Tumor Biology

, Volume 35, Issue 6, pp 5971–5983 | Cite as

Deregulation of base excision repair gene expression and enhanced proliferation in head and neck squamous cell carcinoma

  • Ishrat Mahjabeen
  • Kashif Ali
  • Xiaofeng Zhou
  • Mahmood Akhtar Kayani
Research Article


Defects in the DNA damage repair pathway contribute to cancer. The major pathway for oxidative DNA damage repair is base excision repair (BER). Although BER pathway genes (OGG1, APEX1 and XRCC1) have been investigated in a number of cancers, our knowledge on the prognostic significance of these genes and their role in head and neck squamous cell carcinoma is limited. Protein levels of OGG1, APEX1 and XRCC1 and a proliferation marker, Ki-67, were examined by immunohistochemical analysis, in a cohort of 50 HNSCC patients. Significant downregulation of OGG1 (p < 0.04) and XRCC1 (p < 0.05) was observed in poorly differentiated HNSCC compared to mod–well-differentiated cases. Significant upregulation of APEX1 (p < 0.05) and Ki-67 (p < 0.05) was observed in poorly differentiated HNSCC compared to mod-well-differentiated cases. Significant correlation was observed between XRCC1 and OGG1 (r = 0.33, p < 0.02). Inverse correlations were observed between OGG1 and Ki-67 (r = −0.377, p < 0.005), between APEX1 and XRCC1 (r = −0.435, p < 0.002) and between OGG1 and APEX1 (r = −0.34, p < 0.02) in HNSCC. To confirm our observations, we examined BER pathway genes and a proliferation marker, Ki-67, expression at the mRNA level on 50 head and neck squamous cell carcinoma (HNSCC) and 50 normal control samples by quantitative real-time polymerase chain reaction. Significant downregulation was observed in case of OGG1 (p < 0.04) and XRCC1 (p < 0.02), while significant upregulation was observed in case of APEX1 (p < 0.01) and Ki-67 (p < 0.03) in HNSCC tissue samples compared to controls. Our data suggested that deregulation of base excision repair pathway genes, such as OGG1, APEX1 and XRCC1, combined with overexpression of Ki-67, a marker for excessive proliferation, may contribute to progression of HNSCC in Pakistani population.


HNSCC BER pathway gene Proliferation marker Immunohistochemistry 



This study was supported by grants from the Higher Education Commission (HEC) of Pakistan, the COMSATS Institute of Information Technology (CIIT), Islamabad, and the NIH PHS grant (CA139596).

Conflicts of interests



  1. 1.
    Spitz M, Wei Q, Dong Q, Amos CI, Wu X. Genetic susceptibility to lung cancer: the role of DNA damage and repair. Cancer Epidemiol Biomarkers Prev. 2003;12:689–98.PubMedGoogle Scholar
  2. 2.
    Tudek B. Base excision repair modulation as a risk factor for human cancers. Mol Asp Med. 2007;28:258–75.CrossRefGoogle Scholar
  3. 3.
    Mitra S, Izumi T, Boldogh I, Bhakat KK, Hill JW, Hazra TK. Choreography of oxidative damage repair in mammalian genomes. Free Radic Biol Med. 2002;33:15–28.CrossRefPubMedGoogle Scholar
  4. 4.
    Kohno T, Shinmura K, Tosaka M, Tani M, Kim SR, Sugimura H, et al. Genetic polymorphisms and altered splicing of the hOGG1 gene, that is involved in the repair of 8-hydroxyguanine in damaged DNA. Oncogene. 1998;16:3219–25.CrossRefPubMedGoogle Scholar
  5. 5.
    Campalans A, Marsin S, Nakabeppu Y, O'connor TR, Boiteux S, Radicella JP. XRCC1 interactions with multiple DNA glycosylases: a model for its recruitment to base excision repair. DNA Repair. 2005;4:826–35.CrossRefPubMedGoogle Scholar
  6. 6.
    Fan CY, Liu KL, Huang HY, Barnes EL, Swalsky PA, Bakker A, et al. XRCC1 co-localizes and physically interacts with PCNA. Nucleic Acids Res. 2004;32:2193–201.PubMedCentralCrossRefPubMedGoogle Scholar
  7. 7.
