Skip to main content

Advertisement

SpringerLink
Log in

Thymoquinone enhanced the antitumor activity of cisplatin in human bladder cancer 5637 cells in vitro

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Purpose

Cisplatin-based chemotherapy is a primary alternative for treating bladder cancer. But drug resistance and various side effects are the main unsightliness challenges. In search of a novel chemotherapeutic approach, this study was conducted to investigate whether thymoquinone (TQ) chemosensitize 5637 bladder cancer cells to cisplatin (CDDP).

Methods

The IC50 for each drug was first determined. The cells were then pre-exposed to 40 µM of TQ for 24 h before being treated with 6 µM of cisplatin. The viability and the sub-G1 population of the 5673 cells were respectively evaluated by alamar blue assay and propidium iodide staining. RT-qPCR was also applied to analyze the expression profile of the apoptosis-related genes (Bax, Bcl-2, p53).

Results

The viability of the cells treated with the combination of TQ and CDDP was significantly decreased compared to CDDP- or TQ-treated cells. TQ at the concentration of 40 µM increased the cytotoxicity of 6 µM CDDP by 35.5%. Moreover, flow cytometry analysis indicated that TQ pre-treatment of the cells resulted in a 55.5% increase in the population of 5637 cells in the sub-G1 phase compared to cells treated with CDDP alone. The results from RT-qPCR exhibited that the exposure of the cells to both TQ and CDDP significantly elevated Bax/Bcl-2 ratio by down-regulating Bcl-2 expression.

Conclusion

TQ significantly increased the cytotoxicity of CDDP in 5637 cells and induced apoptosis by down-regulation of the Bcl-2. Therefore, TQ and CDDP might be an effective therapeutic combination for TCC bladder cancer treatment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Data Availability

Data sharing not applicable to this article as no datasets were generated or analysed during the current study.

Abbreviations

CDDP:

Cisplatin

TQ:

Thymoquinone

References

  1. Bray F et al (2021) The ever-increasing importance of cancer as a leading cause of premature death worldwide. Cancer 127(16):3029–3030

    Article  PubMed  Google Scholar 

  2. Sung H et al (2021) Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. Cancer J Clin 71(3):209–249

    Article  Google Scholar 

  3. Richters A, Aben KK, Kiemeney LA (2020) The global burden of urinary bladder cancer: an update. World J Urol 38(8):1895–1904

    Article  PubMed  Google Scholar 

  4. Rutz J et al (2020) Curcumin—a viable agent for better bladder cancer treatment. Int J Mol Sci 21(11):3761

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Patel VG, Oh WK, Galsky MD (2020) Treatment of muscle-invasive and advanced bladder cancer in 2020 CA: a cancer journal for clinicians. 70(5):404–423

  6. Anastasiadis A, de Reijke TM (2012) Best practice in the treatment of nonmuscle invasive bladder cancer. Ther Adv Urol 4(1):13–32

    Article  PubMed  PubMed Central  Google Scholar 

  7. Park BH et al (2016) Curcumin potentiates antitumor activity of cisplatin in bladder cancer cell lines via ROS-mediated activation of ERK1/2. Oncotarget 7(39):63870

    Article  PubMed  PubMed Central  Google Scholar 

  8. Dobruch J, Oszczudłowski M (2021) Bladder cancer: current challenges and future directions. Medicina 57(8):749

    Article  PubMed  PubMed Central  Google Scholar 

  9. Rassouli FB et al (2011) Investigating the enhancement of cisplatin cytotoxicity on 5637 cells by combination with mogoltacin. Toxicol In Vitro 25(2):469–474

    Article  CAS  PubMed  Google Scholar 

  10. Rodriguez RHM, Rueda OB, Ibarz L (2017) Bladder cancer: present and future. Med Clínica (English Edition) 149(10):449–455

    Article  Google Scholar 

  11. Lheureux S, Braunstein M, Oza AM (2019) Epithelial ovarian cancer: evolution of management in the era of precision medicine. Cancer J Clin 69(4):280–304

