Long Non-Coding RNA ZEB2-AS1 Promotes Hepatocellular Carcinoma Progression by Regulating The miR-582-5p/FOXC1 Axis

Wu, Shimin; Chen, Juan; Liang, Ying; Luo, Qian; Tong, Yaoyao; Xie, Ling

doi:10.22074/cellj.2022.7963

Long Non-Coding RNA ZEB2-AS1 Promotes Hepatocellular Carcinoma Progression by Regulating The miR-582-5p/FOXC1 Axis

Document Type : Original Article

Authors

Center for Clinical Laboratory, General Hospital of The Yangtze River Shipping, Wuhan Brain Hospital, Wuhan, Hubei, China

10.22074/cellj.2022.7963

Abstract

Objective: Long non-coding RNAs (lncRNAs) feature prominently in tumors. Reportedly, lncRNA zinc finger E-box-binding homeobox 2 antisense RNA 1 (ZEB2-AS1) is aberrantly expressed in a variety of tumors. The present study was aimed to explore ZEB2-AS1 functions and determine mechanism in hepatocellular carcinoma
(HCC) progression.
Materials and Methods: In this experimental study, expressions of ZEB2-AS1, microRNA (miR)-582-5p and forkhead box C1 (FOXC1) mRNA in HCC tissues and cell lines were detected via quantitative reveres transcription polymerase chain reaction (qRT-PCR). After establishing gain- and loss-of-functions models, cell counting kit-8, 5-bromo-2’-deoxyuridine (BrdU), Transwell assays and flow cytometry analysis were conducted to examine HCC cell multiplication, migration, invasion and apoptosis, respectively. The targeted relationship between miR-582-5p and ZEB2-AS1 was verified via dual-luciferase reporter gene assay. Western blot was utilized for detecting FOXC1 expression in HCC cells after selectively regulating ZEB2-AS1 and miR-582-5p.
Results: In HCC tissues and cells, ZEB2-AS1 expression was increased. High ZEB2-AS1 expression was related to relatively large tumor volume, increased tumor-node-metastasis (TNM) stage and positive lymph node metastasis of the patients. ZEB2-AS1 overexpression facilitated HCC cell multiplication, migration, invasion and suppressed apoptosis, while ZEB2-AS1 knock-down caused the opposite effects. It was also confirmed that ZEB2-AS1 could competitively bind with miR-582-5p to repress its expression, and indirectly up-regulate FOXC1 expression level in HCC cells.
Conclusion: The current study revealed that ZEB2-AS1 was over-expressed in HCC tissues and cells. It also upregulated FOXC1, through sponging miR-582-5p, to promote HCC progression. This provides new perspectives for elucidating the pathogenesis of HCC.