    Kumar A, Pant MC, Singh SH, Khandelwal S. Reduced expression of DNA repair genes (XRCC1, XPD, and OGG1) in squamous cell carcinoma of head and neck in North India. Tumor Biol. 2012;33(1):111–9.CrossRefGoogle Scholar
  8. 8.
    Fortini P, Pascucci B, Parlanti E, D’Errico M, Simonelli V, Dogliotti E. The base excision repair: mechanisms and its relevance for cancer susceptibility. Biochimie. 2003;85:1053–71.CrossRefPubMedGoogle Scholar
  9. 9.
    Parsons JL, Dianova I, Dianov GL. APE1 is the major 3’-phosphoglycolate activity in human cell extracts. Nucleic Acids Res. 2004;32:3531–6.PubMedCentralCrossRefPubMedGoogle Scholar
  10. 10.
    Vidal AE, Boiteux S, Hickson ID, Radicella JP. XRCC1 coordinates the initial and late stages of DNA abasic site repair through protein–protein interactions. EMBO J. 2001;20:6530–9.PubMedCentralCrossRefPubMedGoogle Scholar
  11. 11.
    Sossou M, Flohr-Beckhaus C, Schulz I, Daboussi F, Epe B, Radicella JP. APE1 overexpression in XRCC1-deficient cells complements the defective repair of oxidative single strand breaks but increases genomic instability. Nucleic Acids Res. 2005;33(1):298–306.PubMedCentralCrossRefPubMedGoogle Scholar
  12. 12.
    Ashraf MJ, Maghbul M, Azarpira N, Khademi B. Expression of Ki67 and P53 in primary squamous cell carcinoma of the larynx. Indian J Pathol Microbiol. 2010;53:661–5.CrossRefPubMedGoogle Scholar
  13. 13.
    Tang QL, Fan S, Li HG, Chen WL, Shen XM, Yuan XP, et al. Expression of Cyr61 in primary salivary adenoid cystic carcinoma and its relation to Ki-67 and prognosis. Oral Oncol. 2011;47:365–70.CrossRefPubMedGoogle Scholar
  14. 14.
    Okabe M, Inagaki H, Murase T, Inoue M, Nagai N, Eimoto T. Prognostic significance of p27 and Ki-67 expression in mucoepidermoid carcinoma of the intraoral minor salivary gland. Mod Pathol. 2001;14:1008–14.CrossRefPubMedGoogle Scholar
  15. 15.
    Schmilovitz-Weiss H, Tobar A, Halpern M, Levy I, Shabtai E, Ben-Ari Z. Tissue expression of squamous cellular carcinoma antigen and Ki67 in hepatocellular carcinoma-correlation with prognosis: a historical prospective study. Diagn Pathol. 2011;6:121.PubMedCentralCrossRefPubMedGoogle Scholar
  16. 16.
    Faratian D, Munro A, Twelves C, Bartlett JM. Membranous and cytoplasmic staining of Ki67 is associated with HER2 and ER status in invasive breast carcinoma. Histopathology. 2009;54:254–7.CrossRefPubMedGoogle Scholar
  17. 17.
    Han B, Lin S, Yu LJ, Wang RZ, Wang YY. Correlation of 18 F-FDG PET activity with expressions of survivin, Ki67, and CD34 in non-small-cell lung cancer. Nucl Med Commun. 2009;30:831–7.CrossRefPubMedGoogle Scholar
  18. 18.
    Fischer CA, Jung M, Zlobec I, Green E, Storck C, Tornillo L, et al. Co-overexpression of p21 and Ki-67 in head and neck squamous cell carcinoma relative to a significantly poor prognosis. Head Neck. 2011;33:267–73.CrossRefPubMedGoogle Scholar
  19. 19.
    Mahjabeen I, Baig RM, Masood N, Sabir M, Malik FA, Kayani MA. OGG1 Gene sequence variation in head and neck cancer patients in Pakistan. Asian Pac J Cancer Prev. 2011;12:2779–83.PubMedGoogle Scholar
  20. 20.
