Archives of Advances in Biosciences

Vol. 17 No. 1 (2026)

Suppression of CircPVT1 Potentiates 5-Fluorouracil-Driven Programmed Cell Death in SK-MEL-37 Malignant Melanoma Cells: Role of the miR-203a-3p/FGF2 Regulatory Cascade

Archives of Advances in Biosciences, Vol. 17 No. 1 (2026), 28 Ordibehesht 2026 , Page 1-13
https://doi.org/10.22037/aab.v17i1.52479

Abstract

Background and Aim: The circular RNA, known as CircPVT1, functions as an oncogenic molecule across several.

Methods: SK-MEL-37 cultures were assigned to five experimental conditions: untreated controls, non-targeting scrambled siRNA (40 nM), CircPVT1-directed siRNA (40 nM), 5-FU alone (500 µM), and the siRNA/5-FU combination. Transfection employed TurboFect reagent. Following a 48-hour exposure, viability was evaluated through MTT colorimetry. Apoptotic rates were determined via Annexin V-FITC/propidium iodide dual labeling coupled with flow cytometry. Quantitative real-time PCR measured transcript levels of CircPVT1, miR-203a-3p, FGF2, Bax, Bcl-2, and Caspase-3. Bioinformatic tools (circInteractome, TargetScan, miRDB) predicted possible interactions within the CircPVT1/miR-203a-3p/FGF2 network.

Results: In silico analysis revealed a potential miR-203a-3p docking site inside CircPVT1 and identified FGF2 as a genuine miR-203a-3p target carrying a conserved 3′UTR recognition sequence. MTT data showed that anti-CircPVT1 siRNA lowered viability to 48.3% of control values, whereas 5-FU alone dropped viability to 44.2%. The combined regimen produced the strongest reduction, yielding 28.6% viability (p<0.001 versus controls; p<0.01 versus each monotherapy). Flow cytometry indicated that the combination elevated late-stage apoptosis to 24.3%, compared with 17.9% for siRNA alone and 15.7% for 5-FU alone (p<0.001). qRT-PCR revealed that the combination upregulated miR-203a-3p by 5.76-fold and Bax by 2.36-fold, while downregulating CircPVT1 to 0.22-fold, FGF2 to 0.11-fold, and Bcl-2 to 0.18-fold. The Bax/Bcl-2 ratio rose from 1.0 (controls) to 13.1 in the combination arm.

Conclusion: Our findings indicate that CircPVT1 knockdown markedly boosts 5-FU-provoked apoptosis in SK-MEL-37 melanoma cells, an effect likely mediated through restored miR-203a-3p expression and subsequent FGF2 pathway inhibition.

Keywords:
  • Melanoma
  • CircPVT1
  • miR 203a 3p
  • FGF2
  • 5 Fluorouracil
  • Apoptosis
  • Chemosensitization

How to Cite

Yousefian Naeini, Z., Hashemi, M., Heshmati , M., Jebali, A., & Entezari, M. (2026). Suppression of CircPVT1 Potentiates 5-Fluorouracil-Driven Programmed Cell Death in SK-MEL-37 Malignant Melanoma Cells: Role of the miR-203a-3p/FGF2 Regulatory Cascade. Archives of Advances in Biosciences, 17(1), 1–13. https://doi.org/10.22037/aab.v17i1.52479

References

1. Schadendorf D, van Akkooi ACJ, Berking C, et al. Melanoma. Lancet. 2020;395(10236):1835-44.) DOI: 10.1016/S0140-6736(18)31559-9( (PMID)

2. Arnold M, Singh D, Laversanne M, et al. Global burden of cutaneous melanoma in 2020 and projections to 2040. JAMA Dermatol. 2022;158(5):495-503. (DOI: 10.1001/jamadermatol.2022.0160) (PMID)

3. Saginala K, Barsouk A, Aluru JS, Rawla P, Barsouk A. Epidemiology of melanoma. Med Sci. 2021;9(4):63. (DOI: 10.3390/medsci9040063) (PMID)

4. Luke JJ, Flaherty KT, Ribas A, Long GV. Targeted agents and immunotherapies: optimizing outcomes in melanoma. Nat Rev Clin Oncol. 2017;14(8):463-82. (DOI: 10.1038/nrclinonc.2017.43) (PMID)

