Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Microvessel density is high in clear-cell renal cell carcinomas of Ukrainian patients exposed to chronic persistent low-dose ionizing radiation after the Chernobyl accident


During the 25-year period subsequent to the Chernobyl accident, the morbidity of malignant renal tumors in Ukraine has increased from 4.7 to 10.7 per 100,000 of the total population. Recent studies of our group have shown that increases in morbidity, aggressiveness, and proliferative activity of renal cell carcinomas (RCCs), especially clear-cell renal cell carcinoma (CCRCC), in Ukrainian patients continuously inhabiting the radio-contaminated areas could be explained by specific molecular changes influenced by the so-called “chronic persistent low-dose ionizing radiation” (CPLDIR) exposure. This study aimed to examine the role of angiogenesis in CCRCC carcinogenesis associated with CPLDIR in patients living more than 20 years in cesium 137 (137Cs) contaminated areas after the Chernobyl accident in Ukraine. Paraffin-embedded specimens of 106 CCRCs were studied: Control cases were 18 tumors from Spanish patients (group 1), 25 tumors from Ukrainian patients from so-called clean areas without known radio-contamination (group 2), and 63 tumors from Ukrainian patients from radio-contaminated areas (group 3). For intratumoral microvessel density (MVD) determination, anti-CD31 antibody was used. A computerized image analysis program was used to quantitatively calculate the vascular density. Seventy-three percent of group 3 and 72 % of group 2 CCRCCs displayed the highest MVD. A striking increase in MVD was seen in group 3 CCRCCs, in comparison with groups 1 and 2 (p < 0.001). The majority of the hot spot vessels in group 3 was poorly differentiated. Moreover, MVD values for total vessels as well as for capillaries and tumor grade were strongly correlated. When we compared only tumor–node–metastasis tumor stages I and II, the differences remained statistically significant (p < 0.1). The ratio of the average total vessels and capillaries in the Ukrainian groups combined was 1.65:1 in comparison to the Spanish group. Our results provide evidence that CPLDIR exposure increases MVD (particularly capillary) in CCRCCs and is associated with a higher histological grade.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. 1.

    Yagoda A, Petrylak D, Thompson S (1993) Cytotoxic chemotherapy for advanced renal cell carcinoma. Urol Clin Nort Am 20:303–321

  2. 2.

    Fossa SD (2000) Interferon in metastatic renal cell carcinoma. Semin Oncol 27:187–193

  3. 3.

    Dennis Smith A, Lieber ML, Shah SN (2010) Assessing tumor response and detecting recurrence in metastatic renal cell carcinoma on targeted therapy: importance of size and attenuation on contrast enhanced CT. Am J Roentgenol 194:157–165

  4. 4.

    Saydackova NA, Starceva LM, Kravchuk NC (2007) The state of urological assistance for the population in Ukraine. Annual report. Ministry of Health in Ukraine, Kiev, pp 146–153

  5. 5.

    Romanenko A, Morrell-Quadreny L, Nepomnyaschy V, Vozianov A, Llombart-Bosch A (2000) Pathology and proliferative activity of renal-cell carcinomas and renal oncocytomas in patients with different radiation exposure after the Chernobyl accident in Ukraine. Int J Cancer 87:880–883

  6. 6.

    Romanenko A, Morrell-Quadreny L, Nepomnyaschy V, Vozianov A, Llombart-Bosch A (2001) Radiation sclerosing proliferative atypical nephropathy of peritumoral tissue of renal-cell carcinomas after the Chernobyl accident in Ukraine. Virchows Arch 438:146–153

  7. 7.

    Romanenko A, Morrell-Quadreny L, Lopez-Guerrero JA, Pellin A, Nepomnyaschy V, Vozianov A, Llombart-Bosch A (2004) The INK4a/ARF locus: role in cell cycle control for renal cell epithelial tumor growth after the Chernobyl accident. Virchows Arch 445:298–304

  8. 8.

    Romanenko A, Morrell-Quadreny L, Ramos D, Vozianov A, Llombart-Bosch A (2006) Alteration of apoptotic regulatory molecules in conventional renal cell carcinoma influenced by chronic long-term low-dose radiation exposure in humans revealed by tissue microarray. Cancer Genomics Proteomics 3:107–112

  9. 9.

    Romanenko A, Morrell-Quadreny L, Ramos D, Nepomnyaschy V, Vozianov A, Llombart-Bosch A (2006) Extracellular matrix alterations in conventional renal cell carcinomas by tissue microarray profiling influenced by persistent, long-term, low-dose ionizing radiation exposure in humans. Virchows Arch 448:584–590

  10. 10.

    Clarke RH (2001) Control of low-level radiation exposure: what is the problem and how can it be solved? Health Phys 80:391–396

  11. 11.

    Christensen R, Alsner J, Brandt Sorensen F, Dagnaes-Hansen F, Kolvraa S, Serakinci N (2008) Transformation of human mesenchymal stem cells in radiation carcinogenesis: long-term effect of ionizing tadiation. Regen Med 3(6):849–861

  12. 12.