    Mahjabeen I, Baig RM, Masood N, Sabir M, Inayat U, Malik FA, et al. Novel mutations of OGG1 base excision repair pathway gene in laryngeal cancer patients. Fam Cancer. 2012;11(4):587–93.CrossRefPubMedGoogle Scholar
  21. 21.
    Mahjabeen I, Baig RM, Sabir M, Kayani MA. Genetic and expressional variations of APEX1 are associated with increased risk of head and neck cancer. Mutagenesis. 2013;28(2):213–8.CrossRefPubMedGoogle Scholar
  22. 22.
    Mahjabeen I, Baig RM, Masood N, Sabir M, Inayat U, Malik FA, et al. Genetic variations in XRCC1 gene in sporadic head and neck cancer (HNC) patients. Pathol Oncol Res. 2012;19(20):183–8.PubMedGoogle Scholar
  23. 23.
    Mahjabeen I, Chen Z, Zhou X, Kayani MA. Decreased mRNA expression levels of base excision repair (BER) pathway genes is associated with enhanced Ki-67 expression in HNSCC. Med Oncol. 2012;29(5):3620–5.CrossRefPubMedGoogle Scholar
  24. 24.
    Wang YX, Sun YE, Li XH, Wang ZB, Tong XY, Liu YL. Comparative study on molecular staging of lymph nodes in non-small cell lung cancer patients. Ai Zheng. 2009;28(3):318–22.PubMedGoogle Scholar
  25. 25.
    Al-Moundhri MS, Nirmala V, Al-Hadabi I, Al-Mawaly K, Burney I, Al-Nabhani M, et al. The prognostic significance of p53, p27 kip1, p21 waf1, HER-2/neu, and Ki67 proteins expression in gastric cancer: a clinicopathological and immunohistochemical study of 121 Arab patients. J Surg Oncol. 2005;91(4):243–52.CrossRefPubMedGoogle Scholar
  26. 26.
    Mambo E, Chatterjee A, Souza-Pinto NC, Mayard S, Hogue BA, Hoque M, et al. Oxidized guanine lesions and hOgg1 activity in lung cancer. Oncogene. 2005;24:4496–508.CrossRefPubMedGoogle Scholar
  27. 27.
    Kunisada M, Sakumi K, Tominaga Y, Budiyanto A, Ueda M, Ichihashi M, et al. 8-Oxoguanine formation induced by chronic UVB exposure makes Ogg1 knockout mice susceptible to skin carcinogenesis. Cancer Res. 2005;65:6006–10.CrossRefPubMedGoogle Scholar
  28. 28.
    Huang XX, Scolyer RA, Abubakar A, Halliday GM. Human 8-oxoguanine-DNA glycosylase-1 is downregulated in human basal cell carcinoma. Mol Genet Metab. 2012;106:127–30.CrossRefPubMedGoogle Scholar
  29. 29.
    Karihtala P, Kauppila S, Puistola U, Jukkola-Vuorinen A. Absence of the DNA repair enzyme human 8-oxoguanine glycosylase is associated with an aggressive breast cancer phenotype. Br J Cancer. 2012;106:344–7.PubMedCentralCrossRefPubMedGoogle Scholar
  30. 30.
    Salim EI, Morimura K, Menesi A, El-Lity M, Fukushima S, Wanibuchi H. Elevated oxidative stress and DNA damage and repair levels in urinary bladder carcinomas associated with schistosomiasis. Int J Cancer. 2008;123:601–8.CrossRefPubMedGoogle Scholar
  31. 31.
    Ku YP, Jin M, Kim KH, Ahn YJ, Yoon SP, You HJ, et al. Immunolocalization of 8-OHdG and OGG1 in pancreatic islets of streptozotocin-induced diabetic rats. Acta Histochem. 2009;111:138–44.CrossRefPubMedGoogle Scholar
  32. 32.
    Preston TJ, Henderson JT, McCallum GP, Wells PG. Base excision repair of reactive oxygen species-initiated 7, 8- dihydro-8-oxo-2′-deoxyguanosine inhibits the cytotoxicity of platinum anticancer drugs. Mol Cancer Ther. 2009;8(7):2015–26.PubMedCentralCrossRefPubMedGoogle Scholar
  33. 33.
