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ARID2 suppression promotes tumor progression and upregulates cytokeratin 8, 18 and β-4 integrin expression in TP53-mutated tobacco-related oral cancer and has prognostic implications

Cancer Gene Therapy volume 29, pages 1908–1917 (2022)Cite this article

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Abstract

Mutations in ARID2 and TP53 genes are found to be implicated in the tobacco related tumorigeneses. However, the effect of loss of ARID2 in the TP53 mutated background in tobacco related cancer including oral cancer has not been investigated yet. Hence, in this study we knockdown ARID2 using shRNA mediated knockdown strategy in TP53 mutated oral squamous cell carcinoma (OSCC) cell line and studied its tumorigenic role. Our study revealed that suppression of ARID2 in TP53 mutated oral cancer cells increases cell motility and invasion, induces drastic morphological changes and leads to a marked increase in the expression levels of cytokeratins, and integrins, CK8, CK18 and β4-Integrin, markers of cell migration/invasion in oral cancer. ARID2 suppression also showed early onset and increased tumorigenicity in-vivo. Interestingly, transcriptome profiling revealed differentially expressed genes associated with migration and invasion in oral cancer cells including AKR1C2, NCAM2, NOS1, ADAM23 and genes of S100A family in ARID2 knockdown TP53 mutated oral cancer cells. Pathway analysis of differentially regulated genes identified “cancer pathways” and “PI3K/AKT Pathway” to be significantly dysregulated upon suppression of ARID2 in TP53 mutated OSCC cells. Notably, decreased ARID2 expression and increased CK8, CK18 expression leads to poor prognosis in Head and Neck cancer (HNSC) patients as revealed by Pan-Cancer TCGA data analysis. To conclude, our study is the first to demonstrate tumor suppressor role of ARID2 in TP53 mutated background indicating their cooperative role in OSCC, and also highlights its prognostic implications suggesting ARID2 as an important therapeutic target in OSCC.

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Fig. 1: shRNA mediated ARID2 knockdown in ITOC-02 cell line using pLKO vector.
Fig. 2: Suppression of ARID2 in TP53 mutated background leads to enhanced tumorigenic potential in-vivo in OSCC.
Fig. 3: ARID2 knockdown leads to change in morphology of the cells as examined by phase contrast microscopy and transmission electron microscopy.
Fig. 4: ARID2 knockdown in TP53 mutated OSCC upregulates markers of neoplastic progression.
Fig. 5: RNA sequencing analysis revealed altered expression of genes involved in migration and invasion in ARID2 knockdown TP53 mutated OSCC.
Fig. 6: Pan-Cancer TCGA analysis revealed poor prognosis associated with decreased expression of ARID2 andincreased expression of CK8, CK18, β4-Integrin in HNSC patients.

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Most data have been incorporated into the article and its online supplementary material. Remaining data are available on reasonable request to corresponding author.

References

  1. Shain AH, Pollack JR. The spectrum of SWI/SNF mutations, ubiquitous in human cancers. PLoS One. 2013;8:e55119.

    Article  Google Scholar 

  2. Cohet N, Stewart KM, Mudhasani R, Asirvatham AJ, Mallappa C, Imbalzano KM, et al. SWI/SNF chromatin remodeling enzyme ATPases promote cell proliferation in normal mammary epithelial cells. J Cell Physiol. 2010;223:667–78.

    CAS  Google Scholar 

  3. Imbalzano AN, Kwon H, Green MR, Kingston RE. Facilitated binding of TATA-binding protein to nucleosomal DNA. Nature 1994;370:481–5.

    Article  CAS  Google Scholar 

  4. Kwon H, Imbalzano AN, Khavari PA, Kingston RE, Green MR. Nucleosome disruption and enhancement of activator binding by a human SW1/SNF complex. Nature 1994;370:477–81.

    Article  CAS  Google Scholar 

  5. Yan Z, Cui K, Murray DM, Ling C, Xue Y, Gerstein A, et al. PBAF chromatin-remodeling complex requires a novel specificity subunit, BAF200, to regulate expression of selective interferon-responsive genes. Genes Dev. 2005;19:1662–7.

    Article  CAS  Google Scholar 

  6. Olivier M, Hollstein M, Hainaut P. TP53 mutations in human cancers: origins, consequences, and clinical use. Cold Spring Harb Perspect Biol. 2010;2:a001008.

