Search JIM Advanced Search

Journal of Integrative Medicine ›› 2023, Vol. 21 ›› Issue (2): 184-193.doi: 10.1016/j.joim.2023.01.002

• Original Experimental Research • Previous Articles     Next Articles

Physical exercise suppresses hepatocellular carcinoma progression by alleviating hypoxia and attenuating cancer stemness through the Akt/GSK-3β/β-catenin pathway

Chu-lan Xiao a b 1, Zhi-peng Zhong a 1, Can Lü a 1, Bing-jie Guo a c, Jiao-jiao Chen a, Tong Zhao d, Zi-fei Yin e, Bai Li a   

  1. aDepartment of Rehabilitation, Changhai Hospital, Naval Medical University, Shanghai 200433, China
    bDepartment of Traditional Chinese Medicine, The 920th Hospital of Joint Logistics Support Force, Kunming 650000, Yunnan Province, China
    cSecond Team, Graduate School, Naval Medical University, Shanghai 200433, China
    dCollege of Integrated Traditional Chinese and Western Medicine, Nanjing University of Traditional Chinese Medicine, Nanjing 210023, Jiangsu Province, China
    eDepartment of Military Traditional Chinese Medicine, Faculty of Traditional Chinese Medicine, Naval Medical University, Shanghai 200433, China
  • Received:2022-05-18 Accepted:2022-10-27 Online:2023-03-20 Published:2023-04-06
  • Contact: Zi-fei Yin; Bai Li;

Physical exercise, a common non-drug intervention, is an important strategy in cancer treatment, including hepatocellular carcinoma (HCC). However, the mechanism remains largely unknown. Due to the importance of hypoxia and cancer stemness in the development of HCC, the present study investigated whether the anti-HCC effect of physical exercise is related to its suppression on hypoxia and cancer stemness.

A physical exercise intervention of swimming (30 min/d, 5 d/week, for 4 weeks) was administered to BALB/c nude mice bearing subcutaneous human HCC tumor. The anti-HCC effect of swimming was assessed in vivo by tumor weight monitoring, hematoxylin and eosin (HE) staining, and immunohistochemistry (IHC) detection of proliferating cell nuclear antigen (PCNA) and Ki67. The expression of stemness transcription factors, including Nanog homeobox (NANOG), octamer-binding transcription factor 4 (OCT-4), v-Myc avian myelocytomatosis viral oncogene homolog (C-MYC) and hypoxia-inducible factor-1α (HIF-1α), was detected using real-time reverse transcription polymerase chain reaction. A hypoxia probe was used to explore the intratumoral hypoxia status. Western blot was used to detect the expression of HIF-1α and proteins related to protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β)/β-catenin signaling pathway. The IHC analysis of platelet endothelial cell adhesion molecule-1 (CD31), and the immunofluorescence co-location of CD31 and desmin were used to analyze tumor blood perfusion. SMMC-7721 cells were treated with nude mice serum. The inhibition effect on cancer stemness in vitro was detected using suspension sphere experiments and the expression of stemness transcription factors. The hypoxia status was inferred by measuring the protein and mRNA levels of HIF-1α. Further, the expression of proteins related to Akt/GSK-3β/β-catenin signaling pathway was detected.

Swimming significantly reduced the body weight and tumor weight in nude mice bearing HCC tumor. HE staining and IHC results showed a lower necrotic area ratio as well as fewer PCNA or Ki67 positive cells in mice receiving the swimming intervention. Swimming potently alleviated the intratumoral hypoxia, attenuated the cancer stemness, and inhibited the Akt/GSK-3β/β-catenin signaling pathway. Additionally, the desmin+/CD31+ ratio, rather than the number of CD31+ vessels, was significantly increased in swimming-treated mice. In vitro experiments showed that treating cells with the serum from the swimming intervention mice significantly reduced the formation of SMMC-7721 cell suspension sphere, as well as the mRNA expression level of stemness transcription factors. Consistent with the in vivo results, HIF-1α and Akt/GSK-3β/β-catenin signaling pathway were also inhibited in cells treated with serum from swimming group.