    Moore DH, Michael H, Tritt R, Parsons SH, Kelley MR. Alternations in the expression of the DNA repair/ redox enzyme APE/ref-1 in epithelial ovarian cancers. Clin Cancer Res. 2000;6:602–9.PubMedGoogle Scholar
  34. 34.
    Kelley MR, Cheng L, Foster R, Tritt R, Jiang J, Broshears J, et al. Elevated and altered expression of the multifunctional DNA base excision repair and redox enzyme Ape1/ref-1 in prostate cancer. Clin Cancer Res. 2001;7:824–30.PubMedGoogle Scholar
  35. 35.
    Bobola MS, Finn LS, Ellenbogen RG, Geyer JR, Berger MS, Braga JM, et al. Apurinic/apyrimidinic endonuclease activity is associatedwith response to radiation and chemotherapy in medulloblastoma and primitive neuroectodermal tumors. Clin Cancer Res. 2005;11(20):7405–14.CrossRefPubMedGoogle Scholar
  36. 36.
    Yang S, Irani K, Heffron SE, Jurnak F, Meyskens FLJ. Alterations in the expression of the apurinic/apyrimidinic endonuclease-1/redox factor-1 (APE/ref-1) in human melanoma and identification of the therapeutic potential of resveratrol as an APE/ref-1 inhibitor. Mol Cancer Ther. 2005;4(12):1923–35.CrossRefPubMedGoogle Scholar
  37. 37.
    Raffoul JJ, Banerjee S, Singh-Gupta V, Knoll ZV, Fite A, Zhang H, et al. Down-regulation of apurinic/apyrimidinic endonuclease 1/redox factor-1 expression by soy isoflavones enhances prostate cancer radiotherapy in vitro and in vivo. Cancer Res. 2007;67(5):2141–9.CrossRefPubMedGoogle Scholar
  38. 38.
    Jiang L, Liu X, Chen Z, Jin Y, Heidbreder CE, Kolokythas A, et al. MicroRNA-7 targets insulin-like growth factor 1 receptor (IGF1R) in tongue squamous cell carcinoma cells. Biochem J. 2010;432:199–205.PubMedCentralCrossRefPubMedGoogle Scholar
  39. 39.
    Fishel ML, Jiang Y, Rajeshkumar NV, Scandura G, Sinn AL, He Y, et al. Impact of APE1/ref-1 redox inhibition on pancreatic tumor growth. Mol Cancer Ther. 2011;10(9):1698–708.PubMedCentralCrossRefPubMedGoogle Scholar
  40. 40.
    Schindl M, Oberhuber G, Pichlbauer EG, Obermair A, Birner P, Kelley MR. DNA repair-redox enzyme apurinic endonuclease in cervical cancer: evaluation of redox control of HIF-1α and prognostic significance. Int J Oncol. 2001;19:799–802.PubMedGoogle Scholar
  41. 41.
    Freitas S, Moore DH, Michael H, Kelley MR. Studies of apurinic/apyrimidinic endonuclease/ref-1 expression in epithelial ovarian cancer: correlations with tumor progression and platinum resistance. Clin Cancer Res. 2003;9:4689–94.PubMedGoogle Scholar
  42. 42.
    Lee JW, Jin J, Rha KS, Kim YM. Expression pattern of apurinic/apyrimidinic endonuclease in sinonasal squamous cell carcinoma. Otolaryngol Head Neck Surg. 2012;147(4):788–95.CrossRefPubMedGoogle Scholar
  43. 43.
    Ford BN, Ruttan CC, Kyle VL, Brackley ME, Glickman BW. Identification of single nucleotide polymorphisms in human DNA repair genes. Carcinogenesis. 2000;21:1977–81.CrossRefPubMedGoogle Scholar
  44. 44.
    Hu JJ, Smith TR, Miller MS, Mohrenweiser HW, Golden A, Case LD. Amino acid substitution variants of APE1 and XRCC1 genes associated with ionizing radiation sensitivity. Carcinogenesis. 2001;22:917–22.CrossRefPubMedGoogle Scholar
  45. 45.
