The evaluation of lymphatic vessel density and microvessel density in laryngeal squamous cell carcinoma

Abstract

Aim: Metastasis to the cervical lymph nodes in laryngeal carcinomas is a factor reducing survival significantly.  The present study aimed to reveal the association of lymph node metastasis with lymphatic vessel density and microvessel density in the intratumoral and extra-tumoral areas of laryngeal squamous cell carcinoma.

Methods: Eighty-six cases diagnosed with laryngeal squamous cell carcinoma in Pathology Department of Erciyes University Faculty of Medicine between 2000-2006 were included in the present study. Lymphatic vessel density and microvessel density were assessed with D2-40 which is an immunohistochemical marker in intratumoral and extra-tumoral areas and CD34, respectively. The results were compared with tumor grade, tumor localization, and lymph node metastasis.

Results: Intratumoral and extra tumoral lymphatic vessel density value were 8.93±12.5 and 24.1±20.1, respectively (p=0.001). Mean intra tumoral microvessel density was calculated as 217.53±89.8 while extra tumoral microvessel density was calculated as 330.43±92.4 (p=0.001). Intratumoral lymphatic vessel density value was higher in the well-differentiated tumors compared to the poorly differentiated tumors but no significant result was obtained (p=0.100). Extra tumoral microvessel density value was found to be significantly higher in the poorly differentiated tumors (p=0.05). Intratumoral microvessel density value was significantly lower in tumors with lymph node metastasis (p=0.028). Extra-tumoral lymphatic vessel density value was higher in both groups with and without lymph node metastasis, however, no statistically significant result could be obtained (p=0.084).

Conclusion: In the present study, intratumoral microvessel density was higher in well differentiated tumors whereas extra tumoral microvessel density was determined to be higher in poorly differentiated cases. No significant association was noted between lymphatic vessel density and differentiation. Extra tumoral and intra tumoral microvessel density values were found to be higher in the group without metastasis.  No association was detected between metastasis and lymphatic vessel density value.

We suggest that assessing lymphatic vessel density with the co-administration of D2-40 and CD34 may be more important for early detection of metastasis.

Keywords

• : D2-40,angiogenesis,CD34,larynx,squamous cell carcinoma

Laringeal skuamöz hücreli karsinomlarda lenfatik damar yoğunluğu ve mikrodamar yoğunluğunun değerlendirilmesi

Öz

Amaç: Larinks karsinomlarında boyun lenf nodlarına metastaz, sağ kalımı önemli oranda azaltan bir etkendir.  Bu çalışmanın amacı; larinksin skuamöz hücreli tümörlerinde tümör içi ve tümör dışı alanlarda lenfatik damar yoğunluğu ve mikrodamar yoğunluğunun lenf nodu metastazı ile ilişkisini ortaya koymaktır.

Yöntemler: Çalışmamızda 2000-2006 yılları arasında Erciyes  Üniversitesi Tıp Fakültesi Patoloji Ana Bilim Dalı’nda larinks skuamöz hücreli karsinom tanısı almış 86 olgu incelemeye alındı. Tümör içi ve tümör dışı alanlarda immünhistokimyasal bir belirteç olan D2-40 ile lenfatik damar yoğunluğu ve CD34 ile mikrodamar damar yoğunluğu değerlendirildi. Sonuçlar tümör derecesi, tümör yerleşim yeri, lenf nodu metastazı ile karşılaştırıldı.

