Effect of Swimming Exercise and Royal Jelly on MAPK and MMP-9 Gene Expression in Lung Tissue of Benzo[a]pyrene-Induced Lung Cancer Mouse Model
Subject Areas : Sport PhysiologySepideh Rajabi Baniani 1 , Seyed Ali Hosseini 2 , Mehrzad Moghadasi 3
1 - PhD Student, Department of sport sciences, Sh.C., Islamic Azad University, Shiraz, Iran
2 - Department of sport sciences, Marv.C., Islamic Azad University, Marvdasht, Iran
3 - Department of sport sciences, Sh.C., Islamic Azad University, Shiraz, Iran
Keywords: : Lung cancer, Mitogen-Activated Protein Kinase, Matrix Metalloproteinase-9, Royal Jelly, Swimming Exercise,
Abstract :
Background and Objectives: Lung cancer is one of the most prevalent cancers with a high mortality rate. Mitogen-Activated Protein Kinase (MAPK) and (Matrix Metalloproteinase-9) MMP-9 pathways play crucial roles in inflammation, oxidative stress, and cellular invasion. This study aimed to investigate the effects of swimming exercise and royal jelly on the gene expression of MAPK and MMP-9 in the lung tissue of mice with benzo[a]pyrene-induced lung cancer.
Materials and Methods: In this experimental study, 48 male Balb/C mice aged eight weeks were divided into eight groups: healthy control, lung cancer (BZP), , lung cancer + swimming training (ST), , lung cancer + royal jelly 50 mg/kg (RJ50), , lung cancer + royal jelly 100 mg/kg (RJ100), , lung cancer + swimming + royal jelly 50 mg/kg (ST.RJ50), and , lung cancer + swimming + royal jelly 100 mg/kg (ST.RJ100). Lung cancer was induced by intraperitoneal injection of benzo[a]pyrene at 100 mg/kg body weight. Swimming training was performed 3 days per week for 12 weeks. Royal jelly was administered intraperitoneally at doses of 50 and 100 mg/kg body weight. Forty-eight hours after the last session, mice were sacrificed, lung tissues were harvested, and gene expression of MAPK and MMP-9 was measured using real-time PCR.
Results: Lung cancer induction significantly increased MAPK and MMP-9 expression. Individual interventions, including swimming and royal jelly at 100 mg/kg, partially reduced MAPK and MMP-9 expression. However, the combination of swimming exercise with royal jelly, particularly at the higher dose (ST.RJ100), produced the greatest reduction in the expression of both genes.
Conclusion: The combination of regular exercise with royal jelly, especially at higher doses, has a synergistic effect in reducing MAPK pathway activity and MMP-9 expression. This combination may serve as a preventive and complementary natural strategy to control lung cancer progression and invasion. These findings highlight the importance of simultaneous non-pharmacological interventions with antioxidant and anti-inflammatory effects to improve molecular pathways related to cancer and enhance disease management.
1. Zhao M, Xue G, He B, Deng J, Wang T, Zhong Y, et al. Integrated multiomics signatures to optimize the accurate diagnosis of lung cancer. Nature Communications. 2025;16(1):84.
2. Smolarz B, Łukasiewicz H, Samulak D, Piekarska E, Kołaciński R, Romanowicz H. Lung Cancer—Epidemiology, Pathogenesis, Treatment and Molecular Aspect (Review of Literature). International Journal of Molecular Sciences. 2025;26(5):2049.
3. Naghizadeh, N., Riyahi Malayeri, S., Roozbahani, M., Khademi, A., Shirvani, H. The Effectiveness of Endurance Training and Nano Curcumin Supplementation on the Expression of Mir-21and P53 Genes in Brain Tumor Tissue in an Animal Model of Glioblastoma Multiform. Journal of Nutrition,Fasting and Health, 2024; 12(1): 26-35. doi: 10.22038/jnfh.2023.73964.1455.
4. Xie X, Shin YR, Kim T-S, Seong Y-S, Yi YW, Kim DJ. Identification of sinensetin as a selective inhibitor for mitogen-activated protein kinase kinase 6 and an anticancer agent for non-small cell lung cancer. American Journal of Cancer Research. 2025;15(1):113.
5. Alsharoh H, Chiroi P, Isachesku E, Tanasa RA, Pop O-L, Pirlog R, Berindan-Neagoe I. Personalizing therapy outcomes through mitogen-activated protein kinase pathway inhibition in non-small cell lung cancer. Biomedicines. 2024;12(7):1489.
6. Atkinson JJ, Senior RM. Matrix metalloproteinase-9 in lung remodeling. American journal of respiratory cell and molecular biology. 2003;28(1):12-24.