    Liu W, AO L, Cui Z, Zhou Z, Zhou Y, Yuan X, et al. Molecular analysis of DNA repair gene methylation and protein expression during chemical-induced rat lung carcinogenesis. Biochem Biophys Res Commun. 2011;408:595–601.CrossRefPubMedGoogle Scholar
  46. 46.
    Wang P, Tang JT, Peng YS, Chen XY, Zhang YJ, Fang JY. XRCC1 downregulated through promoter hypermethylation is involved in human gastric carcinogenesis. J Dig Dis. 2010;11:343–51.CrossRefPubMedGoogle Scholar
  47. 47.
    Wang S, Wu X, Chen Y, Zhang J, Ding J, Zhou Y, et al. Prognostic and predictive role of JWA and XRCC1 expressions in gastric cancer. Clin Cancer Res. 2012;18(10):2987–96.CrossRefPubMedGoogle Scholar
  48. 48.
    Chetty C, Dontula R, Gujrati M, Dinh DH, Lakka SS. Blockade of SOX4 mediated DNA repair by SPARC enhances radioresponse in medulloblastoma. Cancer Lett. 2012;323:188–98.PubMedCentralCrossRefPubMedGoogle Scholar
  49. 49.
    Sak SC, Harnden P, Johnston CF, Paul AB, Kiltie AE. APE1 and XRCC1 protein expression levels predict cancer-specific survival following radical radiotherapy in bladder cancer. Clin Cancer Res. 2005;11:6205.CrossRefPubMedGoogle Scholar
  50. 50.
    Cheng X, Lu W, Ye F, Wan X, Xie X. The association of XRCC1 gene single nucleotide polymorphisms with response to neoadjuvant chemotherapy in locally advancedcervical carcinoma. J Exp Clin Cancer Res. 2009;28:91.PubMedCentralCrossRefPubMedGoogle Scholar
  51. 51.
    Kang CH, Jang BG, Kim DW, Chung DH, Kim YT, Jheon S, et al. The prognostic significance of ERCC1, BRCA1, XRCC1, and betaIII-tubulin expression in patients with non-small cell lung cancer treated by platinum- and taxane-based neoadjuvant chemotherapy and surgical resection. Lung Cancer. 2010;68:478–83.CrossRefPubMedGoogle Scholar
  52. 52.
    Ang MK, Patel MR, Yin XY, Sundaram S, Fritchie K, Zhao N, et al. High XRCC1 protein expression is associated with poorer survival in patients with head and neck squamous cell carcinoma. Clin Cancer Res. 2011;17(20):6542–52.PubMedCentralCrossRefPubMedGoogle Scholar
  53. 53.
    Rybarova S, Vecanova J, Hodorova I, Mihalik J, Cizmarikova M, Mojzis J, et al. Association between polymorphisms of XRCC1, p53 and MDR1 genes, the expression of their protein products and prognostic significance in human breast cancer. Med Sci Monit. 2011;17:BR354–63.PubMedCentralCrossRefPubMedGoogle Scholar
  54. 54.
    Fujimura M, Morita-Fujimura Y, Noshita N, Yoshimoto T, Chan PH. Reduction of the DNA base excision repair protein, XRCC1, may contribute to DNA fragmentation after cold injury-induced brain trauma in mice. Brain Res. 2000;869:105–11.CrossRefPubMedGoogle Scholar
  55. 55.
    Rodrigues RB, Motta Rda R, Machado SM, Cambruzzi E, Zettler EW, Zettler CG, et al. Prognostic value of the immunohistochemistry correlation of Ki-67 and p53 in squamous cell carcinomas of the larynx. Braz J Otorhinolaryngol. 2008;74:855–9.PubMedGoogle Scholar
  56. 56.
    Kim SJ, Shin HJ, Jung K, Baek S, Shin BK, Choi J, et al. Prognostic value of carbonic anhydrase IX and Ki-67 expression in squamous cell carcinoma of the tongue. Jpn J Clin Oncol. 2007;37:812–9.CrossRefPubMedGoogle Scholar
  57. 57.
    Szczuraszek K, Mazur G, Jelen M, Dziegiel P, Surowiak P, Zabel M. Prognostic significance of Ki-67 antigen expression in non-Hodgkin’s lymphomas. Anticancer Res. 2008;28:1113–8.PubMedGoogle Scholar
  58. 58.