5. Robert C, Grob JJ, Stroyakovskiy D, et al. Five-year outcomes with dabrafenib plus trametinib in metastatic melanoma. N Engl J Med. 2019;381(7):626-36. (DOI: 10.1056/NEJMoa1904059) (PMID)

6. Larkin J, Chiarion-Sileni V, Gonzalez R, et al. Five-year survival with combined nivolumab and ipilimumab in advanced melanoma. N Engl J Med. 2019;381(16):1535-46. (DOI: 10.1056/NEJMoa1910836) (PMID)

7. Wolchok JD, Chiarion-Sileni V, Gonzalez R, et al. Long-term outcomes with nivolumab plus ipilimumab or nivolumab alone in advanced melanoma. J Clin Oncol. 2022;40(2):127-37. (DOI: 10.1200/JCO.21.02229) (PMID)

8. Shi H, Hugo W, Kong X, et al. Acquired resistance and clonal evolution in melanoma during BRAF inhibitor therapy. Cancer Discov. 2014;4(1):80-93. (DOI: 10.1158/2159-8290.CD-13-0642) (PMID)

9. Jenkins RW, Barbie DA, Flaherty KT. Mechanisms of resistance to immune checkpoint inhibitors. Br J Cancer. 2018;118(1):9-16. (DOI: 10.1038/bjc.2017.434) (PMID)

10. Sharma P, Hu-Lieskovan S, Wargo JA, Ribas A. Primary adaptive and acquired resistance to cancer immunotherapy. Cell. 2017;168(4):707-23. (DOI: 10.1016/j.cell.2017.01.017) (PMID)

11. Domingues B, Lopes JM, Soares P, Populo H. Melanoma treatment in review. Immunotargets Ther. 2018;7:35-49. (DOI: 10.2147/ITT.S134842) (PMID)

12. Longley DB, Harkin DP, Johnston PG. 5-Fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer. 2003;3(5):330-8. (DOI: 10.1038/nrc1074) (PMID)

13. Najem A, Krayem M, Perdrix A, et al. New drug combination strategies in melanoma: current status and future directions. Anticancer Res. 2017;37(11):5941-53. (DOI: 10.21873/anticanres.12041) (PMID)

14. Yun F, Wu N, Yi X, Zhang X, Feng Y, Ni Q, et al. Nod2 reduces the chemoresistance of melanoma by inhibiting the tyms/plk1 signaling axis. Cell Death Dis. 2024;15(10):720-25. (DOI: 10.1038/s41419-024-07104-8)

15. Kristensen LS, Andersen MS, Stagsted LVW, Ebbesen KK, Hansen TB, Kjems J. The biogenesis biology and characterization of circular rnas. Nat Rev Genet. 2019;20(11):675-91. (DOI: 10.1038/s41576-019-0158-7) (PMID)

16. Jeck WR, Sharpless NE. Detecting and characterizing circular rnas. Nat Biotechnol. 2014;32(5):453-61. (DOI: 10.1038/nbt.2890) (PMID)

17. Chen LL. The expanding regulatory mechanisms and cellular functions of circular rnas. Nat Rev Mol Cell Biol. 2020;21(8):475-90. (LINK)

18. Hansen TB, Jensen TI, Clausen BH, et al. Natural rna circles function as efficient microrna sponges. Nature. 2013;495(7441):384-8. (DOI: 10.1038/nature11993) (PMID)

19. Vo JN, Cieslik M, Zhang Y, et al. The landscape of circular rna in cancer. Cell. 2019;176(4):869-81. Link

20. Patop IL, Kadener S. circrnas in cancer. Curr Opin Genet Dev. 2018;48:121-127. (DOI: 10.1016/j.gde.2017.11.007)

21. Wang YY, Duan H, Wang S, Quan YJ, Huang JH, Guo ZC. Elevated circular rna pvt1 promotes eutopic endometrial cell proliferation and invasion of adenomyosis via mir-145/talin1 axis. Biomed Res Int. 2021;2021:8868700. (DOI: 10.1155/2021/8868700) (PMID)

22. Li X, Liu CX, Xue W, et al. Circular rna circpvt1 promotes proliferation and invasion through sponging mir-125b in gastric cancer. Cell Physiol Biochem. 2018;48(6):2322-33. (DOI: 10.1159/000495876)

23. Qin S, Zhao Y, Zhang L, Lim G, Lin H, Zhang X. Circular rnA pvt1 acts as a competing endogenous rna for mir-497 in promoting non-small cell lung cancer progression. Biomed Pharmacother. 2019;111:244-250. (DOI: 10.1016/j.biopha.2018.12.007) (PMID)