    Article  Google Scholar 

  7. Loesch R, Chenane L, Colnot S. ARID2 chromatin remodeler in hepatocellular carcinoma. Cells 2020;9:2152.

    Article  CAS  Google Scholar 

  8. Manceau G, Letouzé E, Guichard C, Didelot A, Cazes A, Corté H, et al. Recurrent inactivating mutations of ARID2 in non-small cell lung carcinoma. Int J Cancer. 2013;132:2217–21.

    Article  CAS  Google Scholar 

  9. Maitra A, Biswas N, Amin K, Kowtal P, Kumar S, Das S, et al. Mutational landscape of gingivo-buccal oral squamous cell carcinoma reveals new recurrently-mutated genes and molecular subgroups. Nat Commun. 2013;4:2873.

    Article  Google Scholar 

  10. Fadlullah MZ, Chiang IK, Dionne KR, Yee PS, Gan CP, Sam KK, et al. Genetically-defined novel oral squamous cell carcinoma cell lines for the development of molecular therapies. Oncotarget 2016;7:27802–18.

    Article  Google Scholar 

  11. Pansare K, Gardi N, Kamat S, Dange P, Previn R, Gera P, et al. Establishment and genomic characterization of gingivobuccal carcinoma cell lines with smokeless tobacco associated genetic alterations and oncogenic PIK3CA mutation. Sci Rep. 2019;9:8272.

    Article  Google Scholar 

  12. Silwal-Pandit L, Vollan HK, Chin SF, Rueda OM, McKinney S, Osako T, et al. TP53 mutation spectrum in breast cancer is subtype specific and has distinct prognostic relevance. Clin Cancer Res. 2014;20:3569–80.

    Article  CAS  Google Scholar 

  13. Gong F, Fahy D, Liu H, Wang W, Smerdon MJ. Role of the mammalian SWI/SNF chromatin remodeling complex in the cellular response to UV damage. Cell Cycle. 2008;7:1067–74.

    Article  CAS  Google Scholar 

  14. Wu RC, Wang TL, Shih IM. The emerging roles of ARID1A in tumor suppression. Cancer Biol Ther. 2014;15:655–64.

    Article  Google Scholar 

  15. Cajuso T, Hänninen UA, Kondelin J, Gylfe AE, Tanskanen T, Katainen R, et al. Exome sequencing reveals frequent inactivating mutations in ARID1A, ARID1B, ARID2, and ARID4A in microsatellite unstable colorectal cancer. Int J Cancer. 2014;135:611–23.

    Article  CAS  Google Scholar 

  16. Kadoch C, Hargreaves DC, Hodges C, et al. Proteomic and bioinformatic analysis of mammalian swi/snf complexes identifies extensive roles in human malignancy. Nat Genet. 2013;45:592–601.

    Article  CAS  Google Scholar 

  17. Duan Y, Tian L, Gao Q, Liang L, Zhang W, Yang Y, et al. Chromatin remodeling gene ARID2 targets cyclin D1 and cyclin E1 to suppress hepatoma cell progression. Oncotarget 2016;7:45863–75.

    Article  Google Scholar 

  18. Jiang H, Cao HJ, Ma N, Bao WD, Wang JJ, Chen TW, et al. Chromatin remodeling factor ARID2 suppresses hepatocellular carcinoma metastasis via DNMT1-Snail axis. Proc Natl Acad Sci USA. 2020;117:4770–80.

    Article  CAS  Google Scholar 

  19. Zhang L, Wang W, Li X, He S, Jianfeng Y, Wang X, et al. MicroRNA-155 promotes tumor growth of human hepatocellular carcinoma by targeting ARID2. Int J Oncol. 2016;48:2425–34.

    Article  CAS  Google Scholar 

  20. Liu X, Zhang H, Zhou P, Yu Y, Zhang H, Chen L, et al. CREB1 acts via the miR-922/ARID2 axis to enhance malignant behaviour of liver cancer cells. Oncol Rep. 2021;45:79.

    Article  Google Scholar 

  21. Zhou C, Zhang Y, Yan R, Huang L, Mellor AL, Yang Y, et al. Exosome-derived miR-142-5p remodels lymphatic vessels and induces IDO to promote immune privilege in the tumour microenvironment. Cell Death Differ. 2021;28:715–29.

    Article  CAS  Google Scholar 

  22. Wu M, Duan Q, Liu X, Zhang P, Fu Y, Zhang Z, et al. MiR-155-5p promotes oral cancer progression by targeting chromatin remodeling gene ARID2. Biomed Pharmacother. 2020;122:109696.

