Changes in expression of TLR-4, TGF-β, INF-γ, TNF-α in cultured T24/83 cells of invasive bladder cancer treated with cisplatin and/or polyphenolic adjuvant melanin
DOI:
https://doi.org/10.32471/exp-oncology.2312-8852.vol-43-no-1.15739Keywords:
apoptosis, cell line T24/83, INF-γ, invasive bladder cancer, melanin, TGFβ, TLR4, TNF, urothelial carcinomaAbstract
Background: Toll-like receptor 4 (TLR4) is known to be involved in carcinogenesis and cancer progression. Changes in TLR4 expression are associated with changes in the expression of key cellular cytokines (transforming growth factor-β (TGF-β), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ)), which affect cancer progression and metastasis. Aim: To study changes in the expression of TLR4, TGF-β, TNF-α, IFN-γ genes, the level of apoptosis and cell cycle distribution in human invasive urothelial carcinoma T24/83 cells under the treatment with polyphenolic adjuvant compound of fungal origin melanin, cytotoxic drug cisplatin, and combination of both. Materials and Methods: T24/83 cells were incubated with cisplatin (0.05 mM), melanin (5 µg/ml), or their combination. The expression level of TLR-4, TGF-β, INF-γ, TNF-α was evaluated by the real time polymerase chain reaction. The flow cytometry was used to study cell cycle distribution, proliferative activity and level of apoptosis. Morphological analysis of the Т24/83 cells was performed as well. Results: Melanin, cisplatin, and their combination downregulate TLR4 expression (2.67; 1.28; and 2.73-fold decrease, respectively) and TNF-α expression (6.5; 1.4; and 1.7-fold decrease, respectively). Melanin did not affect TGF-β expression while cisplatin caused 13-fold downregulation of TGF-β. The combined use of cisplatin and melanin decreased TGF-β expression by 6.5 times. The upregulation of IFN-γ by melanin, cisplatin, and their combination was demonstrated (4.3; 6.7; and 2-fold increase, respectively). All treatment modalities increased the level of apoptosis in T24/83 cells. Melanin treatment increased significantly the proportion of fibroblast-like cells in T24/83 culture with decreased cell adhesion to the substrate. Conclusions: Melanin, cisplatin, and combination of both agents affect significantly TLR4, TNF-α, TGF-β, INF-γ expression, cell cycle distribution and morphology in T24/83 cells suggesting their transition to less aggressive phenotype.
References
Shcheblyakov DV, Logunov DY, Tukhvatulin AI, et al. Toll-Like receptors (TLRs): The role in tumor progression. Acta Naturae 2010; 2: 21–9.
Kawai T, Akira S. Toll-like receptors and their crosstalk with other innate receptors in infection and immunity. Immunity 2011; 34: 637–50.
Wolska A, Lech-Marańda E, Robak T. Toll-like receptors and their role in carcinogenesis and anti-tumor treatment. Cell Mol Biol Lett 2009; 14: 248–72.
LaRue H, Ayari C, Bergeron A, et al. Toll-like receptors in urothelial cells -targets for cancer immunotherapy. Nat Rev Urol 2013; 10: 537–45.
Huang B, Zhao J, Li H, et al. Toll-like receptors on tumor cells facilitate evasion of immune surveillance. Cancer Res 2005; 65: 5009–14.
Salaun B, Romero P, Lebecque S. Toll-like receptors’ two-edged sword: when immunity meets apoptosis. Eur J Immunol 2007; 37: 3311–8.
Takeda K, Kaisho T, Akira S. Toll-like receptors. Annu Rev Immunol 2003; 21: 335–76.
Ohadian Moghadam S, Nowroozi MR. Toll-like receptors: The role in bladder cancer development, progression and immunotherapy. Scand J Immunol 2019; 90: e12818.
Oblak A, Jerala R. Toll-like receptor 4 activation in cancer progression and therapy. Clin Dev Immunol 2011. Article ID: 609579. doi: 10.1155/2011/609579.
Awasthi S. Toll-like receptor-4 modulation for cancer immunotherapy. Front Immunol 2014; 5: 328.
Pradere JP, Dapito DH, Schwabe RF. The Yin and Yang of Toll-like receptors in cancer. Oncogene 2014; 33: 3485–95.
Fukata M, Shang L, Santaolalla R, et al. Constitutive activation of epithelial TLR4 augments inflammatory responses to mucosal injury and drives colitis-associated tumorigenesis. Inflamm Bowel Dis 2011; 17: 1464–73.
Wu Q, Wu W, Fu B, et al. JNK signaling in cancer cell survival. Med Res Rev 2019; 39: 2082–104.
Tang X, Zhu Y. TLR4 signaling promotes immune escape of human colon cancer cells by inducing immunosuppressive cytokines and apoptosis resistance. Oncol Res 2012; 20: 15–24.
Lu YC, Yeh WC, Ohashi PS. LPS/TLR4 signal transduction pathway. Cytokine 2008; 42: 145–51.
Kelsh RM, McKeown-Longo PJ. Topographical changes in extracellular matrix: Activation of TLR4 signaling and solid tumor progression. Trends Cancer Res 2013; 9: 1–13.
Ikebe M, Kitaura Y, Nakamura M, et al. Lipopolysaccharide (LPS) increases the invasive ability of pancreatic cancer cells through the TLR4/MyD88 signaling pathway. J Surg Oncol 2009; 100: 725–31.