24. Ma X, Chen J, Zhou C. Circular rna pvt1 as a potential biomarker for human cancers: a systematic review and meta-analysis. Int J Biol Markers. 2025;40(3):158-65. (DOI: 10.1177/03936155251362862) (PMID)

25. Wang S, Li W, Yang L, Yuan J, Wang L, Li N, et al. Circpvt1 facilitates the progression of oral squamous cell carcinoma by regulating mir-143-3p/slc7a11 axis through mapk signaling pathway. Funct Integr Genomics. 2022;22(5):891-903.(DOI: 10.1007/s10142-022-00865-5) (PMID)

26. Zhang M, He A, Xu Z, Chen M, Zhang H, Mao W. Circpvt1 regulates emt and induces macrophage polarization to promotes the progression of renal cell carcinoma. Front Immunol. 2026;17:1760058. (DOI: 10.3389/fimmu.2026.1760058) (PMID)

27. Bartel DP. Metazoan micrornas. Cell. 2018;173(1):20-51. (DOI: 10.1016/j.cell.2018.03.006) (PMID)

28. Obayashi M, Yoshida M, Tsunematsu T, Ogawa I, Sasahira T, Kuniyasu H, et al. microrna-203 suppresses invasion and epithelial-mesenchymal transition induction via targeting nuak1 in head and neck cancer. Oncotarget. 2016;7(7):8223. (DOI: 10.18632/oncotarget.6972) (PMID)

29. Noguchi S, Mori T, Otsuka Y, Yamada N, Yasui Y, Iwasaki J, et al. Anti-oncogenic microrna-203 induces senescence by targeting e2ff3 protein in human melanoma cells. J Biol Chem. 2012;287(15):11769-77. (DOI: 10.1074/jbc.M111.325027) (PMID)

30. Lohcharoenkal W, Mahapatra KD, Pasquali L, Crudden C, Kular L, Ulum YZ, et al. Genome-wide screen for micrornas reveals a role for mir-203 in melanoma metastasis. J Inves Dermatol. 2018;138(4):882-92. (DOI: 10.1016/j.jid.2017.09.049) (PMID)

31. Fu XF, Li Y, Wang Y, et al. Mir-203 inhibits the proliferation invasion and migration of pancreatic cancer cells by down-regulating fibroblast growth factor 2. Cancer Cell Int. 2020;20:166. (DOI: 10.3892/ol.2021.12887) (PMID)

32. Presta M, Chiodelli P, Giacomini A, Rusnati M, Ronca R. Fibroblast growth factor/fibroblast growth factor receptor system in angiogenesis. Cytokine Growth Factor Rev. 2021;58:47-57. (DOI: 10.1016/j.cytogfr.2005.01.004) (PMID)

33. Czyz M. Fibroblast growth factor receptor signaling in skin cancers. Cells. 2019;8(6):540. (DOI: 10.3390/cells8060540) (PMID)

34. Tait SW, Green DR. Mitochondria and cell death: outer membrane permeabilization and beyond. Nat Rev Mol Cell Biol. 2010;11(9):621-32. (DOI: 10.1038/nrm2952) (PMID)

35. Galluzzi L, Vitale I, Aaronson SA, et al. Molecular mechanisms of cell death: recommendations of the nomenclature committee on cell death 2018. Cell Death Differ. 2018;25(3):486-541. (DOI: 10.1038/s41418-017-0012-4) (PMID)

36. Hata AN, Engelman JA, Faber AC. The bcl2 family: key mediators of the apoptotic response to targeted anticancer therapeutics. Cancer Discov. 2015;5(5):475-87. (DOI: 10.1158/2159-8290.CD-15-0011)

37. Liu Z, Wang Y, Wang Y, Zhang Y. Exosomal circrnas in hepatocellular carcinoma: implications for the development and therapeutic resistance of hepatocellular carcinoma. Int J Oncol. 2025;68(2):26. (DOI: 10.3892/ijo.2025.5839) (PMID)

38. Ma S, Kong S, Wang F, Ju S. Circrnas: biogenesis functions and role in drug-resistant tumours. Molecular Cancer. 2020 ;19(1):119. (DOI: 10.1186/s12943-020-01253-y)

39. Davidson BL, McCray PB Jr. Current prospects for rna interference-based therapies. Nat Rev Genet. 2011;12(5):329-40. (DOI: 10.1038/nrg2968) (PMID)

