Expression of Endogenous Hypoxia Markers in Vulvar Squamous Cell Carcinoma

  • Li, Yu-Zhu (Department of Integrated Traditional Chinese and Western Medicine, Qilu Hospital, Shandong University) ;
  • Li, Shu-Ling (Department of Integrated Traditional Chinese and Western Medicine, Qilu Hospital, Shandong University) ;
  • Li, Xia (Institute of Basic Medicine, Shandong Academy of Medical Science) ;
  • Wang, Li-Jie (Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University) ;
  • Wang, Jiu-Ling (Department of Integrated Traditional Chinese and Western Medicine, Qilu Hospital, Shandong University) ;
  • Xu, Jia-Wen (Department of Pathology, Provincial Hospital Affiliated to Shandong University) ;
  • Wu, Zhi-Hong (Department of Integrated Traditional Chinese and Western Medicine, Qilu Hospital, Shandong University) ;
  • Gong, Li (Department of Integrated Traditional Chinese and Western Medicine, Qilu Hospital, Shandong University) ;
  • Zhang, Xiao-Dan (Department of Integrated Traditional Chinese and Western Medicine, Qilu Hospital, Shandong University)
  • Published : 2012.08.31


Objective: To investigate the expression of endogenous hypoxia-related markers identified as being involved in vulvar squamous cell carcinoma (VSCC). Methods: We performed immunohistochemical staining of hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$), glucose transporter-1 (GLUT-1), carbonic anhydrase 9 (CA-9) and vascular endothelial growth factor (VEGF), on tissue sections of 25 VSCC patients, 10 vulvar intraepithelial neoplasia (VIN) patients and 12 healthy controls. Results: HIF-$1{\alpha}$ expression was found in all sections, with no significant difference between controls, VIN and VSCC sections (all P<0.05). Glut-1 expression was found in 25% of control, 90% of VIN and 100% of VSCC sections. A significant difference between control and VIN or VSCC was observed (all P<0.05), while no difference was found between VIN and VSCC sections (P>0.05). CA-9 expression was negative in control sections, but it was found in 30% of VIN sections and 52% of VSCC sections with strong staining. Similarly, CA-9 expression also showed obvious differences between controls and VIN or VSCC sections (all P<0.05). However, there was no significant difference between VIN and VSCC (P>0.05). There were only 25% of control sections with weak VEGF expression, while strong staining was found in about 60% of VIN sections and 25% of VSCC sections (all P<0.05). In addition, a difference was also found between VIN and VSCC sections (P<0.05). Conclusion: Expression of endogenous hypoxia markers (HIF-$1{\alpha}$, GLUT-1, CA-9 and VEGF) might be involved in the malignant progression of VSCC.