    Article  CAS  Google Scholar 

  23. Moreno T, Monterde B, González-Silva L, Betancor-Fernández I, Revilla C, Agraz-Doblas A, et al. ARID2 deficiency promotes tumor progression and is associated with higher sensitivity to chemotherapy in lung cancer. Oncogene 2021;40:2923–35.

    Article  CAS  Google Scholar 

  24. Fukumoto T, Lin J, Fatkhutdinov N, Liu P, Somasundaram R, Herlyn M, et al. ARID2 deficiency correlates with the response to immune checkpoint blockade in melanoma. J Invest Dermatol. 2020;S0022-202X:32396–4.

    Google Scholar 

  25. Goh AY, Ramlogan-Steel CA, Jenkins KS, Steel JC, Layton CJ. Presence and prevalence of UV related genetic mutations in uveal melanoma: similarities with cutaneous melanoma. Neoplasma 2020;67:958–71.

    Article  CAS  Google Scholar 

  26. Williams EA, Shah N, Montesion M, Sharaf R, Pavlick DC, Sokol ES, et al. Melanomas with activating RAF1 fusions: clinical, histopathologic, and molecular profiles. Mod Pathol. 2020;33:1466–74.

    Article  CAS  Google Scholar 

  27. Saqcena M, Leandro-Garcia LJ, Maag JLV, Tchekmedyian V, Krishnamoorthy GP, Tamarapu. SWI/SNF complex mutations promote thyroid tumor progression and insensitivity to redifferentiation therapies. Cancer Disco. 2020;11:1158–75.

    Article  Google Scholar 

  28. Oba A, Shimada S, Akiyama Y, Nishikawaji T, Mogushi K, Ito H, et al. ARID2 modulates DNA damage response in human hepatocellular carcinoma cells. J Hepatol. 2017;66:942–51.

    Article  CAS  Google Scholar 

  29. Pan D, Kobayashi A, Jiang P, Ferrari de Andrade L, Tay RE, Luoma AM, et al. A major chromatin regulator determines resistance of tumor cells to T cell-mediated killing. Sci. 2018;359:770–5.

    Article  CAS  Google Scholar 

  30. Fuchs E, Cleveland DW. A structural scaffolding of intermediate filaments in health and disease. Sci. 1998;279:514–9.

    Article  CAS  Google Scholar 

  31. Paramio JM, Jorcano JL. Beyond structure: do intermediate filaments modulate cell signalling? Bioessays 2002;24:836–44.

    Article  CAS  Google Scholar 

  32. Moll R, Divo M, Langbein L. The human keratins: biology and pathology. Histochem Cell Biol. 2008;129:705–33.

    Article  CAS  Google Scholar 

  33. Vaidya MM, Borges AM, Pradhan SA, Bhisey AN. Cytokeratin expression in squamous cell carcinomas of the tongue and alveolar mucosa. Eur J Cancer B Oral Oncol. 1996;32B:333–6.

    Article  CAS  Google Scholar 

  34. Borradori L, Sonnenberg A. Hemidesmosomes: roles in adhesion, signalling and human diseases. Curr Opin Cell Biol 1996;8:647–56.

    Article  CAS  Google Scholar 

  35. Yang Y, Dowling J, Yu QC, Kouklis P, Cleveland DW, Fuchs E. An essential cytoskeletal linker protein connecting actin microfilaments to intermediate filaments. Cell 1996;86:655–65.

    Article  CAS  Google Scholar 

  36. Giancotti FG. Targeting integrin beta4 for cancer and anti-angiogenic therapy. Trends Pharmacol Sci 2007;28:506–11.

    Article  CAS  Google Scholar 

  37. Alam H, Kundu ST, Dalal SN, Vaidya MM. Loss of keratins 8 and 18 leads to alterations in α6β4-integrin-mediated signalling and decreased neoplastic progression in an oral-tumour-derived cell line. J Cell Sci. 2011;124:2096–106.

    Article  CAS  Google Scholar 

  38. Kippenberger S, Hofmann M, Zöller N, Thaçi D, Müller J, Kaufmann R, et al. A. Ligation of beta4 integrins activates PKB/Akt and ERK1/2 by distinct pathways-relevance of the keratin filament. Biochem Biophys Acta. 2010;1803:940–50.

    Article  CAS  Google Scholar 

  39. Simbolo M, Vicentini C, Ruzzenente A, Brunelli M, Conci S, Fassan M, et al. Genetic alterations analysis in prognostic stratified groups identified TP53 and ARID1A as poor clinical performance markers in intrahepatic cholangiocarcinoma. Sci Rep. 2018;8:7119.

