FXR1 promotes proliferation, invasion and migration of hepatocellular carcinoma <em>in vitro</em> and <em>in vivo</em>

Zhao,Kun; Gao,Jie; Shi,Jihua; Shi,Chengcheng; Pang,Chun; Li,Jie; Guo,Wenzhi; Zhang,Shuijun

doi:10.3892/ol.2022.13608

FXR1 promotes proliferation, invasion and migration of hepatocellular carcinoma in vitro and in vivo

Authors:
- Kun Zhao
- Jie Gao
- Jihua Shi
- Chengcheng Shi
- Chun Pang
- Jie Li
- Wenzhi Guo
- Shuijun Zhang
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Affiliations: Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China, Department of Pharmacy, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China
Published online on: November 22, 2022 https://doi.org/10.3892/ol.2022.13608
Article Number: 22
Copyright: © Zhao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hepatocellular carcinoma (HCC) is a common malignancy that is associated with a poor prognosis. The extensively studied TGF‑β pathway is mediated by SMAD proteins. FXR1, a protein‑coding gene belonging to the fragile X‑related (FXR) family, is involved in the TGF‑β pathway. Previous studies have shown that FXR1 promotes the proliferation, invasion, and migration of colorectal cancer cells. The aim of the present study was to explore the effects of FXR1 on HCC via the TGF‑β/SMAD signaling pathway. Immunohistochemical analysis was used to detect the expression of FXR1 in HCC and normal tissues. Western blotting was used to detect protein expression levels in the HCC cell lines, cell migration and invasion were assessed using Transwell assays, and cell proliferation was assessed using a colony formation assay. The ability of the liver cancer cells to grow in vivo was investigated using a nude mouse tumor‑bearing model. The results showed that FXR1 expression was upregulated in HCC tissues compared with normal tissues. Knockdown of FXR1 resulted in reduced expression of SMAD2/3 and EMT‑related proteins in HCC cells. In addition, FXR1 knockdown inhibited the proliferation, migration, and invasion of HCC cells. FXR1 knockdown also reversed the promoting effect of TGF‑β on the invasive ability of HCC cells. Knockdown of SMAD2/3 reversed the increase in HCC cell invasion induced by FXR1 overexpression. Finally, upregulated FXR1 expression was associated with a poorer prognosis in patients with HCC. In conclusion, FXR1 promoted HCC proliferation, migration, and invasion through the regulation of SMAD2/3.

