Exercise following breast cancer: Exploratory survival analyses of two randomised, controlled trials. Breast Cancer Res Treat 2018;167:505–514.
18. Na HK, Oliynyk S. Effects of physical activity on cancer prevention. Ann N Y Acad Sci 2011;1229:176–183.
19. Egginton S. Invited review: activity-induced angiogenesis. Pflugers Arch 2009;457:963-977.
20. Nasiri M, Peeri M, Matinhomaei H. Endurance training attenuates angiogenesis following breast cancer by regulation of miR-126 and miR-296 in breast cancer bearing mice. Int J Cancer Manag 2017; 10:e8067.
21. Wahl P, Jansen F, Achtzehn S, Schmitz T, Bloch W, Mester J, et al. Effects of high intensity training and high volume training on endothelial microparticles and angiogenic growth factors. PLoS One 2014;9:e96024.
22. Faleiros CM, Francescato HD, Papoti M, Chaves L, Silva CG, Costa RS, et al. Effects of previous physical training on adriamycin nephropathy and its relationship with endothelial lesions and angiogenesis in the renal cortex. Life Sci 2017;169:43-51.
23. Khori V, Amani Shalamzari S, Isanejad A, Alizadeh AM, Alizadeh S, Khodayari S, et al. Effects of exercise training together with tamoxifen in reducing mammary tumor burden in mice: Possible underlying pathway of miR-21. Eur J Pharmacol 2015;765:179-187.
24. Jones LW, Viglianti BL, Tashjian JA, Kothadia SM, Keir ST, Freedland SJ, et al. Effect of aerobic exercise on tumor physiology in an animal model of human breast cancer. J Appl Physiol. 2010;108:343–348.
25. Ambros V. The functions of animal microRNAs. Nature 2004;431:350-355.
26. Myllyharju J, Koivunen P. Hypoxia-inducible factor prolyl 4-hydroxylases:common and specific roles. Biol Chem 2013;394:435–448.
27. Jones LW, Antonelli J, Masko EM, Broadwater G, Lascola CD, Fels D, et al. Exercise modulation of the host-tumor interaction in an orthotopic model of murine prostate cancer. J Appl Physiol 2012;113:263–272.
28. Wagner PD. Muscle intracellular oxygenation during exercise: optimization for oxygen transport, metabolism, and adaptive change. Eur J Appl Physiol 2012;112:1-8.
29. Hojman P, Dethlefsen C, Brandt C, Hansen J, Pedersen L, Pedersen BK. Exercise-induced muscle-derived cytokines inhibit mammary cancer cell growth. Am J Physiol - Endocrinol Metab 2011;301:504-510.
30. Liu LZ, Li C, Chen Q, Jing Y, Carpenter R, Jiang Y, et al. Mir-21 induced angiogenesis through AKT and ERK activation and HIF-1α expression. PLoS One 2011;6. e19139. 
31. Zhang Z, Yao L, Yang J, Wang Z, Du G. PI3K/Akt and HIF‑1 signaling pathway in hypoxia‑ischemia (Review). Mol Med Rep 2018;18:3547-3554.
32. Koyasu S, Kobayashi M, Goto Y, Hiraoka M, Harada H. Regulatory mechanisms of hypoxia-inducible factor 1 activity: Two decades of knowledge. Cancer Sci 2018;109:560-571.
33. Samaja M, Milano G. Adaptation to Hypoxia: A Chimera? Int J Mol Sci. 2020; 21: 1527.
34. Dufresne S, Rébillard A, Muti P, Friedenreich CM, Brenner DR. A Review of Physical Activity and Circulating miRNA Expression: Implications in Cancer Risk and Progression. Cancer Epidemiol Biomarkers Prev 2018;27:11-24.
35. Kuehbacher A, Urbich C, Dimmeler S. Targeting microRNA expression to regulate angiogenesis. Trends Pharmacol Sci 2008;29:12-15.
36. Baggish AL, Park J, Min PK, Isaacs S, Parker BA, Thompson PD, et al. Rapid upregulation and clearance of distinct circulating microRNAs after prolonged aerobic exercise. J Appl Physiol 2014;116:522–531.
37. Aoi W, Naito Y, Mizushima K, Takanami Y, Kawai Y, Ichikawa H, et al. The microRNA miR-696 regulates PGC-1α in mouse skeletal muscle in response to physical activity. Am J Physiol - Endocrinol Metab 2010; 298:E799-806.
38. Isanejad A, Alizadeh AM, Amani Shalamzari S, Khodayari H, Khodayari S, Khori V, et al. MicroRNA-206, let-7a and microRNA-21 pathways involved in the anti-angiogenesis effects of the interval exercise training and hormone therapy in breast cancer. Life Sci 2016;151:30-40.
39. Zimna A, Kurpisz M. Hypoxia-inducible factor-1 in physiological and pathophysiological angiogenesis: applications and therapies. Biomed Res Int. 2015;2015:549412.
40. Mehlem A, Palombo I, Wang X, Hagberg CE, Eriksson U, Falkevall A. PGC-1α coordinates mitochondrial respiratory capacity and muscular fatty acid uptake via regulation of VEGF-B. Diabetes 2016;65:861-873.
41. Shashar M, Chernichovski T, Pasvolsky O, Levi S, Grupper A, Hershkovitz R, et al. Vascular endothelial growth factor augments arginine transport and nitric oxide generation via a KDR receptor signaling pathway. Kidney Blood Press Res 2017;42:201-208.
42. Tolouei Azar J, Ravasi A, Soori R, Akbarnejad A, Hemati Nafar M. The Effect of 8 weeks aerobic training on angiogenesis (VEGF) and angiostatic (ES) factors In sedentary women. Stud Med Sci 2017;27:1032-1040.
43. Abotaleb M, Samuel SM, Varghese E, Varghese S, Kubatka P, Liskova A, et al. Flavonoids in cancer and cpoptosis. Cancers (Basel) 2018;11:28.
44. Indran IR, Tufo G, Pervaiz S, Brenner C. Recent advances in apoptosis, mitochondria and drug resistance in cancer cells. Biochim Biophys Acta 2011;1807:735-745.
45. Sen CK. Antioxidant and redox regulation of cellular signaling: introduction. Med Sci Sports Exerc 2001;33:368-370.
46. Buscaglia LE, Li Y. Apoptosis and the target genes of microRNA-21. Chin J Cancer 2011;30:371-380.
47. Medina PP, Nolde M, Slack FJ. OncomiR addiction in an in vivo model of microRNA-21-induced pre-B-cell lymphoma. Nature 2010;467:86-90.
48. Pfeffer CM, Singh ATK. Apoptosis: A target for anticancer therapy. Int J Mol Sci 2018;19:448-457.