Swimming alleviated hypoxia and attenuated cancer stemness in HCC, through suppression of the Akt/GSK-3β/β-catenin signaling pathway. The alleviation of intratumoral hypoxia was related to the increase in blood perfusion in the tumor.

Key words: Physical exercise, Swimming, Hepatocellular carcinoma, Liver cancer, Blood perfusion, Hypoxia, Cancer stemness, Akt/GSK-3β/β-catenin

[1] Cai-lian Fana, Wan-jun Caib, Meng-nan Ye, Miao Chen, Yi Dai. Qili Qiangxin, a compound herbal medicine formula, alleviates hypoxia-reoxygenation-induced apoptotic and autophagic cell death via suppression of ROS/AMPK/mTOR pathway in vitro. Journal of Integrative Medicine, 2022, 20(4): 365-375.
[2] Jia-jia Li, Qing Liang, Guang-chun Sun. Traditional Chinese medicine for prevention and treatment of hepatocellular carcinoma: A focus on epithelial-mesenchymal transition. Journal of Integrative Medicine, 2021, 19(6): 469-477.
[3] Gai Ran, Xi-lin Feng, Yi-lin Xie, Qing-yun Zheng, Peng-peng Guo, Ming Yang, Ying-lu Feng, Chen Ling, Li-qing Zhu, Chen Zhong. The use of miR122 and its target sequence in adeno-associated virus-mediated trichosanthin gene therapy. Journal of Integrative Medicine, 2021, 19(6): 515-525.
[4] Tong Zhao, Bing-jie Guo, Chu-lan Xiao, Jiao-jiao Chen, Can Lü, Fan-fu Fang, Bai Li. Aerobic exercise suppresses hepatocellular carcinoma by downregulating dynamin-related protein 1 through PI3K/AKT pathway. Journal of Integrative Medicine, 2021, 19(5): 418-427.
[5] Liang Ding, Xin-you Zhang, Di-yao Wu, Meng-ling Liu. Application of an extreme learning machine network with particle swarm optimization in syndrome classification of primary liver cancer. Journal of Integrative Medicine, 2021, 19(5): 395-407.
[6] Hao Gou, Ruo-chen Huang, Yong-hua Su, Wei Li. Design of dual targeting immunomicelles loaded with bufalin and study of their anti-tumor effect on liver cancer. Journal of Integrative Medicine, 2021, 19(5): 408-417.
[7] Kai-yue Tang, Shi-lin Du, Qian-lei Wang, Yi-fan Zhang, Hai-yan Song. Traditional chinese medicine targeting cancer stem cells as an alternative treatment for hepatocellular carcinoma. Journal of Integrative Medicine, 2020, 18(3): 196-202.
[8] Chang-quan Ling, Jia Fan, Hong-sheng Lin, Feng Shen, Zhen-ye Xu, Li-zhu Lin, Shu-kui Qin, Wei-ping Zhou, Xiao-feng Zhai, Bai Li, Qing-hui Zhou, on behalf of the Chinese Integrative Therapy of Primary Liver Cancer Working Group. Clinical practice guidelines for the treatment of primary liver cancer with integrative traditional Chinese and Western medicine. Journal of Integrative Medicine, 2018, 16(4): 236-248.
[9] Xuanbin Wang, Ning Wang, Fan Cheung, Lixing Lao, Charlie Li, Yibin Feng. Chinese medicines for prevention and treatment of human hepatocellular carcinoma: Current progress on pharmacological actions and mechanisms. Journal of Integrative Medicine, 2015, 13(3): 142-164.
[10] Dao-fang Ding, Xiao-feng Li, Hao Xu, Zhen Wang, Qian-qian Liang, Chen-guang Li, Yong-jun Wang. Mechanism of resveratrol on the promotion of induced pluripotent stem cells. Journal of Chinese Integrative Medicine, 2013, 11(6): 389-396.
[11] Xiao-feng Zhai, Zhe Chen, Bai Li, Feng Shen, Jia Fan, Wei-ping Zhou, Yun-ke Yang, Jing Xu, Xiao Qin, Le-qun Li, Chang-quan Ling. Traditional herbal medicine in preventing recurrence after resection of small hepatocellular carcinoma: A multicenter randomized controlled trial. Journal of Chinese Integrative Medicine, 2013, 11(2): 90-100.
[12] Daniel A. Weber, Janelle M. Wheat , Geoffrey M. Currie. Cancer stem cells and the impact of Chinese herbs, isolates and other complementary medical botanicals: A review. Journal of Chinese Integrative Medicine, 2012, 10(5): 493-503.
[13] Dong Fang-fang, Wang Yong-jun, Zhou Chong-jian. The relationship between hypoxia-inducible factor-1α and cartilage degeneration in osteoarthritis: a review. Journal of Chinese Integrative Medicine, 2012, 10(1): 13-18.
[14] Wang Yue-hua, He Xiao-li, Yang Hai-guang, Qin Hai-lin, Du Guan-hua. Effects of the active components of Chinese herbal medicine Xiaoxuming Decoction on memory behavior and brain injury in rats with chronic cerebral ischemia. Journal of Chinese Integrative Medicine, 2012, 10(1): 91-99.
[15] Xue-jun Du, Yan Lei , Jing Yang. Effects of Radix Ginseng and Radix Notoginseng formula on expressions of vascular endothelial growth factor receptor-2 and hypoxia-inducible factor-1α in ischemic myocardium of rats with acute myocardial infarction. Journal of Chinese Integrative Medicine, 2010, 8(6): 548-553.
Full text