  1. Airley RE, Loncaster J, Raleigh JA, et al (2003). GLUT-1 and CAIX as intrinsic markers of hypoxia in carcinoma of the cervix: relationship to pimonidazole binding. Int J Cancer, 104, 85-91.
  2. Birner P, Schindl M, Obermair A, et al (2000). Overexpression of hypoxia-inducible factor 1alpha is a marker for an unfavorable prognosis in early-stage invasive cervical cancer. Cancer Res, 60, 4693-6.
  3. Brown JM (2000). Exploiting the hypoxic cancer cell: mechanisms and therapeutic strategies. Mol Med Today, 6, 157-62.
  4. Bussink J, Kaanders JH, van der Kogel AJ (2003). Tumor hypoxia at the micro-regional level: clinical relevance and predictive value of exogenous and endogenous hypoxic cell markers. Radiother Oncol, 67, 3-15.
  5. Carmeliet P, Jain RK (2000). Angiogenesis in cancer and other diseases. Nature, 407, 249-57.
  6. Choi SW, Kim JY, Park JY, et al (2008). Expression of carbonic anhydrase IX is associated with postoperative recurrence and poor prognosis in surgically treated oral squamous cell carcinoma. Hum Pathol, 39, 1317-22.
  7. Coleman CN, Mitchell JB, Camphausen K (2002). Tumor hypoxia: chicken, egg, or a piece of the farm? J Clin Oncol, 20, 610-5.
  8. Danielsen T, Rofstad EK (2000). The constitutive level of vascular endothelial growth factor (VEGF) is more important than hypoxia-induced VEGF up-regulation in the angiogenesis of human melanoma xenografts. Br J Cancer, 82, 1528-34.
  9. Davda S, Bezabeh T (2006). Advances in methods for assessing tumor hypoxia in vivo: implications for treatment planning. Cancer Metastasis Rev, 25, 469-80.
  10. Davies CA, Jeziorska M, Freemont AJ, et al (2006). The differential expression of VEGF, VEGFR-2, and GLUT-1 proteins in disease subtypes of systemic sclerosis. Hum Pathol, 37, 190-7.
  11. Fang JS, Gillies RD, Gatenby RA (2008). Adaptation to hypoxia and acidosis in carcinogenesis and tumor progression. Semin Cancer Biol, 18, 330-7.
  12. Ferrara N (2002). VEGF and the quest for tumour angiogenesis factors. Nat Rev Cancer, 2, 795-803.
  13. Gillies RJ, Gatenby RA. (2007). Hypoxia and adaptive landscapes in the evolution of carcinogenesis. Cancer Metastasis Rev, 26, 311-7.
  14. Harris AL (2002). Hypoxia--a key regulatory factor in tumour growth. Nat Rev Cancer, 2, 38-47.
  15. Hendriksen EM, Span PN, Schuuring J, et al (2009). Angiogenesis, hypoxia and VEGF expression during tumour growth in a human xenograft tumour model. Microvasc Res, 77, 96-103.
  16. Hua Z, Lv Q, Ye W, et al (2006). MiRNA-directed regulation of VEGF and other angiogenic factors under hypoxia. PLoS One, 1, e116.
  17. Iida T, Yasuda M, Miyazawa M, et al (2008). Hypoxic status in ovarian serous and mucinous tumors: relationship between histological characteristics and HIF-1alpha/GLUT-1 expression. Arch Gynecol Obstet, 277, 539-46.
  18. Jiang BH, Semenza GL, Bauer C, et al (1996). Hypoxia-inducible factor 1 levels vary exponentially over a physiologically relevant range of O2 tension. Am J Physiol, 271, C1172-80.
  19. Jonathan RA, Wijffels KI, Peeters W, et al (2006). The prognostic value of endogenous hypoxia-related markers for head and neck squamous cell carcinomas treated with ARCON. Radiother Oncol, 79, 288-97.
  20. Jubb AM, Pham TQ, Hanby AM, et al (2004). Expression of vascular endothelial growth factor, hypoxia inducible factor 1alpha, and carbonic anhydrase IX in human tumours. J Clin Pathol, 57, 504-12.
  21. Kaluz S, Kaluzova M, Liao SY, et al (2009). Transcriptional control of the tumor- and hypoxia-marker carbonic anhydrase 9: A one transcription factor (HIF-1) show? Biochim Biophys Acta, 1795, 162-72.
  22. Krieg M, Haas R, Brauch H, et al (2000). Up-regulation of hypoxia-inducible factors HIF-1alpha and HIF-2alpha under normoxic conditions in renal carcinoma cells by von Hippel- Lindau tumor suppressor gene loss of function. Oncogene, 19, 5435-43.
  23. Le QT, Kong C, Lavori PW, et al (2007). Expression and prognostic significance of a panel of tissue hypoxia markers in head-and-neck squamous cell carcinomas. Int J Radiat Oncol Biol Phys, 69, 167-75.
  24. Liao D, and Johnson RS (2007). Hypoxia: a key regulator of angiogenesis in cancer. Cancer Metastasis Rev, 26, 281-90.