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1	Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A and Bray F: Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 71:209–249. 2021. View Article : Google Scholar : PubMed/NCBI
2	Xie Y: Hepatitis B virus-associated hepatocellular carcinoma. Adv Exp Med Biol. 1018:11–21. 2017. View Article : Google Scholar : PubMed/NCBI
3	Jiang Y, Han QJ and Zhang J: Hepatocellular carcinoma: Mechanisms of progression and immunotherapy. World J Gastroenterol. 25:3151–3167. 2019. View Article : Google Scholar : PubMed/NCBI
4	Piñero F, Dirchwolf M and Pessôa MG: Biomarkers in hepatocellular carcinoma: Diagnosis, prognosis and treatment response assessment. Cells. 9:13702020. View Article : Google Scholar : PubMed/NCBI
5	Dimri M and Satyanarayana A: Molecular signaling pathways and therapeutic targets in hepatocellular carcinoma. Cancers (Basel). 12:4912020. View Article : Google Scholar : PubMed/NCBI
6	Akula SM, Abrams SL, Steelman LS, Emma MR, Augello G, Cusimano A, Azzolina A, Montalto G, Cervello M and McCubrey JA: RAS/RAF/MEK/ERK, PI3K/PTEN/AKT/mTORC1 and TP53 pathways and regulatory miRs as therapeutic targets in hepatocellular carcinoma. Expert Opin Ther Targets. 23:915–929. 2019. View Article : Google Scholar : PubMed/NCBI
7	Fujiwara N, Friedman SL, Goossens N and Hoshida Y: Risk factors and prevention of hepatocellular carcinoma in the era of precision medicine. J Hepatol. 68:526–549. 2018. View Article : Google Scholar : PubMed/NCBI
8	Xing R, Gao J, Cui Q and Wang Q: Strategies to improve the antitumor effect of immunotherapy for hepatocellular carcinoma. Front Immunol. 12:7832362021. View Article : Google Scholar : PubMed/NCBI
9	Chen Y, Li L, Lan J, Cui Y, Rao X, Zhao J, Xing T, Ju G, Song G, Lou J and Liang J: CRISPR screens uncover protective effect of PSTK as a regulator of chemotherapy-induced ferroptosis in hepatocellular carcinoma. Mol Cancer. 21:112022. View Article : Google Scholar : PubMed/NCBI
10	Hoogeveen AT, Willemsen R and Oostra BA: Fragile X syndrome, the Fragile X related proteins, and animal models. Micros Res Tech. 57:148–155. 2002. View Article : Google Scholar : PubMed/NCBI
11	Majumder M, Johnson RH and Palanisamy V: Fragile X-related protein family: A double-edged sword in neurodevelopmental disorders and cancer. Crit Rev Biochem Mol Biol. 55:409–424. 2020. View Article : Google Scholar : PubMed/NCBI
12	Jin X, Zhai B, Fang T, Guo X and Xu L: FXR1 is elevated in colorectal cancer and acts as an oncogene. Tumour Biol. 37:2683–2690. 2016. View Article : Google Scholar : PubMed/NCBI
13	Majumder M and Palanisamy V: RNA binding protein FXR1-miR301a-3p axis contributes to p21WAF1 degradation in oral cancer. PLoS Genet. 16:e10085802020. View Article : Google Scholar : PubMed/NCBI
14	Cao S, Zheng J, Liu X, Liu Y, Ruan X, Ma J, Liu L, Wang D, Yang C, Cai H, et al: FXR1 promotes the malignant biological behavior of glioma cells via stabilizing MIR17HG. J Exp Clin Cancer Res. 38:372019. View Article : Google Scholar : PubMed/NCBI
15	Zhao M, Mishra L and Deng CX: The role of TGF-β/SMAD4 signaling in cancer. Int J Biol Sci. 14:111–123. 2018. View Article : Google Scholar : PubMed/NCBI
16	Colak S and Ten Dijke P: Targeting TGF-β signaling in cancer. Trends Cancer. 3:56–71. 2017. View Article : Google Scholar : PubMed/NCBI
17	Hao Y, Baker D and Ten Dijke P: TGF-β-mediated epithelial-mesenchymal transition and cancer metastasis. Int J Mol Sci. 20:27672019. View Article : Google Scholar : PubMed/NCBI
18	Hu HH, Chen DQ, Wang YN, Feng YL, Cao G, Vaziri ND and Zhao YY: New insights into TGF-β/Smad signaling in tissue fibrosis. Chem Biol Interact. 292:76–83. 2018. View Article : Google Scholar : PubMed/NCBI
19	Aashaq S, Batool A, Mir SA, Beigh MA, Andrabi KI and Shah ZA: TGF-β signaling: A recap of SMAD-independent and SMAD-dependent pathways. J Cell Physiol. 237:59–85. 2021. View Article : Google Scholar : PubMed/NCBI
20	Pallasch FB and Schumacher U: Angiotensin Inhibition, TGF-β and EMT in Cancer. Cancers (Basel). 12:27852020. View Article : Google Scholar : PubMed/NCBI
21	Suarez-Carmona M, Lesage J, Cataldo D and Gilles C: EMT and inflammation: Inseparable actors of cancer progression. Mol Oncol. 11:805–823. 2017. View Article : Google Scholar : PubMed/NCBI
22	Babaei G, Aziz SG and Jaghi NZZ: EMT, cancer stem cells and autophagy; The three main axes of metastasis. Biomed Pharmacother. 133:1109092021. View Article : Google Scholar : PubMed/NCBI
23	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
24	Syed V: TGF-β signaling in cancer. J Cell Biochem. 117:1279–1287. 2016. View Article : Google Scholar : PubMed/NCBI
25	Zhang K, Fang T, Shao Y and Wu Y: TGF-β-MTA1-SMAD7-SMAD3-SOX4-EZH2 signaling axis promotes viability, migration, invasion and EMT of hepatocellular carcinoma cells. Cancer Manag Res. 13:7087–7099. 2021. View Article : Google Scholar : PubMed/NCBI