7. Chen L-H, Tsai Y-F, Wu W-T, Chiu K-L, Tsai C-W, Chang W-S, et al. Association of matrix metalloproteinase-9 genotypes with lung cancer risk in Taiwan. Anticancer Research. 2024;44(5):1845-52.
8. Gao L, Dhilipkannah P, Jiang F. Streptococcus pneumoniae infection is associated with matrix metalloproteinase-9 in lung cancer progression and brain metastases. Scientific Reports. 2025;15(1):37007.
9. Zhang H, Zhao B, Zhai Z-G, Zheng J-D, Wang Y-K, Zhao Y-Y. Expression and clinical significance of MMP-9 and P53 in lung cancer. European Review for Medical & Pharmacological Sciences. 2021;25(3).
10. Kalali D. The role of the matrix metalloproteinase-9 gene in tumor development and metastasis: a narrative review. Global Medical Genetics. 2023;10(02):048-53.
11. Marino P, Mininni M, Deiana G, Marino G, Divella R, Bochicchio I, et al. Healthy lifestyle and cancer risk: modifiable risk factors to prevent cancer. Nutrients. 2024;16(6):800.
12. Porro C, La Torre ME, Tartaglia N, Benameur T, Santini M, Ambrosi A, et al. The potential role of nutrition in lung cancer establishment and progression. Life. 2022;12(2):270.
13. Yang L, Morielli AR, Heer E, Kirkham AA, Cheung WY, Usmani N, et al. Effects of exercise on cancer treatment efficacy: a systematic review of preclinical and clinical studies. Cancer research. 2021;81(19):4889-95.
14. Luo Z, Wan R, Liu S, Feng X, Peng Z, Wang Q, et al. Mechanisms of exercise in the treatment of lung cancer–a mini-review. Frontiers in Immunology. 2023;14:1244764.
15. SH M-H, Musavi N. The effect of 12 weeks of submaximal swimming training on immunoreactivity of Ras and Raf-1 in lung epithelial cells of Wistar rats exposed to carcinogen NNK. 2020.
16. Williamson D, Gallagher P, Harber M, Hollon C, Trappe S. Mitogen‐activated protein kinase (MAPK) pathway activation: effects of age and acute exercise on human skeletal muscle. The Journal of physiology. 2003;547(3):977-87.
17. Gibala MJ, McGee SL, Garnham AP, Howlett KF, Snow RJ, Hargreaves M. Brief intense interval exercise activates AMPK and p38 MAPK signaling and increases the expression of PGC-1α in human skeletal muscle. Journal of applied physiology. 2009;106(3):929-34.
18. Messina G, Tartaglia N, Ambrosi A, Porro C, Campanozzi A, Valenzano A, et al. The beneficial effects of physical activity in lung cancer prevention and/or treatment. Life. 2022;12(6):782.
19. Ababd ZVM, Chaman NH, Hosseini M, Maleki A. Aerobic Exercise and Nano-curcumin Supplementation Prevent cancer symptom development through MAPK/ERK pathway. 2023.
20. Rullman E, Olsson K, Wågsäter D, Gustafsson T. Circulating MMP-9 during exercise in humans. European journal of applied physiology. 2013;113(5):1249-55.
21. Giganti MG, Tresoldi I, Sorge R, Melchiorri G, Triossi T, Masuelli L, et al. Physical exercise modulates the level of serum MMP-2 and MMP-9 in patients with breast cancer. Oncology letters. 2016;12(3):2119-26.
22. Oršolić N, Jazvinšćak Jembrek M. Royal jelly: biological action and health benefits. International journal of molecular sciences. 2024;25(11):6023.
23. Miyata Y, Sakai H. Anti-cancer and protective effects of royal jelly for therapy-induced toxicities in malignancies. International journal of molecular sciences. 2018;19(10):3270.
24. Kul Köprülü T, Gezer B, Erkal Çam B. The impact of apoptosis-inducing MAPK and glycolytic pathways modulated by Aloe vera and royal jelly in lung and colorectal cancer. Medical Oncology. 2025;42(2):51.
25. Fazili N, Soheili ZS, Malekzadeh-Shafaroudi S, Samiei S, Alipoor SD, Moshtaghi N, Bagheri A. Royal Jelly Decreases MMP-9 Expression and Induces Apoptosis in Human 5637 Bladder Cancer Cells. Journal of Cell & Molecular Research. 2021;13(1).
26. Rashedi F, Heidarnia E, Hosseini SA. Swimming training and royal jelly effects on caspase-3 expression in lung cancer mice. Journal of Physical Activity and Hormones. 2025;6(1):14-8.