Bulgular: Olguların tümör içi ve tümör dışı lenfatik damar yoğunluğu sırası ile 8,93±12,5  ve 24,1±20,1 idi (p=0,001). Tümör içi mikrodamar yoğunluğu değeri 217,53±89,8 ve tümör dışı mikrodamar yoğunluğu değeri 330,43±92,4 olarak sayıldı (p=0,001). İyi diferansiye tümörlerde tümör içi lenfatik damar yoğunluğu değeri, iyi diferansiye olmayan tümörlere göre yüksekti, fakat anlamlı sonuç elde edilemedi (p=0,100). Kötü diferansiye tümörlerde tümör dışı mikrodamar yoğunluğu değeri anlamlı olarak yüksek bulundu (p=0,050). Tümör içi mikrodamar yoğunluğu değeri lenf nodu metastazı olan tümörlerde anlamlı olarak düşüktü (p=0,028). Lenf nodu metastazı olan ve olmayan her iki grupta da tümör dışı lenfatik damar yoğunluğu değeri yüksekti, istatiksel olarak anlamlı bir sonuç elde edilemedi (p=0,084). Sonuç: Bu çalışmada iyi diferansiye tümörlerde tümör içi mikrodamar yoğunluğu ve kötü diferansiye olgularda tümör dışı mikrodamar yoğunluğu daha yüksek idi. Lenfatik damar yoğunluğu değeri ile diferansiasyon arasında bir ilişki saptanmadı. Metastaz yapmayan grupta tümör içi ve tümör dışı mikrodamar yoğunluğu değeri yüksek bulundu.  Metastaz ile lenfatik damar yoğunluğu değeri arasında bir ilişki izlenmedi.

D2-40 ve CD34 ‘ün birlikte kullanımı ile lenfatik damar yoğunluğunun değerlendirilmesinin, metastazın erken belirlenmesinde daha önemli olabileceğini düşünmekteyiz.

Anahtar Kelimeler

• D2-40,anjiogenez,CD34,larinks,skuamöz hücreli karsinom

References

1. 1. Rosai J. Ackerman's Surgical Pathology (9 ed). China, Elsevier, 2004; pp. 341-7.
2. 2. Barnes L, Evson JW, Reichart P, Sidransky D. Pathology and Genetics of Head and Neck Tumours Lyon, IARC, 2005;108-39.
3. 3. [Pietruszewska W, Kobos J, Grycznski M. Microvessel density and endothelial area in assessment of angiogenesis in patients with laryngeal cancer]. Otolaryngol Pol. 2003;57:5-15.
4. 4. Heymach JV. Angiogenesis and antiangiogenic approaches to sarcomas. Curr Opin Oncol. 2001;13:261-9.
5. 5. Li J, Tang L, Ren Z, Pan Z. Distribution and prognostic significance of microvessel density in supraglottic laryngeal squamous cell carcinomas. Lin Chuang Er Bi Yan Hou Ke Za Zhi. 2004;18:513-5.
6. 6. Sion-Vardy N, Fliss DM, Prinsloo I, Shoham-Vardi I, Benharroch D. Neoangiogenesis in squamous cell carcinoma of the larynx-biological and prognostic associations. Pathol Res Pract. 2001;197:1-5.
7. 7. Pietruszewska W, Niewiadomska H, Kobos J, Jozefowic-Korczynska M, Gryczynski M. Significance of angiogenesis in laryngeal cancer. Otolaryngol Pol. 2000;31:167-70.
8. 8. Yan Gao, Wei-Xia Zhong, Dian-Bin Mu, Yuan YP, Zhang YH, Yu JM, et al. Distributions of Angiogenesis and Lymphangiogenesis in Gastrointestinal Intramucosal Tumors. Ann Surg Oncol. 2008;15:1117-23.