26	Cao H, Gao R, Yu C, Chen L and Feng Y: The RNA-binding protein FXR1 modulates prostate cancer progression by regulating FBXO4. Funct Integr Genomics. 19:487–496. 2019. View Article : Google Scholar : PubMed/NCBI
27	Li K, Du Y, Li L and Wei DQ: Bioinformatics approaches for anti-cancer drug discovery. Curr Drug Targets. 21:3–17. 2020. View Article : Google Scholar : PubMed/NCBI
28	Anashkina AA, Leberfarb EY and Orlov YL: Recent trends in cancer genomics and bioinformatics tools development. Int J Mol Sci. 22:121462021. View Article : Google Scholar : PubMed/NCBI
29	Tsimberidou AM: Targeted therapy in cancer. Cancer Chemother Pharmacol. 76:1113–1132. 2015. View Article : Google Scholar : PubMed/NCBI
30	Yoshida K, Murata M, Yamaguchi T, Matsuzaki K and Okazaki K: Reversible human TGF-β signal shifting between tumor suppression and fibro-carcinogenesis: Implications of smad phospho-isoforms for hepatic epithelial-mesenchymal transitions. J Clin Med. 5:72016. View Article : Google Scholar : PubMed/NCBI
31	Fransvea E, Angelotti U, Antonaci S and Giannelli G: Blocking transforming growth factor-beta up-regulates E-cadherin and reduces migration and invasion of hepatocellular carcinoma cells. Hepatology. 47:1557–1566. 2008. View Article : Google Scholar : PubMed/NCBI
32	Bai X, Yi M, Jiao Y, Chu Q and Wu K: Blocking TGF-β signaling to enhance the efficacy of immune checkpoint inhibitor. Onco Targets Ther. 12:9527–9538. 2019. View Article : Google Scholar : PubMed/NCBI
33	Pastushenko I and Blanpain C: EMT transition states during tumor progression and metastasis. Trends Cell Biol. 29:212–226. 2019. View Article : Google Scholar : PubMed/NCBI
34	Bakir B, Chiarella AM, Pitarresi JR and Rustgi AK: EMT, MET, plasticity, and tumor metastasis. Trends Cell Biol. 30:764–776. 2020. View Article : Google Scholar : PubMed/NCBI
35	Lu W and Kang Y: Epithelial-mesenchymal plasticity in cancer progression and metastasis. Dev Cell. 49:361–374. 2019. View Article : Google Scholar : PubMed/NCBI
36	Yuan GJ, Li QW, Shan SL, Wang WM, Jiang S and Xu XM: Hyperthermia inhibits hypoxia-induced epithelial-mesenchymal transition in HepG2 hepatocellular carcinoma cells. World J Gastroenterol. 18:4781–4786. 2012. View Article : Google Scholar : PubMed/NCBI
37	Kimura-Tsuchiya R, Ishikawa T, Kokura S, Mizushima K, Adachi S, Okajima M, Matsuyama T, Okayama T, Sakamoto N, Katada K, et al: The inhibitory effect of heat treatment against epithelial-mesenchymal transition (EMT) in human pancreatic adenocarcinoma cell lines. J Clin Biochem Nutr. 55:56–61. 2014. View Article : Google Scholar : PubMed/NCBI
38	Wang J, Xu Z, Wang Z, Du G and Lun L: TGF-beta signaling in cancer radiotherapy. Cytokine. 148:1557092021. View Article : Google Scholar : PubMed/NCBI
39	Zhang M, Zhang YY, Chen Y, Wang J, Wang Q and Lu H: TGF-β signaling and resistance to cancer therapy. Front Cell Dev Biol. 9:7867282021. View Article : Google Scholar : PubMed/NCBI
40	Li H, He G, Yao H, Song L, Zeng L, Peng X, Rosol TJ and Deng X: TGF-β induces degradation of PTHrP through ubiquitin-proteasome system in hepatocellular carcinoma. J Cancer. 6:511–518. 2015. View Article : Google Scholar : PubMed/NCBI
41	Suwa K, Yamaguchi T, Yoshida K, Murata M, Ichimura M, Tsuneyama K, Seki T and Okazaki K: Smad phospho-isoforms for hepatocellular carcinoma risk assessment in patients with nonalcoholic steatohepatitis. Cancers (Basel). 12:2862020. View Article : Google Scholar : PubMed/NCBI
42	Yoshida K, Matsuzaki K, Murata M, Yamaguchi T, Suwa K and Okazaki K: Clinico-pathological importance of TGF-β/phospho-smad signaling during human hepatic fibrocarcinogenesis. Cancers (Basel). 10:1832018. View Article : Google Scholar : PubMed/NCBI
43	Kaminska B, Wesolowska A and Danilkiewicz M: TGF beta signalling and its role in tumour pathogenesis. Acta Biochim Pol. 52:329–337. 2005. View Article : Google Scholar : PubMed/NCBI
44	Burkhart RA, Ronnekleiv-Kelly SM and Pawlik TM: Personalized therapy in hepatocellular carcinoma: Molecular markers of prognosis and therapeutic response. Surg Oncol. 26:138–145. 2017. View Article : Google Scholar : PubMed/NCBI
45	Qin LX and Tang ZY: The prognostic molecular markers in hepatocellular carcinoma. World J Gastroenterol. 8:385–392. 2002. View Article : Google Scholar : PubMed/NCBI
46	Stefani C, Miricescu D, Stanescu S II, Nica RI, Greabu M, Totan AR and Jinga M: Growth factors, PI3K/AKT/mTOR and MAPK signaling pathways in colorectal cancer pathogenesis: Where are we now? Int J Mol Sci. 22:102602021. View Article : Google Scholar : PubMed/NCBI
47	Shorning BY, Dass MS, Smalley MJ and Pearson HB: The PI3K-AKT-mTOR pathway and prostate cancer: At the crossroads of AR, MAPK, and WNT signaling. Int J Mol Sci. 21:45072020. View Article : Google Scholar : PubMed/NCBI
48	Kumari N, Reabroi S and North BJ: Unraveling the molecular nexus between GPCRs, ERS, and EMT. Mediators Inflamm. 2021:66554172021. View Article : Google Scholar : PubMed/NCBI