Modulation of PD-L1 Expression by Cisplatin and Gemcitabine in Bladder Cancer Cells

Öz

Bladder cancer (BC) incidence is rising globally, despite available treatment options, including tumor resection, systemic chemotherapy, and immune checkpoint inhibitors targeting PD-1/PD-L1. Gemcitabine and cisplatin are commonly combined as first-line systemic therapy, yet their effects on PD-L1 expression remain unclear. This study evaluated the effects of cisplatin and gemcitabine, alone and combined, on PD-L1 expression in a BC cell line, RT-4, and a normal epithelial cell line, ARPE-19. Cells were treated with IC20 doses of each drug, and PD-L1 expression was quantified by qRT-PCR after 48 hours. In ARPE-19 cells, cisplatin treatment induced a dose-dependent PD-L1 increase, peaking at a 59.6-fold elevation at higher doses (p < 0.001), while gemcitabine alone showed no effect. The combination of low-dose cisplatin and gemcitabine further elevated PD-L1 expression (25.2-fold, p < 0.05). In contrast, in RT-4 cells, low-dose cisplatin downregulated PD-L1, but higher doses resulted in a 61.3-fold increase (p < 0.05). Gemcitabine alone downregulated PD-L1 in RT-4 cells; however, the combined treatment produced a 99.4-fold upregulation (p < 0.001). These findings highlight dose- and cell type-specific effects of cisplatin and gemcitabine on PD-L1 regulation, with combination therapy notably upregulating PD-L1 in BC cells. This underscores the potential for integrating chemotherapy with immune checkpoint inhibitors to counteract chemotherapy-induced immune evasion in BC. Differential responses between normal and cancer cells emphasize the need for tumor-specific therapeutic approaches and further research on PD-L1 regulation to optimize immune checkpoint therapy.

Anahtar Kelimeler

• Bladder cancer
• PD-L1
• cisplatin
• gemcitabine
• combination therapy

Modulation of PD-L1 Expression by Cisplatin and Gemcitabine in Bladder Cancer Cells

Öz

Bladder cancer (BC) incidence is rising globally, despite available treatment options, including tumor resection, systemic chemotherapy, and immune checkpoint inhibitors targeting PD-1/PD-L1. Gemcitabine and cisplatin are commonly combined as first-line systemic therapy, yet their effects on PD-L1 expression remain unclear. This study evaluated the effects of cisplatin and gemcitabine, alone and combined, on PD-L1 expression in a BC cell line, RT-4 and normal epithelial cell line, ARPE-19. Cells were treated with IC20 doses of each drug, and PD-L1 expression was quantified by qRT-PCR after 48 hours. In ARPE-19 cells, cisplatin treatment induced a dose-dependent PD-L1 increase, peaking at a 59.6-fold elevation at higher doses (p < 0.001), while gemcitabine alone showed no effect. The combination of low-dose cisplatin and gemcitabine further elevated PD-L1 expression (25.2-fold, p < 0.05). In contrast, in RT-4 cells, low-dose cisplatin downregulated PD-L1, but higher doses resulted in a 61.3-fold increase (p < 0.05). Gemcitabine alone downregulated PD-L1 in RT-4 cells; however, the combined treatment produced a 99.4-fold upregulation (p < 0.001). These findings highlight dose- and cell type-specific effects of cisplatin and gemcitabine on PD-L1 regulation, with combination therapy notably upregulating PD-L1 in BC cells. This underscores the potential for integrating chemotherapy with immune checkpoint inhibitors to counteract chemotherapy-induced immune evasion in BC. Differential responses between normal and cancer cells emphasize the need for tumor-specific therapeutic approaches and further research on PD-L1 regulation to optimize immune checkpoint therapy.

