Effects of pipecolic acid oxidase on proliferation, apoptosis, migration and invasion of primary liver cancer cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LÜ Yue , FENG Xuping , HUANG Tengda ,  YUAN Kefei ,  ZENG Yong

Laboratory of Liver Surgery, West China Hospital, Sichuan University, Chengdu 610041, P. R. China;

Corresponding author：

YUAN Kefei, Email: ykf13@163.com; ZENG Yong, Email: zengyong@medmail.com.cn

Keywords：

pipecolic acid oxidase; primary liver cancer; proliferation; apoptosis; migration and invasion

DOI：

10.7507/1007-9424.202405091

Video：

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Objective To investigate the effects of pipecolic acid oxidase (PIPOX) on the proliferation, apoptosis, migration and invasion of primary liver cancer cells. Methods Immunohistochemical staining and analysis of The Cancer Genome Atlas (TCGA) database were used to examine the PIPOX expression levels in liver cancer tissues and paired adjacent normal tissues, and studied their relationship with patient prognosis. Liver cancer cell lines stably overexpressing or knocking out PIPOX were constructed to explore PIPOX’s impact on liver cancer cell proliferation, apoptosis, migration and invasion by conducting in vitro functional experiments such as CCK-8, EdU, apoptosis detection, and Transwell assays. In vivo, nude mice subcutaneous tumor models and lung metastasis models were used to verify PIPOX’s effect on liver cancer growth and metastasis. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were both employed to detect the expression of epithelial-mesenchymal transition (EMT) markers in liver cancer cells. Results Immunohistochemical staining and TCGA database analysis revealed that PIPOX expression was significantly lower in liver cancer tissues compared to paired adjacent normal tissues (P<0.05). Prognostic analysis indicated shorter overall survival and disease-free survival in PIPOX low expression group (P<0.05). In vitro gain- and loss-of-function experiments showed that PIPOX significantly inhibited liver cancer cell migration and invasion (P<0.05), while having no significant effects on their proliferation and apoptosis (P>0.05). Animal experiments also confirmed that PIPOX significantly inhibited liver cancer lung metastasis (P<0.05), but had no significant effects on tumor growth (P>0.05). Finally, RT-qPCR and western blot results revealed that PIPOX promoted the expression of the epithelial marker E-cadherin (P<0.05) and inhibited the expression of mesenchymal markers (N-cadherin, vimentin, Snail) (P<0.05). Conclusions PIPOX significantly inhibits liver cancer cell migration and invasion, potentially via suppressing the EMT process. However, PIPOX does not significantly affect liver cancer cell proliferation and apoptosis.

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9.	Khan AP, Rajendiran TM, Ateeq B, et al. The role of sarcosine metabolism in prostate cancer progression. Neoplasia, 2013, 15(5): 491-501.
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18.	Zhang W, Sun HC, Wang WQ, et al. Sorafenib down-regulates expression of HTATIP2 to promote invasiveness and metastasis of orthotopic hepatocellular carcinoma tumors in mice. Gastroenterology, 2012, 143(6): 1641-1649.
19.	Pastushenko I, Blanpain C. EMT transition states during tumor progression and metastasis. Trends Cell Biol, 2019, 29(3): 212-226.
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21.	Giannelli G, Koudelkova P, Dituri F, et al. Role of epithelial to mesenchymal transition in hepatocellular carcinoma. J Hepatol, 2016, 65(4): 798-808.
22.	Lamouille S, Xu J, Derynck R. Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol, 2014, 15(3): 178-196.
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27.	Sreekumar A, Poisson LM, Rajendiran TM, et al. Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression. Nature, 2009, 457(7231): 910-914.