9. 9. William WL Choi, Melinda M Lewis, Diane Lawson, Yin-Goen Q, Birdsong GG, Cotsonis GA, et al. Angiogenic and lymphangiogenic mivrovessel density in breast carcinoma: correlation with clinicopathologic parameters and VEGF-family gene expression. Mod Pathol. 2005;18:143-52.
10. 10. Weidner N, Semple JP, Welch WR, Folkman J. Tumor angiogenesis and metastasis –correlation in invasive breast cancer. N Engl J Med. 1991;324:1-8.
11. 11. Koskinen WJ, Bono P, Leivo I, Vaheri A, Aaltonen LM, Joensuu H. Lymphatic vessel density in vocal cord carcinomas assessed with LYVE-1 receptor expression. Radiother Oncol. 2005;77:172-5.
12. 12. Fernandez MI, Bolenz C, Trojan L, Steidler A, Weiss C, Alken P et al. Prognostic implications of lymphangiogenesis in muscle-ınvasive transitional celi carcinoma of the bladder. Eur Urol. 2008;53:571-8.
13. 13. Gomaa AHA, Yaar M, Bhawan J. Cutaneous ımmunoreactivity of D2-40 antibody beyond the lymphatics. Am J Dermathopathol. 2007;29:18-21.
14. 14. Chu AY, Litzky LA, Pasha TL, Acs G, Zhang PJ. Utility of D2-40, a novel mesothelial marker, in the diagnosis of malignant mesothelioma. Mod Pathol. 2005;18:105-10.
15. 15. Ge Yan, Xiao-Yan Zhou, San-Jun Cai, Zhang GH, Peng JJ, Du X. Lymphangiogenic and angiogenic microvessel density in human primary sporadic colorectal carcinoma. World J Gastroenterol. 2008;14:101-7.
16. 16. Longatto Filho A, Oliveira TG, Pinheiro C, de Carvalho MB, Curioni OA, Mercante AM, et al. How useful is the assessment of lymphatic vascular density in oral carcinoma prognosis? World J Surg Oncol. 2007;5:140.
17. 17. Arnaout-Alkarain A, Kahn HJ, Narod SA, Sun PA, Marks AN. Significance of lymph vessel invasion identified by the endothelial lymphatic marker D2-40 in node negative breast cancer. Mod Pathol. 2007;20:183-91.
18. 18. Mimura T, Ito A, Sakuma T, Ohbayashi C, Yoshimura M, Tsubota N, et al. Novel Marker D2-40, Combined With Calretinin, CEA, and TTF-1. Cancer. 2007;109:933-98.
19. 19. Joensvu H. Gastrointestinal stromal tumor (GIST). Ann Oncol. 2006;17:280-6.
20. 20. Ito M, Moriya T, Ishida T, Usami S, Kasajima A, Sasano H, et al. Sinificance of pathological evaluation for lymphatic vessel invasion in invasive breast cancer. Breast Cancer. 2007;14:381-7.
21. 21. Saad RS, Kordunsky L, Liu YL, Denning KL, Kandil HA, Silverman JF. Lymphatic microvessel density as prognostic marker in colorectal cancer. Mod Pathol. 2006;19:1317-23.
22. 22. El-Gohary YM, Metwally G, Saad RS, Robinson MJ, Mesko T, Poppiti RJ. Prognostic significance of intratumoral and peritumoral lymphatic density and blood vessel density in invasive breast carcinoma. Am J Clin Pathol. 2008;129:578-86.
23. 23. Hobbs CG, Birchall MA. Human papillomavirus infection in the etiology of laryngeal carcinoma. Curr Opin Otolaryngol Head Neck Surg. 2004;12:88-92.
24. 24. Goldenberg D, Benoit NE, Begum S, Westra WH, Cohen Y, Koch WM, et al. Epstein-Barr virus in head and neck cancer assessed by quantitative polymerase chain reaction. Laryngoscope. 2004;114:1027-31.
25. 25. Rabbett WF. Juvenile laryngeal papillomatosis. The relation of irradiation to malignant degeneration in this disease. Ann Otol Rhinol Laryngol. 1965;74:1149-63.
26. 26. Berkower AS, Biller HF. Head and neck cancer associated with Bloom's syndrome. Laryngoscope. 1988;98:746-8.
27. 27. Potzsch C, Voigtlander T, Lubbert M. p53 Germline mutation in a patient with Li- Fraumeni Syndrome and three metachronous malignancies. J Cancer Res Clin Oncol. 2002;128:456-60.
28. 28. Xuan M, Fang YR, Wato M, Hata S, Tanaka A. Immunohistochemical colocalization of lymphatics and blood vessels in oral squamous cell carcinomas. J Oral Pathol Med. 2005;34: 334-9.