[1] Dong Yang, Yong-ping Du, Qing Shen, Wei Chen, Yan Yu, Guang-lei Chen. Expression of alpha-smooth muscle actin in renal tubulointerstitium in patients with kidney collateral stasis. Journal of Chinese Integrative Medicine, 2008, 6(1): 41-44
[2] Xi Lin, Jian-ping Liu. Herbal medicines for viral myocarditis. Journal of Chinese Integrative Medicine, 2008, 6(1): 76
[3] Xi Lin, Jian-ping Liu. Tai chi for treating rheumatoid arthritis. Journal of Chinese Integrative Medicine, 2008, 6(1): 82
[4] Liang-ping Hu, Hui Gao. Discrimination of errors in statistical analysis of medical papers published in the first issue of 2006 in Journal of Chinese Integrative Medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 98-106
[5] Yan-bo Zhu , Qi Wang, Cheng-yu Wu, Guo-ming Pang, Jian-xiong Zhao, Shi-lin Shen, Zhong-yuan Xia , Xue Yan . Logistic regression analysis on relationships between traditional Chinese medicine constitutional types and overweight or obesity. Journal of Chinese Integrative Medicine, 2010, 8(11): 1023-1035
[6] Wei Xu, Meng Shi, Jian-gang Liu, Cheng-long Wang . Collagen protein expressions in ischemic myocardium of rats with acute myocardial infarction and effects of qi-tonifying, yin-tonifying and blood-activating herbs and detoxifying and blood-activating herbs. Journal of Chinese Integrative Medicine, 2010, 8(11): 1041-1047
[7] Tao Wang , Feng Qin. Effects of Chinese herbal medicine Xiaoyao Powder on monoamine neurotransmitters in hippocampus of rats with postpartum depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1075-1079
[8] Ying Xu , Chang-chun Zeng , Xiu-yu Cai , Rong-ping Guo , Guang Nie , Ying Jin. Chromaticity and optical spectrum colorimetry of the tongue color in different syndromes of primary hepatic carcinoma. Journal of Chinese Integrative Medicine, 2012, 10(11): 1263-1271
[9] Hai-feng Wei, Bai-liu Ya, Ling Zhao, Cui-fei Ye, Li Zhang, Lin Li. Evaluation of tongue manifestation of blood stasis syndrome and its relationship with blood rheological disorder in a rat model of transient brain ischemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 73-76
[10] Veena Sharma , Pracheta Janmeda , Ritu Paliwal , Shatruhan Sharma. Antihepatotoxic activity of Euphorbia neriifolia extract against N-nitrosodiethylamine-induced hepatocarcinogenesis in mice. Journal of Chinese Integrative Medicine, 2012, 10(11): 1303-1309