    Google Scholar 

  12. de Vries G et al (2020) Testicular cancer: determinants of cisplatin sensitivity and novel therapeutic opportunities. Cancer Treat Rev 88:102054

    Article  PubMed  Google Scholar 

  13. Kamat AM et al (2016) Bladder cancer. The Lancet 388(10061):2796–2810

    Article  Google Scholar 

  14. Kaidar-Person O, Gil Z, Billan S (2018) Precision medicine in head and neck cancer. Drug Resist Updates 40:13–16

    Article  Google Scholar 

  15. Lemjabbar-Alaoui H et al (2015) Lung cancer: Biology and treatment options. Biochim et Biophys Acta (BBA)-Reviews Cancer 1856(2):189–210

    Article  CAS  Google Scholar 

  16. Li H, Wu X, Cheng X (2016) Advances in diagnosis and treatment of metastatic cervical cancer.Journal of gynecologic oncology, 27(4)

  17. Ghosh S (2019) Cisplatin: the first metal based anticancer drug. Bioorg Chem 88:102925

    Article  CAS  PubMed  Google Scholar 

  18. Holditch SJ et al (2019) Recent advances in models, mechanisms, biomarkers, and interventions in cisplatin-induced acute kidney injury. Int J Mol Sci 20(12):3011

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Dos Santos NAG, Ferreira RS, Dos AC, Santos (2020) Overview of cisplatin-induced neurotoxicity and ototoxicity, and the protective agents. Food Chem Toxicol 136:111079

    Article  Google Scholar 

  20. Khan MA et al (2017) Thymoquinone, as an anticancer molecule: from basic research to clinical investigation. Oncotarget 8(31):51907

    Article  Google Scholar 

  21. Malik S et al (2021) Thymoquinone: a small molecule from nature with high therapeutic potential. Drug Discovery Today 26(11):2716–2725

    Article  CAS  PubMed  Google Scholar 

  22. Wang Y-M (2011) Inhibitory effects of thymoquinone on human pancreatic carcinoma orthotopically implanted in nude mice. Zhonghua yi xue za zhi 91(44):3111–3114

    CAS  PubMed  Google Scholar 

  23. Imani S et al (2017) MicroRNA-34a targets epithelial to mesenchymal transition-inducing transcription factors (EMT-TFs) and inhibits breast cancer cell migration and invasion. Oncotarget 8(13):21362

    Article  PubMed  PubMed Central  Google Scholar 

  24. Dirican A et al (2015) Novel combination of docetaxel and thymoquinone induces synergistic cytotoxicity and apoptosis in DU-145 human prostate cancer cells by modulating PI3K–AKT pathway. Clin Transl Oncol 17(2):145–151

    Article  CAS  PubMed  Google Scholar 

  25. Wilson AJ et al (2015) Thymoquinone enhances cisplatin-response through direct tumor effects in a syngeneic mouse model of ovarian cancer. J ovarian Res 8(1):1–10

    Article  CAS  Google Scholar 

  26. Gali-Muhtasib H et al (2008) Thymoquinone triggers inactivation of the stress response pathway sensor CHEK1 and contributes to apoptosis in colorectal cancer cells. Cancer Res 68(14):5609–5618

    Article  CAS  PubMed  Google Scholar 

  27. Jafri SH et al (2010) Thymoquinone and cisplatin as a therapeutic combination in lung cancer: in vitro and in vivo. J Experimental Clin Cancer Res 29(1):1–11

    Article  Google Scholar 

  28. Ng WK, Yazan LS, Ismail M (2011) Thymoquinone from Nigella sativa was more potent than cisplatin in eliminating of SiHa cells via apoptosis with down-regulation of Bcl-2 protein. Toxicol In Vitro 25(7):1392–1398

