Role of The circ-HIPK3, circ-PVT1, miR-25, and miR-149 in Response of Breast Cancer Cells to Ionizing Radiation

Document Type : Original Article

Authors

Department of Medical Genetics, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

Abstract

Objective: Determining cellular radiosensitivity of breast cancer (BC) patients through molecular markers before
radiation therapy (RT) allows accurate prediction of individual’s response to radiation. The aim of this study was
therefore to investigate the potential role of epigenetic biomarkers in breast cancer cellular radiosensitivity.
Materials and Methods: In this experimental study, we treated two BC cell lines, MDA-MB 231 and MCF-7, with doses
of 2, 4, and 8Gy of irradiation for 24 and 48 hours. Expression levels of circ-HIPK3, circ-PVT1, miR-25, and miR-
149 were quantified using quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Significance of the
observations was statistically verified using one-way ANOVA with a significance level of P<0.05. Annexin V-FITC/PI
binding assay was utilized to measure cellular apoptosis.
Results: The rate of cell apoptosis was significantly higher in MCF-7 cells compared to MDA-MB-231 cells at doses of
4Gy and 8Gy (P=0.013 and P=0.004, respectively). RNA expression analysis showed that circ-HIPK3 was increased in
the MDA-MB-231 cell line compared to the MCF-7 cell line after exposure to 8Gy for 48 hours. Expression of circ-PVT1
was found to be higher in MDA-MB-231 cells compared to MCF-7 cells after exposure to 8Gy for 24 hours, likewise
after exposure to 4Gy and 8Gy for 48 hours. After exposing 8Gy, expression of miR-25 was increased in MDA-MB-231
cells compared to MCF-7 cells at 24 and 48 hours. After exposing 8Gy dose, expression of miR-149 was increased in
MCF-7 cells compared to MDA-MB-231 cells at 24 and 48 hours.
Conclusion: circ-HIPK3, circ-PVT1, and miR-25 played crucial roles in the mechanisms of radioresistance in breast
cancer. Additionally, miR-149 was involved in regulating cellular radiosensitivity. Therefore, these factors provided
predictive information about a tumor’s radiosensitivity or its response to treatment, which could be valuable in
personalizing radiation dosage.

Keywords

Main Subjects

References

  1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011; 61(2): 69-90.
  2. Delaney G, Jacob S, Featherstone C, Barton M. The role of radiotherapy in cancer treatment: estimating optimal utilization from a review of evidence-based clinical guidelines. Cancer. 2005; 104(6): 1129-1137.
  3. Early Breast Cancer Trialists’ Collaborative Group (EBCTCG); Darby S, McGale P, Correa C, Taylor C, Arriagada R, et al. Effect

of radiotherapy after breast-conserving surgery on 10-year recurrence and 15-year breast cancer death: meta-analysis of individual patient data for 10,801 women in 17 randomised trials. Lancet. 2011; 378(9804): 1707-1716.