    Mullenders L, Atkinson M, Peretzke H, Sabatier L, Bouffler S (2009) Assessing cancer risks of low-dose radiation. Nat Rev Cancer 9(8):596–604

  13. 13.

    Wilson PF, Nham PB, Urbin SS, Hinz JM, Jones IM, Thompson LH (2010) Inter-individual variation in DNA double-strand break repair in human fibroblasts before and after exposure to low doses of ionizing radiation. Mutat Res 683:91–97

  14. 14.

    Bergers G, Benjamin LE (2003) Tumorigenesis and the angiogenic switch. Nat Rev Cancer 3:401–410

  15. 15.

    Kerbel RS (2008) Tumor angiogenesis. N Engl J Med 358:2039–2049

  16. 16.

    Zhang Y, Jiang X, Qin X, Ye D, Yi Z, Liu M, Bai O, Fang J, Chen Y (2010) RKTG inhibits angiogenesis by suppressing MAPK-mediated autocrine VEGF signaling and is downregulated in clear-cell renal cell carcinoma. Oncogene 29:5404–5415

  17. 17.

    Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144:646–674

  18. 18.

    Ravasio R, Ortega C, Sabbatini R, Porta C (2011) Bevacizumab plus interferon-a versus sunititab for first-line treatment of renal cell carcinoma in Italy: a cost-minimization analysis. Clin Drug Investig 31:507–517

  19. 19.

    North S, Moenner M, Bikfalvi A (2005) Recent developments in the regulation of the “angiogenic switch” by cellular stress factors in tumors. Cancer Lett 218:1–14

  20. 20.

    Ushio-Fukai M, Nakamura Y (2008) Reactive oxygen species and angiogenesis: NADPH oxidase as target for cancer therapy. Cancer Lett 266:37–52

  21. 21.

    Kim J, Koyanagi T, Mochly-Rosen D (2011) PKCo activation mediates angiogenesis via NADPH oxidase activity in PC-3 prostate cancer cells. Prostate 71:946–954

  22. 22.

    Weidner N (1995) Current pathologic methods for measuring intratumoral microvessel density within breast carcinoma and other solid tumors. Breast Cancer Res Treat 36:169–180

  23. 23.

    Uzzan B, Nicolas P, Cucherat M, Perret GY (2004) Microvessel density as a prognostic factor in women with breast cancer: a systematic review of the literature and meta-analysis. Cancer Res 64:2941–2955

  24. 24.

    Delahunt B, Bethwaite PB, Thornton A (1997) Prognostic significance of microscopic vascularity for clear cell renal cell carcinoma. Br J Urol 80:401–404

  25. 25.

    Herbst C, Kosmehl H, Stiller KJ (1998) Evaluation of microvessel density by computerized image analysis in human renal cell carcinoma. Correlation to pT category, nuclear grade, proliferative activity and occurrence of metastasis. J Cancer Res Clin Oncol 124:141–147

  26. 26.

    Gontero P, Ceratti G, Guglielmetti S, Andorno A, Terrone C, Bonvini D, Faggiano F, Tizzani A, Frea B, Valente G (2008) Prognostic factors in a prospective series of papillary renal cell carcinoma. BJU Int 102:697–702

  27. 27.

    Nativ O, Sabo E, Reiss A (1998) Clinical significance of tumor angiogenesis in patients with localized renal cell carcinoma. Urol 51:693–696

  28. 28.

    Yildiz E, Ayan S, Goze F, Gultekin EY (2008) Relation of microvessel density with microvascular invasion, metastasis and prognosis in renal cell carcinoma. BJU Int 101:758–754

  29. 29.

    Sharma SG, Aggarwal N, Gupta SD, Singh MK, Gupta R, Dinda AK (2010) Angiogenesis in renal cell carcinoma: correlation of microvessel density and microvessel area with other prognostic factors. Int Urol Nephrol 10:9779–9787

  30. 30.

    Yilmazer D, Han U, Onal B (2007) A comparison of the vascular density of VEGF expression with microvascular density determined with CD34 and CD31 staining and conventional prognostic markers in renal cell carcinoma. Int Urol Nephrol 39:691–698

  31. 31.

    Bhattacharyya NK, Chatterjee U, Sarkar S, Kundu AK (2008) A study of proliferative activity, angiogenesis and nuclear grading in renal cell carcinoma. Indian J Pathol Microbiol 51:17–21

  32. 32.

    Llombart-Bosch A, Lopez-Guerrero JA, Carda Batalla C, Ruiz-Sauri A, Peydro-Olaya A (2003) Structural basis of tumoral angiogenesis. Adv Exp Med Biol 532:69–89

  33. 33.

    Sobin LH and Wittekind C (eds) (2002) TNM classification of malignant tumors, 6th edn. Wiley-Liss, New York

  34. 34.

    Eble JN, Sauter G, Epstein JI, Sesterhenn IA (eds) (2004) Pathology and genetic of tumors of the urinary system and male genetic organs. World Health Organization Classification of Tumors. IARC, Lyon

  35. 35.