Keywords

20.1001.1.22285806.2022.24.6.1.6

References

1. Wang D, Bai T, Chen G, Liu J, Chen M, Zhao Y, et al. Upregulation of long non-coding RNA FOXP4-AS1 and its regulatory network in hepatocellular carcinoma. Onco Targets Ther. 2019; 12: 7025-7038.
2. Liu J, Li W, Zhang J, Ma Z, Wu X, Tang L. Identification of key genes and long non-coding RNA associated ceRNA networks in hepatocellular carcinoma. PeerJ. 2019; 7: e8021.
3. Hartke J, Johnson M, Ghabril M. The diagnosis and treatment of hepatocellular carcinoma. Semin Diagn Pathol. 2017; 34(2): 153-159.
4. Tang X, Feng D, Li M, Zhou J, Li X, Zhao D, et al. Transcriptomic analysis of mRNA-lncRNA-miRNA interactions in hepatocellular carcinoma. Sci Rep. 2019; 9: 16096.
5. Tong MJ, Rosinski AA, Huynh CT, Raman SS, Lu DS. Long-term survival after surveillance and treatment in patients with chronic viral hepatitis and hepatocellular carcinoma. Hepatol Commun. 2017; 1(7): 595-608.
6. Su Y, Lv X, Yin W, Zhou L, Hu Y, Zhou A, et al. CircRNA Cdr1as functions as a competitive endogenous RNA to promote hepatocellular carcinoma progression. Aging (Albany NY). 2019; 11(9): 8182-8203.
7. Tiansheng G, Junming H, Xiaoyun W, Peixi C, Shaoshan D, Qianping C. LncRNA metastasis-associated lung adenocarcinoma transcript 1 promotes proliferation and invasion of non-Small cell lung cancer cells via down-regulating miR-202 expression. Cell J. 2020; 22(3): 375-385.
8. Quinn JJ, Chang HY. Unique features of long non-coding RNA biogenesis and function. Nat Rev Genet. 2016; 17(1): 47-62.
9. Peng WX, Koirala P, Mo YY. LncRNA-mediated regulation of cell signaling in cancer. Oncogene. 2017; 36(41): 5661-5667.
10. Liu F, Yuan JH, Huang JF, Yang F, Wang TT, Ma JZ, et al. Long noncoding RNA FTX inhibits hepatocellular carcinoma proliferation and metastasis by binding MCM2 and miR-374a. Oncogene. 2016; 35(41): 5422-5434.
11. Tang X, Zhang W, Ye Y, Li H, Cheng L, Zhang M, et al. LncRNA HOTAIR contributes to sorafenib resistance through suppressing miR-217 in hepatic carcinoma. Biomed Res Int. 2020; 2020: 9515071.
12. Kou JT, Ma J, Zhu JQ, Xu WL, Liu Z, Zhang XX, et al. LncRNA NEAT1 regulates proliferation, apoptosis and invasion of liver cancer. Eur Rev Med Pharmacol Sci. 2020; 24(8): 4152-4160.
13. Guo X, Jing YM, Lou HZ, Lou QA. Effect and mechanism of long non-coding RNA ZEB2-AS1 in the occurrence and development of colon cancer. Math Biosci Eng. 2019; 16(6): 8109–8120.
14. Lan T, Chang L, Wu L, Yuan Y. Downregulation of ZEB2-AS1 decreased tumor growth and metastasis in hepatocellular carcinoma. Mol Med Rep. 2016; 14(5): 4606-4612.
15. Stavast CJ, Erkeland SJ. The non-canonical aspects of microRNAs: many roads to gene regulation. Cells. 2019; 8(11): 1465.
16. Tian Y, Guan Y, Su Y, Luo W, Yang G, Zhang Y. MiR-582-5p inhibits bladder cancer-genesis by suppressing TTK expression. Cancer Manag Res. 2020; 12: 11933-11944.
17. Zhang Y, Huang W, Ran Y, Xiong Y, Zhong Z, Fan X, et al. miR- 582-5p inhibits proliferation of hepatocellular carcinoma by targeting CDK1 and AKT3. Tumour Biol. 2015; 36(11): 8309-8316.
18. Dai W, Meng X, Mo S, Xiang W, Xu Y, Zhang L, et al. FOXE1 represses cell proliferation and Warburg effect by inhibiting HK2 in colorectal cancer. Cell Commun Signal. 2020; 18(1): 7.
19. Lee YH, Lee HT, Chen CL, Chang CH, Hsu CY, Shyu WC. Role of FOXC1 in regulating APSCs self-renewal via STI-1/PrPC signaling. Theranostics. 2019; 9(22): 6443-6465.
20. Lin YJ, Shyu WC, Chang CW, Wang CC, Wu CP, Lee HT, et al. Tumor hypoxia regulates forkhead box C1 to promote lung cancer progression. Theranostics. 2017; 7(5): 1177-1191.
21. Huang W, Chen Z, Zhang L, Tian D, Wang D, Fan D, et al. Interleukin- 8 induces expression of FOXC1 to promote transactivation of CXCR1 and CCL2 in hepatocellular carcinoma cell lines and formation of metastases in mice. Gastroenterology. 2015; 149(4):1053-1067.
22. Wang WW, Chen B, Lei CB, Liu GX, Wang YG, Yi C, et al. miR- 582-5p inhibits invasion and migration of salivary adenoid cystic carcinoma cells by targeting FOXC1. Jpn J Clin Oncol. 2017; 47(8): 690-698.
23. Wang YL, Liu JY, Yang JE, Yu XM, Chen ZL, Chen YJ, et al. Lnc- UCID promotes G1/S transition and hepatoma growth by preventing DHX9-mediated CDK6 down-regulation. Hepatology. 2019; 70(1): 259-275.
24. Guo Y, Hu Y, Hu M, He J, Li B. Long non-coding RNA ZEB2-AS1 promotes proliferation and inhibits apoptosis in human lung cancer cells. Oncol Lett. 2018; 15(4): 5220-5226.
25. Qian Y, Wu X, Wang H, Hou G, Han X, Song W. MicroRNA-4290 suppresses PDK1-mediated glycolysis to enhance the sensitivity of gastric cancer cell to cisplatin. Braz J Med Biol Res. 2020; 53(5): e9330.
26. Huang S, Zou C, Tang Y, Wa Q, Peng X, Chen X, et al. miR-582-3p and miR-582-5p suppress prostate cancer metastasis to bone by repressing TGF-β signaling. Mol Ther Nucleic Acids. 2019; 16: 91-104.
27. Li L, Ma L. Upregulation of miR-582-5p regulates cell proliferation and apoptosis by targeting AKT3 in human endometrial carcinoma. Saudi J Biol Sci. 2018; 25(5): 965-970.
28. Gao H, Gong N, Ma Z, Miao X, Chen J, Cao Y, et al. LncRNA ZEB2-AS1 promotes pancreatic cancer cell growth and invasion through regulating the miR-204/HMGB1 axis. Int J Biol Macromol. 2018; 116: 545-551.
29. Xu Q, Liu H, Yu B, Chen W, Zhai L, Li X, et al. Long noncoding RNA ZEB2-AS1 facilitates laryngeal squamous cell carcinoma progression by miR-6840-3p/PLXNB1 axis. Onco Targets Ther. 2019; 12: 7337-7345.
30. Elian FA, Yan E, Walter MA. FOXC1, the new player in the cancer sandbox. Oncotarget. 2017; 9(8): 8165-8178.
31. Quintero-Ronderos P, Laissue P. The multisystemic functions of FOXD1 in development and disease. J Mol Med (Berl). 2018; 96(8): 725-739.
32. Yang Z, Jiang S, Cheng Y, Li T, Hu W, Ma Z, et al. FOXC1 in cancer development and therapy: deciphering its emerging and divergent roles. Ther Adv Med Oncol. 2017; 9(12): 797-816.
33. Bai X, Shao J, Zhou S, Zhao Z, Li F, Xiang R, et al. Inhibition of lung cancer growth and metastasis by DHA and its metabolite, RvD1, through miR-138-5p/FOXC1 pathway. J Exp Clin Cancer Res. 2019; 38(1): 479.
34. Hsu HH, Kuo WW, Shih HN, Cheng SF, Yang CK, Chen MC, et al. FOXC1 regulation of miR-31-5p confers oxaliplatin resistance by targeting LATS2 in colorectal cancer. Cancers (Basel). 2019; 11(10): 1576.