  1. Liu X, Li F, Huang Q, Zhang Z, Zhou L, Deng Y, et al. Self-inflicted DNA double-strand breaks sustain tumorigenicity and stemness ofcancer cells. Cell Res. 2017; 27(6): 764-783.
  2. Jackson SP, Bartek J. The DNA-damage response in human biology and disease. Nature. 2009; 461(7267): 1071-1078.
  3. Xu L, Lyu M, Yang S, Zhang J, Yu D. CircRNA expression profiles of breast cancer and construction of a circRNA-miRNA-mRNA network. Sci Rep. 2022; 12(1): 17765.
  4. Zheng Q, Bao C, Guo W, Li S, Chen J, Chen B, et al. Circular RNA profiling reveals an abundant circHIPK3 that regulates cell growth by sponging multiple miRNAs. Nat Commun. 2016; 7: 11215.
  5. Kong D, Shen D, Liu Z, Zhang J, Zhang J, Geng C. Circ_0008500 Knockdown improves radiosensitivity and inhibits tumorigenesis in breast cancer through the miR-758-3p/PFN2 axis. J Mammary Gland Biol Neoplasia. 2022; 27(1): 37-52.
  6. Hu F, Peng Y, Fan X, Zhang X, Jin Z. Circular RNAs: implications of signaling pathways and bioinformatics in human cancer. Cancer Biol Med. 2023; 20(2): 104-128.
  7. Zhang ZH, Wang Y, Zhang Y, Zheng SF, Feng T, Tian X, et al. The function and mechanisms of action of circular RNAs in urologic cancer. Mol Cancer. 2023; 22(1): 61.
  8. Attwaters M. In vivo RNA base editing with circular RNAs. Nat Rev Genet. 2022; 23(4): 196-197.
  9. He J, Xie Q, Xu H, Li J, Li Y. Circular RNAs and cancer. Cancer Lett. 2017; 396: 138-144.
  10. Nielsen AF, Bindereif A, Bozzoni I, Hanan M, Hansen TB, Irimia M, et al. Best practice standards for circular RNA research. Nat Methods. 2022; 19(10): 1208-1220.
  11. Zheng Q, Bao C, Guo W, Li S, Chen J, Chen B, et al. Circular RNA profiling reveals an abundant circHIPK3 that regulates cell growth by sponging multiple miRNAs. Nat Commun. 2016; 7: 11215.
  12. Zeng K, Chen X, Xu M, Liu X, Hu X, Xu T, et al. CircHIPK3 promotes colorectal cancer growth and metastasis by sponging miR- 7. Cell Death Dis. 2018; 9(4): 417.
  13. Liu Z, Guo S, Sun H, Bai Y, Song Z, Liu X. Circular RNA CircHIPK3 elevates CCND2 expression and promotes cell proliferation and invasion through miR-124 in glioma. Front Genet. 2020; 11: 1013.
  14. Chen D, Lu X, Yang F, Xing N. Circular RNA circHIPK3 promotes cell proliferation and invasion of prostate cancer by sponging miR- 193a-3p and regulating MCL1 expression. Cancer Manag Res. 2019; 11: 1415-1423.
  15. Zeng K, Chen X, Xu M, Liu X, Hu X, Xu T, et al. CircHIPK3 promotes colorectal cancer growth and metastasis by sponging miR- 7. Cell Death Dis. 2018; 9(4): 417.
  16. Lu H, Han X, Ren J, Ren K, Li Z, Sun Z. Circular RNA HIPK3 induces cell proliferation and inhibits apoptosis in non-small cell lung cancer through sponging miR-149. Cancer Biol Ther. 2020; 21(2): 113-121.
  17. Luo N, Liu S, Li X, Hu Y, Zhang K. Circular RNA circHIPK3 promotes breast cancer progression via sponging MiR-326. Cell Cycle. 2021; 20(13): 1320-1333.
  18. Ghafouri-Fard S, Khoshbakht T, Taheri M, Jamali E. A concise review on the role of CircPVT1 in tumorigenesis, drug sensitivity, and cancer prognosis. Front Oncol. 2021; 11: 762960.
  19. Li M, Chi C, Zhou L, Chen Y, Tang X. Circular PVT1 regulates cell proliferation and invasion via miR-149-5p/FOXM1 axis in ovarian cancer. J Cancer. 2021; 12(2): 611-621.
  20. Shi LP, Guo HL, Su YB, Zheng ZH, Liu JR, Lai SH. MicroRNA-149 sensitizes colorectal cancer to radiotherapy by downregulating human epididymis protein 4. Am J Cancer Res. 2018; 8(1): 30-38.
  21. Zhao T, Meng W, Chin Y, Gao L, Yang X, Sun S, et al. Identification of miR253p as a tumor biomarker: regulation of cellular functions via TOB1 in breast cancer. Mol Med Rep. 2021; 23(6): 406.
  22. He Z, Liu Y, Xiao B, Qian X. miR-25 modulates NSCLC cell radiosensitivity through directly inhibiting BTG2 expression. Biochem Biophys Res Commun. 2015; 457(3): 235-241.
  23. Barnett GC, Wilkinson JS, Moody AM, Wilson CB, Twyman N, Wishart GC, et al. The Cambridge breast intensity-modulated radiotherapy trial: patient- and treatment-related factors that influence late toxicity. Clin Oncol (R Coll Radiol). 2011; 23(10): 662-673.
  24. Tipatet KS, Davison-Gates L, Tewes TJ, Fiagbedzi EK, Elfick A, Neu B, et al. Detection of acquired radioresistance in breast cancer cell lines using Raman spectroscopy and machine learning. Analyst. 2021; 146(11): 3709-3716.
  25. Dunne AL, Price ME, Mothersill C, McKeown SR, Robson T, Hirst DG. Relationship between clonogenic radiosensitivity, radiationinduced apoptosis and DNA damage/repair in human colon cancer cells. Br J Cancer. 2003; 89(12): 2277-2283.
  26. Dayal R, Singh A, Pandey A, Mishra KP. Reactive oxygen species as mediator of tumor radiosensitivity. J Cancer Res Ther. 2014; 10(4): 811-818.
  27. Wang W, Luo YP. MicroRNAs in breast cancer: oncogene and tumor suppressors with clinical potential. J Zhejiang Univ Sci B. 2015; 16(1): 18-31.
  28. Li R, Wang X, Zhu C, Wang K. lncRNA PVT1: a novel oncogene in multiple cancers. Cell Mol Biol Lett. 2022; 27(1): 84.
  29. Wang W, Nag SA, Zhang R. Targeting the NFκB signaling pathways for breast cancer prevention and therapy. Curr Med Chem. 2015; 22(2): 264-289.
  30. Li R, Wang X, Zhu C, Wang K. lncRNA PVT1: a novel oncogene in multiple cancers. Cell Mol Biol Lett. 2022; 27(1): 84.
  31. Wang J, Huang K, Shi L, Zhang Q, Zhang S. CircPVT1 promoted the progression of breast cancer by regulating MiR-29a-3p-mediated AGR2-HIF-1α pathway. Cancer Manag Res. 2020; 12: 11477-11490.
  32. Li H, Zhu X, Zhang J, Shi J. MicroRNA-25 inhibits high glucoseinduced apoptosis in renal tubular epithelial cells via PTEN/AKT pathway. Biomed Pharmacother. 2017; 96: 471-479.
  33. Temiz E, Koyuncu İ, Sahin E. CCT3 suppression prompts apoptotic machinery through oxidative stress and energy deprivation in breast and prostate cancers. Free Radic Biol Med. 2021; 165: 88-99.

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