    Fuhrman SA, Lesky LC, Limas C (1982) Prognosis significance of morphologic parameters in renal cell carcinoma. Am J Surg Pathol 6:655–663

  36. 36.

    Sales A, Ruiz A, Llombart-Bosch A (1999) Comparative morphometric evaluation of microvessel density and nuclear area in ductal carcinoma in situ and hyperplasticductal breast lesions. Breast 8:21–25

  37. 37.

    Raes F, De Cort M, Graziani G (1991) Multi‐fractal nature of radioactivity deposition on soil after the Chernobyl accident. Health Phys 61:271–28

  38. 38.

    Weidner N, Folkman J, Pozza F, Bevilacqua P, Allred EN, Moore DH, Meli S, Gasparini G (1992) Tumor angiogenesis: a new significant and independent prognostic indicator in early-stage breast carcinoma. J Natl Cancer Inst 84:1875–1887

  39. 39.

    Lao X, Qian CN, Zhand ZF, Tan MH, Kort XJ, Resau JH, The BT (2007) Two distinct types of blood vessels in clear cell renal cell carcinoma have contrasting prognostic implications. Clin Cancer Res 13:161–169

  40. 40.

    Qian CN, Huang D, Wondergem B, The BT (2009) Complexity of tumor vasculature in clear cell renal cell carcinoma. Cancer 115:2282–2289

  41. 41.

    Fox SB (1997) Tumor angiogenesis and prognosis. Histopathol 30:294–301

  42. 42.

    Jain L, Vargo CA, Danesi R, Sissung TM, Price DK, Venzon D, Venitz J, Figg WG (2009) The role of vascular endothelial growth factor SNPs as predictive and prognostic markers for major solid tumors. Mol Cancer Ther 8:2496–2508

  43. 43.

    Shinkaruk S, Bayle M, Lain G, Deleris G (2003) Vascular endothelial cell growth factor (VEGF), an emerging target for cancer chemotherapy. Curr Med Chem Anticancer Agents 3:95–117

  44. 44.

    Reuter S, Gupta SC, Chaturvedi MM, Aggarwal BB (2010) Oxidative stress, inflammation, and cancer: how they linked? Free Radic Biol Med 49:1603–1616

  45. 45.

    Storz P (2005) Reactive oxygen species in tumor progression. Front Biosci 10:1881–1896

  46. 46.

    Rochlitz CF, Peter S, Willroth G, de Kant E, Lobeck H, Huhn D (1992) Mutations in the ras protooncogenes are rare events in renal cell cancer. Eur J Cancer 28:333–336

  47. 47.

    Romanenko AM, Morimura K, Kinoshita A, Wanibuchi H, Takahashi S, Zaparin WK, Vinnichenko VI, Vozianov AF, Fukushima Sh (2006) Upregulation of fibroblast growth factor3 and epidermal growth factor receptors, in association with Raf-1, in urothelial dysplasia and carcinoma in situ after the Chernobyl accident. Cancer Sci 97:1168–1174

  48. 48.

    Suzuki K, Kodama S, Watenabe M (2001) Extremely low-dose ionizing radiation causes activation of mitogen-activated protein kinase pathway and enhances proliferation of normal human diploid cells. Cancer Res 61:5396–5401

  49. 49.

    Chandel NS, Maltere E, Goldwasser E, Mathieu CE, Simon MC, Schumacker PT (1998) Mitochondrial reactive oxygen species trigger hypoxia-induced transcription. Proc Natl Acad Sci USA 95:11715–11720

  50. 50.

    MacLennan GT, Bostwick DG (1995) Microvessel density in renal cell carcinoma: lack of prognostic significance. Urol 46:27–30

  51. 51.

    Kavantzas N, Paraskevakou H, Tseleni-Balafouta S, Aroni K, Athanassiades P, Agrogiannis G, Patsouris E (2007) Association between microvessel density and histologic grade in renal cell carcinomas. Pathol Oncoly Res 13:145–148

  52. 52.

    Puscasiu D, Tatu C, Tatu RF, Potencz E, Popescu R, Muntean I, Verdes D (2011) The significance of angiogenesis and tumoral proliferation in renal cell carcinoma. Rom J Morphol Embryol 52:369–372

Download references


This study is supported by grants: Conselleria Sanitat Comunitat Valenciana, ref. no. 909/2007 and Fundación Instituto Valenciano de Oncología (IVO) Valencia. Spain.

Conflict of interest statement

We declare that we have no conflict of interest.

Author information

Correspondence to A. Llombart-Bosch.

Additional information

Grant for temporary stay for invited professors, number UV-ESTPC-07-393, Valencia University, Spain.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Romanenko, A.M., Ruiz-Saurí, A., Morell-Quadreny, L. et al. Microvessel density is high in clear-cell renal cell carcinomas of Ukrainian patients exposed to chronic persistent low-dose ionizing radiation after the Chernobyl accident. Virchows Arch 460, 611–619 (2012).

Download citation


  • Conventional renal cell carcinoma
  • Ionizing radiation
  • Angiogenesis
  • Microvessel density
  • CD31antibody