The activation of adenosine monophosphate–activated protein kinase inhibits the migration of tongue squamous cell carcinoma cells by targeting Claudin-1 via epithelial–mesenchymal transition

Abstract

Background: The role of Claudin-1 in tongue squamous cell carcinoma (TSCC) metastasis needs further clarification, particularly its impact on cell migration. Herein, our study aims to investigate the role of Claudin-1 in TSCC cell migration and its underlying mechanisms.

Methods: 36 TSCC tissue samples underwent immunohistochemical staining for Claudin-1. Western blotting and immunofluorescence analyses were conducted to evaluate Claudin-1 expression and distribution in TSCC cells. Claudin-1 knockdown cell lines were established using short hairpin RNA transfection. Migration effects were assessed through wound healing assays. Furthermore, the expression of EMT-associated molecules was measured via western blotting.

Results: Claudin-1 expression decreased as TSCC malignancy increased. Adenosine monophosphate–activated protein kinase (AMPK) activation led to increased Claudin-1 expression and membrane translocation, inhibiting TSCC cell migration and epithelial–mesenchymal transition (EMT). Conversely, Claudin-1 knockdown reversed these inhibitory effects on migration and EMT caused by AMPK activation.

Conclusions: Our results indicated that AMPK activation suppresses TSCC cell migration by targeting Claudin-1 and EMT pathways.

Keywords

AMPK / Claudin-1 / EMT / migration / tongue squamous cell carcinoma

References

[1] Rivera C. Essentials of oral cancer. Int J Clin Exp Pathol. 2015;8:11884.

[2] Hsu WH, Hua WJ, Qiu WL, Tseng AJ, Cheng HC, Lin TY. WSG, a glucose-enriche. polysaccharide from Ganoderma lucidum, suppresses tongue cancer cells via inhibition of EGFR-mediated signaling and potentiates cisplatin-induced apoptosis. Int J Biol Macromol. 2021;193(Pt B):1201-1208.

[3] Chan DA, Giaccia AJ. Hypoxia, gene expression, and metastasis. Cancer Metastasis Rev. 2007;26(2):333-339.

[4] Steeg PS. Targeting metastasis. Nat Rev Cancer. 2016;16(4):201-218.

[5] Schulzke JD, Günzel D, John LJ, Fromm M. Perspectives on tight junction research. Ann N Y Acad Sci. 2012;1257:1-19.

[6] Martin TA. The role of tight junctions in cancer metastasis. Semin Cell Dev Biol. 2014;36:224-231.

[7] Sawada N, Murata M, Kikuchi K, et al. Tight junctions and human diseases. Med Electron Microsc. 2003;36(3):147-156.

[8] Kolchakova D, Moten D, Batsalova T, Dzhambazov B. Tight junction protein Claudin-12 is involved in cell migration during metastasis. Biomolecules. 2021;11(5):636.

[9] Tsukita S, Yamazaki Y, Katsuno T, Tamura A, Tsukita S. Tight junction-based epithelial microenvironment and cell proliferation. Oncogene. 2008;27(55):6930-6938.

[10] Bhat AA, Ahmad R, Uppada SB, et al. Claudin-1 promotes TNF-α-induced epithelial-mesenchymal transition and migration in colorectal adenocarcinoma cells. Exp Cell Res. 2016;349(1):119-127.

[11] Oku N, Sasabe E, Ueta E, Yamamoto T, Osaki T. Tight junction protein claudin-1 enhances the invasive activity of oral squamous cell carcinoma cells by promoting cleavage of laminin-5 gamma2 chain via matrix metalloproteinase (MMP)-2 and membrane-type MMP-1. Cancer Res. 2006;66(10):5251-5257.

[12] Ouban A, Ahmed A. Analysis of the distribution and expression of claudin-1 tight junction protein in the oral cavity. Appl Immunohistochem Mol Morphol. 2015;23(6):444-448.

[13] Shen Z, Song W, Qian L, et al. Effect of claudin 1 on cell proliferation, migration and apoptosis in human cervical squamous cell carcinoma. Oncol Rep. 2021;45(2):606-618.

[14] Yamamoto D, Kayamori K, Sakamoto K, et al. Intracellular Claudin-1 at the invasive front of tongue squamous cell carcinoma is associated with lymph node metastasis. Cancer Sci. 2020;111(2):700-712.

[15] Sobel G, Paska C, Szabo I, et al. Increased expression of claudins in cervical squamous intraepithelial neoplasia and invasive carcinoma. Hum Pathol. 2005;36(2):162-169.

[16] Pastushenko I, Blanpain C. EMT transition states during tumor progression and metastasis. Trends Cell Biol. 2019;29(3):212-226.

[17] Hsu CC, Peng D, Cai Z, Lin HK. AMPK signaling and its targeting in cancer progression and treatment. Semin Cancer Biol. 2022;85:52-68.

[18] Rowart P, Wu J, Caplan MJ, Jouret F. Implications of AMPK in the formation of epithelial tight junctions. Int J Mol Sci. 2018;19(7):2040.

[19] Zhang L, Li J, Young LH, Caplan MJ. AMP-activated protein kinase regulates the assembly of epithelial tight junctions. Proc Natl Acad Sci USA. 2006;103(46):17272-17277.

[20] Guo XH, Lai XJ, Cai XL, et al. AICAR-induced activation of AMPK inhibits the migration of TSCC cells by targeting ZO-1. Oral Dis. 2020;26(1):228-233.

[21] Que T, Ren B, Fan Y, et al. Capsaicin inhibits the migration, invasion and EMT of renal cancer cells by inducing AMPK/mTOR-mediated autophagy. Chem Biol Interact. 2022;366:110043.

[22] Sun L, Cao J, Chen K, et al. Betulinic acid inhibits stemness and EMT of pancreatic cancer cells via activation of AMPK signaling. Int J Oncol. 2019;54(1):98-110.

[23] Bhat AA, Syed N, Therachiyil L, et al. Claudin-1, a double-edged sword in cancer. Int J Mol Sci. 2020;21(2):569.

[24] Chang JW, Seo ST, Im MA, et al. Claudin-1 mediates progression by regulating EMT through AMPK/TGF-β signaling in head and neck squamous cell carcinoma. Transl Res. 2022;247:58-78.

[25] Dos Reis PP, Bharadwaj RR, Machado J, et al. Claudin 1 overexpression increases invasion and is associated with aggressive histological features in oral squamous cell carcinoma. Cancer. 2008;113(11):3169-3180.

[26] Lourenco SV, Coutinho-Camillo CM, Buim ME, et al. Oral squamous cell carcinoma: status of tight junction claudins in the different histopathological patterns and relationship with clinical parameters. A tissue-microarray-based study of 136 cases. J Clin Pathol. 2010;63(7):609-614.

[27] Zheng B, Cantley LC. Regulation of epithelial tight junction assembly and disassembly by AMP-activated protein kinase. Proc Natl Acad Sci USA. 2007;104(3):819-822.

[28] Hsieh MJ, Chen JC, Yang WE, et al. Dehydroandrographolide inhibits oral cancer cell migration and invasion through NF-kappaB-, AP-1-, and SP-1-modulated matrix metalloproteinase-2 inhibition. Biochem Pharmacol. 2017;2017(130):10-20.

[29] Hashimoto I, Oshima T. Claudins and gastric cancer: an overview. Cancers (Basel). 2022;14(2):290.

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2024 The Authors. Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.