29. 29. Kyzas PA, Stefanou D, Agnantis NJ. COX-2 expression correlates with VEGF-Cand lymph node metastases in patients with head and neck squamous cell carcinoma. Mod Pathol. 2005;18:153-60.
30. 30. Saad RS, Kordunsky L, Liu YL, Denning KL, Kandil HA, Silverman JF. Lymphatic microvessel density as prognostic marker in colorectal cancer. Mod Pathol. 2006;19:1317-23.
31. 31. Naik VR, Jaafar H, Seng CE. Lymphatic channel density in colorectal adenocarcinoma. Indian J Pathol Microbiol. 2010;53:12-4.
32. 32. Gombos Z, Xu X, Chu CS, Zhang P, Acs G. Peritumoral lymphatic vessel density and vascular endothelial grovvth factor C expression in early-stage squamous cell carcinoma of the uterine cervix. Clin Cancer Res. 2005;11:8364-71.
33. 33. Coşkun U, Akyürek N, Dursun A, Yamaç D. Peritumoral lymphatic microvessel density associated with tumor progression and poor prognosis in gastric carcinoma. J Surg Res. 2009;3:1-6.
34. 34. Roma AA, Magi-Galluzzi C, Kral MA, Jin TT, Klein E A, Zhou M. Peritumoral lymphatic invasion is associated with regional lymph node metastases in prostate adenocarcinoma. Mod Pathol. 2006;19:392-8.
35. 35. Huangfu H, Kong W, Gong S, Wang B, Zhang C. Study on relationship between intratumoral lymphangiogenesis of laryngeal squamous cell carcinoma and lymphatic metastasis. Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2008;22:385-8.
36. 36. Mannelqvist M, Stefansson I, Salvesen HB, Akslen LA. Importance of tumour cell invasion in blood and lymphatic vasculature among patients with endometrial carcinoma. Histopathology. 2009;54:174-83.
37. 37. Sternberg SS. Histology for Pathologists. In:Mills SE, Fecher ER. Larynx and Pharynx. 2nd ed. Philadelphia:Lippincott-Raven, 1997; pp. 391-403.
38. 38. Tomik J, Skladzien J, Modrzejeweski M. Evaluation of cervical lymph node metastasis of 1400 patients with cancer of the larynx. Auris Nasus Larynx. 2001;28:233-40.
39. 39. Rodrigo JP, Cabanillas R, Chiara MD, Garcia Pedrero J, Astudillo a, Suarez Nieto C. Prognostic significance of angiogenesis in surgically treated supraglottic squamous cell carcinomas of the larynx. Acta Otorrinolaringol Esp. 2009;60: 272-7.
40. 40. Kyzas PA, Stefanou D, Agnantis NJ. COX-2 expression correlates with VEGF-C and lymph node metastases in patients with head and neck squamous celi carcinoma. Mod Pathol. 2005;18:153-60.
41. 41. Franchi A, Gallo O, Massi D, Baroni G, Santucci M. Tumor Lymphangiogenesis in Head and Neck Squamous Celi Carcinoma. Cancer. 2004;101:973-8.
42. 42. Seppälä M, Pohjola K, Laranne J, Rautiainen M, Huhtala H, Renkonen R, et al. High relative density of lymphatic vessels predict poor survival in tongue squamous cell carcinoma. Eur Arch Otorhinolaryngol. 2016;273:4515-24.
43. 43. Huse JT, Pasha TL, Zhang PJ. D2-40 functions as an effective chonroid marker distinguishing true chondroid tumors from chordoma. Acta Neuropathol 2007;113:87-94.
44. 44. Chu AY, Litzky LA, Pasha TL, Acs G, Zhang PJ. Utility of D2-40, a novel mesothelial marker, in the diagnosis of malignant mesothelioma. Mod Pathol 2005;18:105-10.
45. 45. Ordonez NG. D2-40 and podaplanin are highly specific and sensitive immunohistochemical markers of epithelioid malignant mesothelioma. Hum Pathol. 2005;36:372-80.
46. 46. Hansen T, Katenkamp K, Bittinger F, Kirkpatrick CJ, Katenkamp D. D2-40 labelingin lymphangiomyoma/lymphangiomyomatosis of soft tissue: further evidence of lymphangiogenic tumor histogenesis. Virchows Arch. 2007;450:449-53.
47. 47. Iwakiri S, Nagai S, Katakura H, Takenaka K, Date H, Wada H, et al. D2-40 Positive Lymphatic Vessel Density Is a Poor Prognostic Factor in Squamous Cell Carcinoma Of the Lung. Ann Surg Oncol 2009;16:1678-85.