    Article  Google Scholar 

  40. Zhang J, Hou S, You Z, Li G, Xu S, Li X, et al. Expression and prognostic values of ARID family members in breast cancer. Aging (Albany NY). 2021;13:5621–37.

    Article  CAS  Google Scholar 

  41. Afrem M, CrăiŢoiu Ş, Hîncu M, Manolea H, Nicolae V, CrăiŢoiu M. Study of CK18 and GDF5 immunoexpression in oral squamous cell carcinoma and their prognostic value. Rom J Morphol Embryol. 2016;57:167–72.

    Google Scholar 

  42. Nagata M, Noman A, Suzuki K, Kurita H, Ohnishi M, Ohyama T, et al. ITGA3 and ITGB4 expression biomarkers estimate the risks of locoregional and hematogenous dissemination of oral squamous cell carcinoma. BMC Cancer. 2013;13:410.

    Article  Google Scholar 

  43. Choudhari SK, Chaudhary M, Bagde S, Gadbail AR, Joshi V. Nitric oxide and cancer: a review. World J Surg Oncol. 2013;11:118.

    Article  Google Scholar 

  44. Raab JR, Resnick S, Magnuson T. Genome-wide transcriptional regulation mediated by biochemically distinct SWI/SNF complexes. PLoS Genet. 2015;11:e1005748.

    Article  Google Scholar 

  45. Pfister NT, Fomin V, Regunath K, Zhou JY, Zhou W, Silwal-Pandit L, et al. Mutant p53 cooperates with the SWI/SNF chromatin remodeling complex to regulate VEGFR2 in breast cancer cells. Genes Dev. 2015;29:1298–315.

    Article  CAS  Google Scholar 

  46. Song H, Hollstein M, Xu Y. p53 gain-of-function cancer mutants induce genetic instability by inactivating ATM. Nat Cell Biol. 2007;9:573–80.

    Article  CAS  Google Scholar 

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Acknowledgements

We would like to thank Genomics & NGS facility, electron microscopy facility, animal House, biorepository, ACTREC, Dr. Pradnya Kowtal and members of Sarin Lab. We are also grateful to TCGA/ULACAN databases to make data available free for analysis. We also thank NIBMG, and grant support from Department of Biotechnology, Govt. of India (Grant Number (BT/MB/01/ICGC/2009). We are grateful to Dr. T. Mahmoudi, Netherland for providing ARID2 antibody and Dr. Sorab Dalal, ACTREC, Mumbai for providing pLKO-PURO EGFP vector. The study has been supported by grant from Department of Biotechnology, Govt. of India (Grant Number (BT/MB/01/ICGC/2009) under the International Cancer Genome Consortium (ICGC) project.

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  1. These authors contributed equally: Pallavi Shukla, Prerana Dange.

Authors and Affiliations

  1. Sarin Lab, Advanced Centre for Treatment Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, India

    Pallavi Shukla, Prerana Dange & Rajiv Sarin

  2. Comparative Oncology & Small Animal Imaging Facility, Advanced Centre for Treatment Research and Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, India

    Bhabani Shankar Mohanty & Pradip Chaudhari

  3. Bioinformatics Centre, Advanced Centre for Treatment, Research & Education in Cancer (ACTREC), Tata Memorial Centre, Kharghar, Navi Mumbai, India

    Nikhil Gadewal

  4. Homi Bhabha National Institute, Training School Complex, Anushakti Nagar, Mumbai, India

    Pradip Chaudhari & Rajiv Sarin

  5. Cancer Genetics Clinic, Tata Memorial Hospital, Tata Memorial Centre, Mumbai, India

    Rajiv Sarin

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Contributions

PS conceptualized the idea, conducted cloning and molecular biology experiments, TCGA data analysis, designed and wrote the manuscript. PD performed cell culture and all functional assays and helped in animal experiments. PC supervised BSM in conducting all animal experiments. NG analyzed bioinformatics data. RS conceived the project, supervised all the experiments and edited the manuscript.

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Correspondence to Rajiv Sarin.

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Shukla, P., Dange, P., Mohanty, B.S. et al. ARID2 suppression promotes tumor progression and upregulates cytokeratin 8, 18 and β-4 integrin expression in TP53-mutated tobacco-related oral cancer and has prognostic implications. Cancer Gene Ther 29, 1908–1917 (2022). https://doi.org/10.1038/s41417-022-00505-x

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