  25. Linden T, Katschinski DM, Eckhardt K, et al (2003). The antimycotic ciclopirox olamine induces HIF-1alpha stability, VEGF expression, and angiogenesis. FASEB J, 17, 761-3.
  26. Loncaster JA, Harris AL, Davidson SE, et al (2001). Carbonic anhydrase (CA IX) expression, a potential new intrinsic marker of hypoxia: correlations with tumor oxygen measurements and prognosis in locally advanced carcinoma of the cervix. Cancer Res, 61, 6394-9.
  27. Mizukami Y, Kohgo Y, Chung DC (2007). Hypoxia inducible factor-1 independent pathways in tumor angiogenesis. Clin Cancer Res, 13, 5670-4.
  28. Semenza GL (2003). Targeting HIF-1 for cancer therapy. Nat Rev Cancer, 3, 721-32.
  29. Ortega AD, Sanchez-Arago M, Giner-Sanchez D, et al (2009). Glucose avidity of carcinomas. Cancer Lett, 276, 125-35.
  30. Osinsky S, Zavelevich M, Vaupel P (2009). Tumor hypoxia and malignant progression. Exp Oncol, 31, 80-6.
  31. Pecorelli S (2009). Revised FIGO staging for carcinoma of the vulva, cervix, and endometrium. Int J Gynaecol Obstet, 105, 103-4.
  32. Rademakers SE, Lok J, van der Kogel AJ, et al (2011). Metabolic markers in relation to hypoxia; staining patterns and colocalization of pimonidazole, HIF-1alpha, CAIX, LDH-5, GLUT-1, MCT1 and MCT4. BMC Cancer, 11, 167.
  33. Saidi A, Javerzat S, Bellahcene A, et al (2008). Experimental anti-angiogenesis causes upregulation of genes associated with poor survival in glioblastoma. Int J Cancer, 122, 2187-98.
  34. Seeber LM, Horree N, Vooijs MA, et al (2011). The role of hypoxia inducible factor-1alpha in gynecological cancer. Crit Rev Oncol Hematol, 78, 173-84.
  35. Semenza GL (2009). Regulation of cancer cell metabolism by hypoxia-inducible factor 1. Semin Cancer Biol, 19, 12-6.
  36. Semenza GL (2010). HIF-1: upstream and downstream of cancer metabolism. Curr Opin Genet Dev, 20, 51-6.
  37. Semenza GL (2012). Hypoxia-inducible factors: mediators of cancer progression and targets for cancer therapy. Trends Pharmacol Sci, 33, 207-14.
  38. Shibuya M. (2001). Structure and function of VEGF/VEGFreceptor system involved in angiogenesis. Cell Struct Funct, 26, 25-35.
  39. Talks KL, Turley H, Gatter KC, et al (2000). The expression and distribution of the hypoxia-inducible factors HIF-1alpha and HIF-2alpha in normal human tissues, cancers, and tumorassociated macrophages. Am J Pathol, 157, 411-21.
  40. Tian M, Zhang H, Nakasone Y, et al (2004). Expression of Glut- 1 and Glut-3 in untreated oral squamous cell carcinoma compared with FDG accumulation in a PET study. Eur J Nucl Med Mol Imaging, 31, 5-12.
  41. Tyring SK (2003). Vulvar squamous cell carcinoma: guidelines for early diagnosis and treatment. Am J Obstet Gynecol, 189, S17-23.
  42. van de Nieuwenhof HP, Bulten J, Hollema H, et al (2011). Differentiated vulvar intraepithelial neoplasia is often found in lesions, previously diagnosed as lichen sclerosus, which have progressed to vulvar squamous cell carcinoma. Mod Pathol, 24, 297-305.
  43. van de Nieuwenhof HP, Massuger LF, van der Avoort IA, et al (2009). Vulvar squamous cell carcinoma development after diagnosis of VIN increases with age. Eur J Cancer, 45, 851-6.
  44. van den Beucken T, Koritzinsky M, Niessen H, et al (2009). Hypoxia-induced expression of carbonic anhydrase 9 is dependent on the unfolded protein response. J Biol Chem, 284, 24204-12.
  45. Vordermark D, Brown JM (2003). Endogenous markers of tumor hypoxia predictors of clinical radiation resistance? Strahlenther Onkol, 179, 801-11.
  46. Wenger RH (2002). Cellular adaptation to hypoxia: O2-sensing protein hydroxylases, hypoxia-inducible transcription factors, and O2-regulated gene expression. FASEB J, 16, 1151-62.
  47. Yasuda M, Miyazawa M, Fujita M, et al (2008). Expression of hypoxia inducible factor-1alpha (HIF-1alpha) and glucose transporter-1 (GLUT-1) in ovarian adenocarcinomas: difference in hypoxic status depending on histological character. Oncol Rep, 19, 111-6.
  48. Zhong H, De Marzo AM, Laughner E, et al (1999). Overexpression of hypoxia-inducible factor 1alpha in common human cancers and their metastases. Cancer Res, 59, 5830-5.

Cited by

  1. Maligne Tumore der Vulva: eine Übersicht für den Dermatologen vol.165, pp.7-8, 2015,