1. Bray F, Laversanne M, Sung H, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2024, 74(3): 229-263.
2. 郝运, 李川, 文天夫, 等. 全球及中国的肝癌流行病学特征: 基于《2022全球癌症统计报告》解读. 中国普外基础与临床杂志, 2024, 31(7): 781-789.
3. Li X, Ramadori P, Pfister D, et al. The immunological and metabolic landscape in primary and metastatic liver cancer. Nat Rev Cancer, 2021, 21(9): 541-557.
4. Ringelhan M, Pfister D, O’Connor T, et al. The immunology of hepatocellular carcinoma. Nat Immunol, 2018, 19(3): 222-232.
5. Sangro B, Sarobe P, Hervás-Stubbs S, et al. Advances in immunotherapy for hepatocellular carcinoma. Nat Rev Gastroenterol Hepatol, 2021, 18(8): 525-543.
6. IJlst L, de Kromme I, Oostheim W, et al. Molecular cloning and expression of human L-pipecolate oxidase. Biochem Biophys Res Commun, 2000, 270(3): 1101-1105.
7. Dodt G, Kim DG, Reimann SA, et al. L-Pipecolic acid oxidase, a human enzyme essential for the degradation of L-pipecolic acid, is most similar to the monomeric sarcosine oxidases. Biochem J, 2000, 345 Pt 3(Pt 3): 487-494.
8. Natarajan SK, Muthukrishnan E, Khalimonchuk O, et al. Evidence for pipecolate oxidase in mediating protection against hydrogen peroxide stress. J Cell Biochem, 2017, 118(7): 1678-1688.
9. Khan AP, Rajendiran TM, Ateeq B, et al. The role of sarcosine metabolism in prostate cancer progression. Neoplasia, 2013, 15(5): 491-501.
10. Yoon JK, Kim DH, Koo JS. Implications of differences in expression of sarcosine metabolism-related proteins according to the molecular subtype of breast cancer. J Transl Med, 2014, 12: 149. doi: 10.1186/1479-5876-12-149.
11. Kim HM, Lee YK, Koo JS. Expression of sarcosine-metabolizing enzymes in thyroid cancer. Int J Clin Exp Pathol, 2016, 9(7): 7132-7139.
12. Huang DQ, Singal AG, Kanwal F, et al. Hepatocellular carcinoma surveillance - utilization, barriers and the impact of changing aetiology. Nat Rev Gastroenterol Hepatol, 2023, 20(12): 797-809.
13. Llovet JM, Kelley RK, Villanueva A, et al. Hepatocellular carcinoma. Nat Rev Dis Primers, 2021, 7(1): 6. doi: 10.1038/s41572-020-00240-3.
14. 文天夫, 李川, 张晓赟, 等. “健康中国2030”肝癌5年生存率提高15%的关键. 中国普外基础与临床杂志, 2023, 30(11): 1281-1283.
15. Llovet JM, Pinyol R, Kelley RK, et al. Molecular pathogenesis and systemic therapies for hepatocellular carcinoma. Nat Cancer, 2022, 3(4): 386-401.
16. Yang X, Yang C, Zhang S, et al. Precision treatment in advanced hepatocellular carcinoma. Cancer Cell, 2024, 42(2): 180-197.
17. Forner A, Reig M, Bruix J. Hepatocellular carcinoma. Lancet, 2018, 391(10127): 1301-1314.
18. Zhang W, Sun HC, Wang WQ, et al. Sorafenib down-regulates expression of HTATIP2 to promote invasiveness and metastasis of orthotopic hepatocellular carcinoma tumors in mice. Gastroenterology, 2012, 143(6): 1641-1649.
19. Pastushenko I, Blanpain C. EMT transition states during tumor progression and metastasis. Trends Cell Biol, 2019, 29(3): 212-226.
20. Mittal V. Epithelial mesenchymal transition in tumor metastasis. Annu Rev Pathol, 2018, 13: 395-412.
21. Giannelli G, Koudelkova P, Dituri F, et al. Role of epithelial to mesenchymal transition in hepatocellular carcinoma. J Hepatol, 2016, 65(4): 798-808.
22. Lamouille S, Xu J, Derynck R. Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol, 2014, 15(3): 178-196.
23. Bracken CP, Goodall GJ. The many regulators of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol, 2022, 23(2): 89-90.
24. Dongre A, Weinberg RA. New insights into the mechanisms of epithelial-mesenchymal transition and implications for cancer. Nat Rev Mol Cell Biol, 2019, 20(2): 69-84.
25. Zheng X, Carstens JL, Kim J, et al. Epithelial-to-mesenchymal transition is dispensable for metastasis but induces chemoresistance in pancreatic cancer. Nature, 2015, 527(7579): 525-530.
26. Fischer KR, Durrans A, Lee S, et al. Epithelial-to-mesenchymal transition is not required for lung metastasis but contributes to chemoresistance. Nature, 2015, 527(7579): 472-476.
27. Sreekumar A, Poisson LM, Rajendiran TM, et al. Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression. Nature, 2009, 457(7231): 910-914.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Latest ArticlesEffects of pipecolic acid oxidase on proliferation, apoptosis, migration and invasion of primary liver cancer cells

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