Anahtar Kelimeler

• Bladder cancer
• PD-L1
• cisplatin
• gemcitabine
• combination therapy

Kaynakça

1. [1.] Bray F, Ferlay J, et al. 2018. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians; 68: 394-424.
2. [2.] Martínez-Lacaci I, García Morales P, et al. 2007. Tumour cells resistance in cancer therapy. Clinical Translational Oncology; 9: 13-20.
3. [3.] Kaufman DS, Shipley WU, et al. 2009. Bladder cancer. The Lancet; 374: 239-249.
4. [4.] Sternberg CN, Bellmunt J, et al. 2013. ICUD-EAU international consultation on bladder cancer 2012: chemotherapy for urothelial carcinoma—neoadjuvant and adjuvant settings. European Urology; 63: 58-66.
5. [5.] Patel SP, Kurzrock R. 2015. PD-L1 expression as a predictive biomarker in cancer immunotherapy. Molecular Cancer Therapeutics; 14: 847-856.
6. [6.] Doroshow DB, Bhalla S, et al. 2021. PD-L1 as a biomarker of response to immune-checkpoint inhibitors. Nature Reviews Clinical Oncology; 18: 345-362.
7. [7.] Herbst RS, Soria JC, et al. 2014. Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A in cancer patients. Nature; 515: 563-567.
8. [8.] Galsky MD, Daneshmand S, et al. 2023. Gemcitabine and cisplatin plus nivolumab as organ-sparing treatment for muscle-invasive bladder cancer: a phase 2 trial. Nature Medicine; 29: 2825-2834.