40. Thomson DW, Dinger ME. Endogenous microrna sponges: evidence and controversy. Nat Rev Genet. 2016;17(5):272-83. (DOI: 10.1038/nrg.2016.20) (PMID)

41. Li J, Sun D, Pu W, Wang J, Peng Y. Circular rnas in cancer: biogenesis function and clinical significance. Trends Cancer. 2020;6(4):319-36. (DOI: 10.1016/j.trecan.2020.01.012)

42. Liu CX, Chen LL. Circular rnas: characterization, cellular roles and applications. Cell. 2022;185(12):2016-34. (DOI: 10.1016/j.cell.2022.04.021)

43. Palcau AC, Canu V, Donzelli S, Strano S, Pulito C, Blandino G. Circpvt1: a pivotal circular node intersecting long non-coding-pvt1 and c-myc oncogenic signals. Molecular Cancer. 2022;21(1):33. (DOI: 10.1186/s12943-022-01514-y) (PMID)

44. Xiao M, Feng Y, Liu C, Zhang Z. Prognostic values of long noncoding rna pvt 1 in various carcinomas: An updated systematic review and meta‐analysis. Cell proliferation. 2018;51(6):e12519.(DOI: 10.1111/cpr.12519) (PMID)

45. Melixetian M, Pelicci PG, Lanfrancone L. Regulation of lncrnas in melanoma and their functional roles in the metastatic process. Cells. 2022;11(3):577. (DOI: 10.3390/cells11030577) (PMID)

46. Vogel C, Marcotte EM. Insights into the regulation of protein abundance from proteomic and transcriptomic analyses. Nat Rev Genet. 2012;13(4):227-32. (DOI: 10.1038/nrg3185) (PMID)

47. Liu Y, Beyer A, Aebersold R. On the dependency of cellular protein levels on mRNA abundance. Cell. 2016;165(3):535-50. (DOI: 10.1016/j.cell.2016.03.014) (PMID)

48. Porter AG, Jänicke RU. Emerging roles of caspase-3 in apoptosis. Cell Death Differ. 1999;6(2):99-104. (DOI: 10.1038/sj.cdd.4400476) (PMID)

49. Turner N, Grose R. Fibroblast growth factor signalling: from development to cancer. Nat Rev Cancer. 2010;10(2):116-29. (DOI: 10.1038/nrc2780) (PMID)

50. Diasio RB, Harris BE. Clinical pharmacology of 5-fluorouracil. Clin Pharmacokinet. 1989;16(4):215-37. (DOI: 10.2165/00003088-198916040-00002) (PMID)

51. Longley DB, Harkin DP, Johnston PG. 5-Fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer. 2003;3(5):330-8. (DOI: 10.1038/nrc1074) (PMID)

52. Peters GJ, Backus HHJ, Freemantle S, et al. Induction of thymidylate synthase as a 5-fluorouracil resistance mechanism. Biochim Biophys Acta. 2002;1587(2-3):194-205. (DOI: 10.1016/S0925-4439(02)00074-9) (PMID)

53. Mader RM, Müller M, Steger GG. Resistance to 5-fluorouracil: mechanisms and clinical implications.

Cancer Treat Rev. 1998;24(1):1-22. (DOI: 10.1016/S0305-7372(98)90002-1) (PMID)

54. Gottesman MM. Mechanisms of cancer drug resistance.

Annu Rev Med. 2002;53:615-27. (DOI: 10.1146/annurev.med.53.082901.103929) (PMID)

55. Longley DB, Johnston PG. Molecular mechanisms of drug resistance. J Pathol. 2005;205(2):275-92. (DOI: 10.1002/path.1704) (PMID)

56. Ekwall B. Screening of toxic compounds in mammalian cell cultures. Toxicology. 1999;138(1):1-10. (DOI: 10.1016/S0300-483X(99)00068-4) (PMID)

57. McGivan JD, Hayes JD. The role of glutathione in drug resistance. Adv Cancer Res. 1989;52:249-81. (DOI: 10.1016/S0065-230X(08)60312-5) (PMID)

58. Hanahan D. Hallmarks of Cancer: New Dimensions.

Cancer Discov. 2022;12(1):31-46. (DOI: 10.1158/2159-8290.CD-21-1059) (PMID)

59. Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646-74. (DOI: 10.1016/j.cell.2011.02.013) (PMID)

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