    Article  CAS  PubMed  Google Scholar 

  29. Riccardi C, Nicoletti I (2006) Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 1(3):1458–1461

    Article  CAS  PubMed  Google Scholar 

  30. Kundu J et al (2014) Thymoquinone induces apoptosis in human colon cancer HCT116 cells through inactivation of STAT3 by blocking JAK2-and src–mediated phosphorylation of EGF receptor tyrosine kinase. Oncol Rep 32(2):821–828

    Article  PubMed  Google Scholar 

  31. El-Far AH et al (2020) Thymoquinone-chemotherapeutic combinations: new regimen to combat cancer and cancer stem cells. Naunyn Schmiedebergs Arch Pharmacol 393(9):1581–1598

    Article  CAS  PubMed  Google Scholar 

  32. Cascella M et al (2017) Role of Nigella sativa and its constituent thymoquinone on chemotherapy-induced nephrotoxicity: evidences from experimental animal studies. Nutrients 9(6):625

    Article  PubMed  PubMed Central  Google Scholar 

  33. Badary OA et al (2000) The influence of thymoquinone on doxorubicin-induced hyperlipidemic nephropathy in rats. Toxicology 143(3):219–226

    Article  CAS  PubMed  Google Scholar 

  34. HADJZADEH MAR et al (2012) Effect of alcoholic extract of Nigella sativa on cisplatininduced toxicity in rat

  35. Ulu R et al (2012) Regulation of renal organic anion and cation transporters by thymoquinone in cisplatin induced kidney injury. Food Chem Toxicol 50(5):1675–1679

    Article  CAS  PubMed  Google Scholar 

  36. Badary OA (1999) Thymoquinone attenuates ifosfamide-induced Fanconi syndrome in rats and enhances its antitumor activity in mice. J Ethnopharmacol 67(2):135–142

    Article  CAS  PubMed  Google Scholar 

  37. Zhou X et al (2021) Thymoquinone suppresses the proliferation, migration and invasiveness through regulating ROS, autophagic flux and mir-877-5p in human bladder carcinoma cells. Int J Biol Sci 17(13):3456

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Zhang M et al (2020) Thymoquinone suppresses invasion and metastasis in bladder cancer cells by reversing EMT through the Wnt/β-catenin signaling pathway. Chemico-Biol Interact 320:109022

    Article  CAS  Google Scholar 

  39. Zhang G-J et al (1999) Tamoxifen-induced apoptosis in breast cancer cells relates to down-regulation of bcl-2, but not bax and bcl-XL, without alteration of p53 protein levels. 5(10):2971–2977

  40. Jehan S et al (2020) Thymoquinone selectively induces hepatocellular carcinoma cell apoptosis in synergism with clinical therapeutics and dependence of p53 status. Front Pharmacol 11:555283

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Wang Y-C et al (2022) Severe cellular stress drives apoptosis through a dual control mechanism independently of p53. Cell Death Discovery 8(1):282

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Toshiyuki M, Reed JC (1995) Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 80(2):293–299

    Article  Google Scholar 

  43. Fatfat Z, Fatfat M, Gali-Muhtasib H (2021) Therapeutic potential of thymoquinone in combination therapy against cancer and cancer stem cells. World J Clin Oncol 12(7):522

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Birjand University of Medical Sciences for supporting the project.

Author information

Authors and Affiliations

  1. Department of Medical Biotechnology, School of Medicine, Birjand University of Medical Sciences, Birjand, Iran

    Fatemeh Khodadadi, Mohsen Khorashadizadeh & Fahimeh Ghasemi

  2. Cellular and Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran

    Mohsen Khorashadizadeh & Fahimeh Ghasemi

Authors
  1. Fatemeh Khodadadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mohsen Khorashadizadeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fahimeh Ghasemi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Fahimeh Ghasemi.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Khodadadi, F., Khorashadizadeh, M. & Ghasemi, F. Thymoquinone enhanced the antitumor activity of cisplatin in human bladder cancer 5637 cells in vitro. Mol Biol Rep 50, 5767–5775 (2023). https://doi.org/10.1007/s11033-023-08472-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-023-08472-8

Keywords

Search

Navigation