9. [9.] Kelley RK, Ueno M, et al. 2023. Pembrolizumab in combination with gemcitabine and cisplatin compared with gemcitabine and cisplatin alone for patients with advanced biliary tract cancer (KEYNOTE-966): a randomised, double-blind, placebo-controlled, phase 3 trial. The Lancet; 401: 1853-1865.
10. [10.] Sevim Nalkiran H, Akcora Yildiz D, et al. 2023. Targeting the anaphase-promoting complex/cyclosome (APC/C) enhanced antiproliferative and apoptotic response in bladder cancer. Saudi Journal of Biological Sciences; 30: 103564.
11. [11.] Durur S, Uzun H, et al. 2023. Prognostic importance of PD-L1 expression in bladder cancer patients. Journal of Cancer Research and Therapeutics; 19: S633-S638.
12. [12.] Chen M, Wang S. 2024. Preclinical development and clinical studies of targeted JAK/STAT combined Anti-PD-1/PD-L1 therapy. International Immunopharmacology; 130: 111717.
13. [13.] Antonangeli F, Natalini A, et al. 2020. Regulation of PD-L1 Expression by NF-κB in Cancer. Frontiers in Immunology; 11: 584626.
14. [14.] Kakoti S, Sato H, et al. 2020. DNA Repair and Signaling in Immune-Related Cancer Therapy. Frontiers in Molecular Biosciences; 7.
15. [15.] Babjuk M, Böhle A, et al. 2017. EAU guidelines on non–muscle-invasive urothelial carcinoma of the bladder: update 2016. European Urology; 71: 447-461.
16. [16.] Bellmunt J, De Wit R, et al. 2017. Pembrolizumab as second-line therapy for advanced urothelial carcinoma. New England Journal of Medicine; 376: 1015-1026.
17. [17.] Powles T, Park SH, et al. 2020. Avelumab maintenance therapy for advanced or metastatic urothelial carcinoma. New England Journal of Medicine; 383: 1218-1230.
18. [18.] Yoshino H, Yokoyama S, et al. 2023. Characterization and treatment of gemcitabine- and cisplatin-resistant bladder cancer cells with a pan-RAS inhibitor. FEBS Open Bio; 13: 1056-1066.
19. [19.] Aldossary S. 2019. Review on Pharmacology of Cisplatin: Clinical Use, Toxicity and Mechanism of Resistance of Cisplatin. Biomedical and Pharmacology Journal; 11: 07-15.
20. [20.] de Biasi AR, Villena-Vargas J, et al. 2014. Cisplatin-induced antitumor immunomodulation: a review of preclinical and clinical evidence. Clinical Cancer Research: An Official Journal of the American Association for Cancer; 20: 5384-5391.
21. [21.] Qin X, Liu C, et al. 2010. Cisplatin induces programmed death-1-ligand 1 (PD-L1) over-expression in hepatoma H22 cells via Erk/MAPK signaling pathway. Cellular Molecular Biology; 56: 1366-1372.
22. [22.] Zhang P, Su D-M, et al. 2008. Chemopreventive agents induce programmed death-1-ligand 1 (PD-L1) surface expression in breast cancer cells and promote PD-L1-mediated T cell apoptosis. Molecular Immunology; 45: 1470-1476.
23. [23.] Crinò L, Scagliotti G, et al. 1997. Cisplatin-gemcitabine combination in advanced non-small-cell lung cancer: a phase II study. Journal of Clinical Oncology: Official Journal of the American Society of Clinical Oncology; 15: 297-303.
24. [24.] von der Maase H, Sengelov L, et al. 2005. Long-term survival results of a randomized trial comparing gemcitabine plus cisplatin, with methotrexate, vinblastine, doxorubicin, plus cisplatin in patients with bladder cancer. Journal of Clinical Oncology: Official Journal of the American Society of Clinical Oncology; 23: 4602-4608.
25. [25.] Hitt R, Castellano D, et al. 1998. Phase II trial of cisplatin and gemcitabine in advanced squamous-cell carcinoma of the head and neck. Annals of Oncology: Official Journal of the European Society for Medical Oncology; 9: 1347-1349.
26. [26.] Nagourney RA, Flam M, et al. 2008. Carboplatin plus Gemcitabine Repeating Doublet Therapy in Recurrent Breast Cancer. Clinical Breast Cancer; 8: 432-435.
27. [27.] Pfisterer J, Plante M, et al. 2006. Gemcitabine plus carboplatin compared with carboplatin in patients with platinum-sensitive recurrent ovarian cancer: an intergroup trial of the AGO-OVAR, the NCIC CTG, and the EORTC GCG. Journal of Clinical Oncology; 24: 4699-4707.
28. [28.] Reck M, von Pawel J, et al. 2009. Phase III trial of cisplatin plus gemcitabine with either placebo or bevacizumab as first-line therapy for nonsquamous non–small-cell lung cancer: AVAiL. Journal of Clinical Oncology; 27: 1227-1234.
29. [29.] Emens LA, Middleton G. 2015. The interplay of immunotherapy and chemotherapy: harnessing potential synergies. Cancer immunology research; 3: 436-443.
30. [30.] Hanna GG, Coyle VM, et al. 2015. Immune modulation in advanced radiotherapies: targeting out-of-field effects. Cancer Letters; 368: 246-251.
31. [31.] Dong H, Strome SE, et al. 2002. Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion. Nature Medicine; 8: 793-800.
32. [32.] Iwai Y, Ishida M, et al. 2002. Involvement of PD-L1 on tumor cells in the escape from host immune system and tumor immunotherapy by PD-L1 blockade. Proceedings of the National Academy of Sciences; 99: 12293-12297.
33. [33.] Thompson RH, Kuntz SM, et al. 2006. Tumor B7-H1 is associated with poor prognosis in renal cell carcinoma patients with long-term follow-up. Cancer Research; 66: 3381-3385.
34. [34.] Wu C, Zhu Y, et al. 2006. Immunohistochemical localization of programmed death-1 ligand-1 (PD-L1) in gastric carcinoma and its clinical significance. Acta Histochemica; 108: 19-24.
35. [35.] Ghebeh H, Mohammed S, et al. 2006. The B7-H1 (PD-L1) T lymphocyte-inhibitory molecule is expressed in breast cancer patients with infiltrating ductal carcinoma: correlation with important high-risk prognostic factors. Neoplasia; 8: 190-198.
36. [36.] Fu D, Wu J, et al. 2020. T cell recruitment triggered by optimal dose platinum compounds contributes to the therapeutic efficacy of sequential PD-1 blockade in a mouse model of colon cancer. American Journal of Cancer Research; 10: 473.
37. [37.] Hu J, Kinn J, et al. 2013. The effects of chemotherapeutic drugs on human monocyte-derived dendritic cell differentiation and antigen presentation. Clinical Experimental Immunology; 172: 490-499.
38. [38.] Tran L, Allen CT, et al. 2017. Cisplatin alters antitumor immunity and synergizes with PD-1/PD-L1 inhibition in head and neck squamous cell carcinoma. Cancer Immunology Research; 5: 1141-1151.
39. [39.] Sawasdee N, Thepmalee C, et al. 2020. Gemcitabine enhances cytotoxic activity of effector T-lymphocytes against chemo-resistant cholangiocarcinoma cells. International Immunopharmacology; 78: 106006.
40. [40.] Oing C, Rink M, et al. 2016. Second line chemotherapy for advanced and metastatic urothelial carcinoma: vinflunine and beyond—a comprehensive review of the current literature. The Journal of Urology; 195: 254-263.