Earl TM, Nicoud IB, Pierce JM, et al. Silencing of TLR4 decreases liver tumor burden in a murine model of colorectal metastasis and hepatic steatosis. Ann Surg Oncol 2009; 16: 1043–50.
Li C, Li H, Jiang K, et al. TLR4 signaling pathway in mouse Lewis lung cancer cells promotes the expression of TGF-β1 and IL-10 and tumor cells migration. Biomed Mater Eng 2014; 24: 869–75.
Ni L, Lu J. Interferon gamma in cancer immunotherapy. Cancer Med 2018; 7: 4509–16.
Al-Obeed O, El-Obeid AS, Matou-Nasri S, et al. Herbal melanin inhibits colorectal cancer cell proliferation by altering redox balance, inducing apoptosis, and modulating MAPK signaling. Cancer Cell Int 2020; 20: 126.
Kondratiuk T, Akulenko T, Beregova T, et al. Microorganisms, perspective for biotechnology, medicine, environmenal technologies, in the collection of microscopic fungi ESC «Institute of biology and medicine», Taras Shevchenko national university of Kyiv. Bull Taras Shevchenko Nat Univ Kyiv Series Biol 2017; 73: 31–36.
Nicoletti I, Migliorati G, Pagliacci MC, et al. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 1991; 139: 271–9.
Sarnatskaya V, Shlapa Y, Yushko L, et al. Biological activity of cerium dioxide nanoparticles. J Biomed Mater Res A 2020; 108: 1703–12.
Sato Y, Goto Y, Narita N, et al. Cancer cells expressing Toll-like receptors and the tumor microenvironment. Cancer Microenviron 2009; 2: 205–14.
Yusuf N, Nasti TH, Long JA, et al. Protective role of Toll-like receptor 4 during the initiation stage of cutaneous chemical carcinogenesis. Cancer Res 2008; 68: 615–22.
Rakoff-Nahoum S, Medzhitov R. Toll-like receptors and cancer. Nat Rev Cancer 2009; 9: 57–63.
Kato T, Ishikawa K, Nemoto R, et al. Morphological characterization of two established cell lines, T24 and MGH-U1, derived from human urinary bladder carcinoma. Tohoku J Exp Med 1978; 124: 339–49.
Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell 2011; 144: 646–74.
Efstathiou JA, Mouw KW, Gibb EA, et al. Impact of immune and stromal infiltration on outcomes following bladder-sparing trimodality therapy for muscle-invasive bladder cancer. Eur Urol 2019; 76: 59–68.
Domenis R, Cifù A, Marinò D, et al. Toll-like Receptor-4 activation boosts the immunosuppressive properties of tumor cells-derived exosomes. Sci Rep 2019; 9: 8457.
Yakovlev P, Garmanchuk L, Falalyeyeva T, et al. Influence of the melanin from yeast-like fungi Nadsoniella nigra spр. X1 (i. Galindez, Antarctica) on proliferation, adhesion and apoptosis of tumor cells with epithelial origin. Likarsʹka sprava 2017; 5–6: 68–76.
Dasari S, Tchounwou PB. Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol 2014, 740: 364–78.
Yakovlev P, Garmanchuk L, Falalyeyeva T, et al. Cytostatic effect of polyphenol compound melanin in combination with Cisplatin on bladder cancer cell line T24/83.Eur Urol Suppl 2018; 17: e2553–4.
Yakovlev P, Klyushin D. Lymphocyte count in peripheral blood is a sensitive tool in pretreatment assessment of patients with urological cancer. Exp Oncol 2018; 40: 119–23.
Afsharimoghaddam A, Soleimani M, Lashay A, et al. Controversial roles played by toll like receptor 4 in urinary bladder cancer; A systematic review. Life Sci 2016; 158: 31–6.
Cai J, Yang MY, Hou N, et al. Association of tumor necrosis factor-α 308G/A polymorphism with urogenital cancer risk: a systematic review and meta-analysis. Genet Mol Res 2015; 14: 16102–12.
Josephs SF, Ichim TE, Prince SM, et al. Unleashing endogenous TNF-alpha as a cancer immunotherapeutic. J Transl Med 2018; 16: 242.
Sternberg CN, Arena MG, Pansadoro V, et al. Recombinant tumor necrosis factor for superficial bladder tumors. Ann Oncol 1992; 3: 741–5.
Wu NC, Wang JJ. Ibudilast, a phosphodiesterase inhibitor and Toll-like receptor-4 antagonist, improves hemorrhagic shock and reperfusion-Induced left ventricular dysfunction by reducing myocardial tumor necrosis factor α. Transplant Proc 2020; 52: 1869–74.
Singh A, Settleman J. EMT, cancer stem cells and drug resistance: an emerging axis of evil in the war on cancer. Oncogene 2010; 29: 4741–51.
Miyamoto H, Kubota Y, Shuin T, et al. Expression of transforming growth factor-beta 1 in human bladder cancer. Cancer 1995; 75: 2565–70.
Jakowlew SB. Transforming growth factor-beta in cancer and metastasis. Cancer Metastasis Rev2006; 25: 435–57.
Gupta S, Hau AM, Al-Ahmadie HA, et al. Transforming growth factor-β is an upstream regulator of mammalian target of rapamycin complex 2-dependent bladder cancer cell migration and invasion. Am J Pathol 2016; 186: 1351–60.
Kastan MB, Bartek J. Cell-cycle checkpoints and cancer. Nature 2004; 432: 316–23.
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