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Regulatory Effects of WRAP53 on Radiosensitivity of Laryngeal Squamous Cell Carcinoma Cells

  • Qiu, Hui (Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Cancer Clinical Study Center of Hubei Provinces, Key Laboratory of Tumor Biological Behavior of Hubei Province) ;
  • Zhao, De-Ying (Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Cancer Clinical Study Center of Hubei Provinces, Key Laboratory of Tumor Biological Behavior of Hubei Province) ;
  • Yuan, Li-Mei (Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Cancer Clinical Study Center of Hubei Provinces, Key Laboratory of Tumor Biological Behavior of Hubei Province) ;
  • Zhang, Gong (Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Cancer Clinical Study Center of Hubei Provinces, Key Laboratory of Tumor Biological Behavior of Hubei Province) ;
  • Xie, Cong-Hua (Department of Radiation & Medical Oncology, Zhongnan Hospital of Wuhan University, Cancer Clinical Study Center of Hubei Provinces, Key Laboratory of Tumor Biological Behavior of Hubei Province)
  • Published : 2015.04.14

Abstract

Background: Telomere length is closely associated with cellular radiosensitivity and WRAP53 is required for telomere addition by telomerase. In this research we assessed radiosensitivity of laryngeal squamous cell carcinoma Hep-2 cell lines after WRAP53 inhibition, and analyzed the molecular mechanisms. Materials and Methods: phWRAP53-siRNA and pNeg-siRNA were constructed and transfected into Hep-2 cells with lipofectamine. Expression of WRAP53 was analyzed by RT-PCR and Western-blottin, radiosensitivity of Hep-2 cells was assessed colony formation assay, and the relative length of telomeres was measured by QPCR. Results: The data revealed that the plasmid of phWRAP53-siRNA was constructed successfully, and the mRNA and protein levels of WRAP53 were both obviously reduced in the Hep-2 cell line transfected with phWRAP53-siRNA. After Hep-2 cells were irradiated with X-rays, the $D_0$ and $SF_2$ were 2.481 and 0.472, respectively, in the phWRAP53-siRNA group, much lower than in the control group ($D_0$ and $SF_2$ of 3.213 and 0.592) (P<0.01). The relative telomere length in the phWRAP53-siRNA group was $0.185{\pm}0.01$, much lower than in the untreated group ($0.523{\pm}0.06$) and the control group ($0.435{\pm}0.01$). Conclusions: Decreasing the expression of WRAP53 using RNA interference technique can enhance the radiosensitivity of Hep-2 cell lines by influencing the telomere length. WRAP53 is expected to be a new target to regulate the radiosensitization of tumor cells.

Keywords

WRAP53;Hep-2;laryngeal squamous cell carcinoma;radiosensitivity;telomere length

References

  1. Arshad H, Jayaprakash V, Gupta V, et al (2014). Survival differences between organ preservation surgery and definitive radiotherapy in early supraglottic squamous cell carcinoma. Otolaryngol Head Neck Surg, 150, 237-44. https://doi.org/10.1177/0194599813512783
  2. Cawthon RM (2002). Telomere measurement by quantitative PCR. Nucleic Acids Res, 30, 47. https://doi.org/10.1093/nar/30.1.47
  3. Fairlie J, Harrington L (2015). Enforced telomere elongation increases the sensitivity of human tumour cells to ionizing radiation. DNA Repair, 25, 54-9. https://doi.org/10.1016/j.dnarep.2014.11.005
  4. Freund A, Zhong FL, Venteicher AS, et al (2014). Proteostatic control of telomerase function through TRiC-mediated folding of TCAB1. Cell, 159, 1389-403. https://doi.org/10.1016/j.cell.2014.10.059
  5. Hu Y, Bobb D, Lu Y, et al (2014). Effect of telomerase inhibition on preclinical models of malignant rhabdoid tumor. Cancer Genet, 207, 403-11. https://doi.org/10.1016/j.cancergen.2014.09.002
  6. Laster BH, Isaacson C, Perets E, et al (2015). Keeping those telomeres short! an innovative intratumoral long-term drug delivery system. J Cancer Res Clin Oncol, 141, 23-34. https://doi.org/10.1007/s00432-014-1747-7
  7. Mahmoudi S, Henriksson S, Farnebo L (2011). WRAP53 promotes cancer cell survival and is a potential target for cancer therapy. Cell Death Dis, 2, 114. https://doi.org/10.1038/cddis.2010.90
  8. Megwalu UC, Sikora AG (2014). Survival outcomes in advanced laryngeal cancer. JAMA Otolaryngol Head Neck Surg, 140, 855-60. https://doi.org/10.1001/jamaoto.2014.1671
  9. Mok G, Gauthier I, Jiang H, et al (2014). Outcomes of intensitymodulated radiotherapy versus conventional radiotherapy for hypopharyngeal cancer. Head Neck, [Epub ahead of print].
  10. Saldana-Meyer R, Gonzalez-Buendia E, Guerrero G, et al (2014). CTCF regulates the human p53 gene through direct interaction with its natural antisense transcript, Wrap53. Genes Dev, 28, 723-34. https://doi.org/10.1101/gad.236869.113
  11. Sedaie Bonab A, Pouladi N, Hosseinpourfeizi MA, et al (2014). Single-strand conformational polymorphism analysis of a common single nucleotide variation in WRAP53 gene, rs2287499, and evaluating its association in relation to breast cancer risk and prognosis among Iranian-Azeri population. Med Oncol, 31, 168. https://doi.org/10.1007/s12032-014-0168-4
  12. Stern JL, Zyner KG, Pickett HA, et al (2012). Telomerase recruitment requires both TCAB1 and Cajal bodies independently. Mol Cell Biol, 32, 2384-95. https://doi.org/10.1128/MCB.00379-12
  13. Sun CK, Luo XB, Gou YP, et al (2014). TCAB1: a potential target for diagnosis and therapy of head and neck carcinomas. Mol Cancer, 13, 180. https://doi.org/10.1186/1476-4598-13-180
  14. Venteicher AS, Artandi SE (2009). TCAB1: driving telomerase to Cajal bodies. Cell Cycle, 8, 1329-31. https://doi.org/10.4161/cc.8.9.8288
  15. Zhong F, Savage SA, Shkreli M, et al (2011). Disruption of telomerase trafficking by TCAB1 mutation causes dyskeratosis congenita. Genes Dev, 25, 11-6. https://doi.org/10.1101/gad.2006411

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