27. Fei Z, Li D, Li K, Zhou M, Li Y, Li Y, Sun Z. Detraining after tumor-bearing accelerates tumor growth while continuous training decreases tumor growth in mice. Journal of Traditional Chinese Medical Sciences. 2020;7(1):75-81.
28. Mirdar S, Arab A, Hedayati M, Hajizade A. The effect of pregnant rat swimming on hypoxia-inducible factor-1α levels of neonatal lung. Tehran University Medical Journal. 2012;69(12).
29. Shirzad M, Kordyazdi R, Shahinfard N, Nikokar M. Does Royal jelly affect tumor cells? Journal of HerbMed Pharmacology. 2013;2(2):45-8.
30. Yao J, Xiong S, Klos K, Nguyen N, Grijalva R, Li P, Yu D. Multiple signaling pathways involved in activation of matrix metalloproteinase-9 (MMP-9) by heregulin-β1 in human breast cancer cells. Oncogene. 2001;20(56):8066-74.
31. Cho SJ, Chae MJ, Shin BK, Kim HK, Kim A. Akt-and MAPK-mediated activation and secretion of MMP-9 into stroma in breast cancer cells upon heregulin treatment. Molecular medicine reports. 2008;1(1):83-8.
32. Liu A, Li X, Hao Z, Cao J, Li H, Sun M, et al. Alterations of DNA methylation and mRNA levels of CYP1A1, GSTP1, and GSTM1 in human bronchial epithelial cells induced by benzo [a] pyrene. Toxicology and Industrial Health. 2022;38(3):127-38.
33. Radak Z, Zhao Z, Koltai E, Ohno H, Atalay M. Oxygen consumption and usage during physical exercise: the balance between oxidative stress and ROS-dependent adaptive signaling. Antioxidants & redox signaling. 2013;18(10):1208-46.
34. Fashi M, Alinejad HA, Mahabadi HA. The effect of aerobic exercise in ambient particulate matter on lung tissue inflammation and lung cancer. Iranian journal of cancer prevention. 2015;8(3):e2333.
35. Du T, Wang W, Zhang R. Royal jelly and doxorubicin suppressed tumor cells in the xenograft model of lung cancer via the STAT3/FOXM1/ATG7 signaling pathways in athymic nude mice: a biochemical, immunohistochemically and molecular approach. Toxicology Research. 2025;14(2):tfaf042.
36. Aslan A, Gok O, Beyaz S, Can MI, Parlak G, Gundogdu R, et al. Royal jelly regulates the caspase, Bax and COX-2, TNF-α protein pathways in the fluoride exposed lung damage in rats. Tissue and Cell. 2022;76:101754.
37. Mohammadi H, Mohammadian B, Najafzadeh Varzi H, Shahriari A. Evaluation of the effect of royal jelly on histopathological and biochemical changes in the lungs of rats following arsenic trioxide toxicity. Iranian Veterinary Journal. 2024;20(2):92-101.
38. Benoot T, Piccioni E, De Ridder K, Goyvaerts C. TNFα and immune checkpoint inhibition: friend or foe for lung cancer? International Journal of Molecular Sciences. 2021;22(16):8691.
39. Dong Y, Sun N, Qiang Y, Wang Y, Yuan Y, Li M. TNF-α inhibites non-small cell lung cancer cells proliferation by targeting THRIL in a FTO-YTHDF2-dependent manner. Archives of Biochemistry and Biophysics. 2025:110438.
40. Sobo-Vujanovic A, Vujanovic L, DeLeo AB, Concha-Benavente F, Ferris RL, Lin Y, Vujanovic NL. Inhibition of soluble tumor necrosis factor prevents chemically induced carcinogenesis in mice. Cancer immunology research. 2016;4(5):441-51.
41. Alim LF, Keane C, Souza-Fonseca-Guimaraes F. Molecular mechanisms of tumour necrosis factor signalling via TNF receptor 1 and TNF receptor 2 in the tumour microenvironment. Current opinion in immunology. 2024;86:102409.
42. Riyahi Malayeri S, Hoseini M. The Effect of Acute Exercise on Vaspin and Chemerin Levels in Obese Men. jbrms 2021; 8 (1) :58-66
43. Torrey H, Butterworth J, Mera T, Okubo Y, Wang L, Baum D, et al. Targeting TNFR2 with antagonistic antibodies inhibits proliferation of ovarian cancer cells and tumor-associated Tregs. Science signaling. 2017;10(462):eaaf8608.
44. Tam EM, Fulton RB, Sampson JF, Muda M, Camblin A, Richards J, et al. Antibody-mediated targeting of TNFR2 activates CD8+ T cells in mice and promotes antitumor immunity. Science translational medicine. 2019;11(512):eaax0720.