    Cheang MCU, Chia SK, Voduc D, Gao D, Leung S, Snider J, et al. Ki67 index, HER2 status, and prognosis of patients with luminal B breast cancer. JNCI. 2009;101(10):736–50.PubMedCentralCrossRefPubMedGoogle Scholar
  59. 59.
    Sym S, Hong J, Cho E, Lee W, Chung M, Ha S, Park Y, Park J, Lee J, Shin D (2011) Prognostic impact of immunohistochemical expression of Ki-67 in patients with advanced gastric cancer who underwent curative Resection. J Clin Oncol 29Google Scholar
  60. 60.
    Jiang Y, Cheng B, Ge M, Zhang G. The prognostic significance of p63 and Ki-67 expression in myoepithelial carcinoma. Head Neck Oncol. 2012;4:9.PubMedCentralCrossRefPubMedGoogle Scholar
  61. 61.
    Habib SL. Insight into mechanism of oxidative DNA damage in angiomyolipomas from TSC patients. Mol Cancer. 2009;8:13.PubMedCentralCrossRefPubMedGoogle Scholar
  62. 62.
    Jiang Y, Zhou S, Sandusky GE, Kelley MR, Fishel ML. Reduced expression of DNA repair and redox signaling protein APE1/ref-1 impairs human pancreatic cancer cell survival, proliferation, and cell cycle progression. Cancer Investig. 2010;228:885–95.CrossRefGoogle Scholar
  63. 63.
    Al-Attar A, Gossage L, Fareed KR, Shehata M, Mohammed M, Zaitoun AM, et al. Human apurinic/apyrimidinic endonuclease (APE1) is a prognostic factor in ovarian, gastro-oesophageal and pancreatico-biliary cancers. Br J Cancer. 2010;102:704–9.PubMedCentralCrossRefPubMedGoogle Scholar
  64. 64.
    Grosch S, Kaina B. Transcriptional activation of apurinic/apyrimidinic endonuclease (Ape, ref-1) by oxidative stress requires CREB. Biochem Biophys Res Commun. 1999;261:859–63.CrossRefPubMedGoogle Scholar
  65. 65.
    Vaezi A, Feldman CH, Niedernhofer LJ. ERCC1 and XRCC1 as biomarkers for lung and head and neck cancer. Pharmgenomics Pers Med. 2011;4:47–63.PubMedCentralPubMedGoogle Scholar
  66. 66.
    Krskova L, Kalinova M, Brızova H, Mrhalova M, Sumerauer D, Kodet R. Molecular and immunohistochemical analyses of BCL2, KI-67, and cyclin D1 expression in synovial sarcoma. Cancer Genet Cytogenet. 2009;193:1–8.CrossRefPubMedGoogle Scholar
  67. 67.
    Brouwer-Visser J, Cossio MJ, Chao SK, Huang GS. Effect of IGF2 overexpression on the tumorigenicity of human ovarian carcinoma cells. Cancer Res. 2012;72:1.CrossRefGoogle Scholar
  68. 68.
    Ma J, Li J, Li H, Xiao X, Shen L, Fang L. Downregulation of pancreatic-duodenal homeobox 1 expression in breast cancer patients: a mechanism of proliferation and apoptosis in cancer. Mol Med Rep. 2012;6:983–8.PubMedGoogle Scholar
  69. 69.
    Urruticoechea A, Smith IE, Dowsett M. Proliferation marker Ki-67 in early breast cancer. J Clin Oncol. 2005;23:7212–20.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Ishrat Mahjabeen
    • 1
    • 2
  • Kashif Ali
    • 1
  • Xiaofeng Zhou
    • 2
    • 3
  • Mahmood Akhtar Kayani
    • 1
  1. 1.Cancer Genetics Lab, Department of BiosciencesCOMSATS Institute of Information and TechnologyIslamabadPakistan
  2. 2.Center for Molecular Biology of Oral Diseases, College of DentistryUniversity of Illinois at ChicagoChicagoUSA
  3. 3.Department of Periodontics, College of Dentistry, Graduate College, UIC Cancer CenterUniversity of Illinois at ChicagoChicagoUSA

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