Clin Exp Otorhinolaryngol.  2016 Sep;9(3):270-277. 10.21053/ceo.2015.01305.

The Expression and Relationship of CD68-Tumor-Associated Macrophages and Microvascular Density With the Prognosis of Patients With Laryngeal Squamous Cell Carcinoma

• Sun S
• Pan X
• Zhao L
• Zhou J
• Wang H
• Sun Y

• KMID: 2353627
• DOI: http://doi.org/10.21053/ceo.2015.01305

Abstract

OBJECTIVES
We sought to identify the expression of CD68-tumor-associated macrophages (TAMs) and CD34-microvascular density (MVD) in laryngeal squamous cell carcinoma (LSCC), to study the relationship with clinical pathological parameters and to determine whether their expression is predictive of disease.
METHODS
Pathologically confirmed 45 LSCC tissue and 20 peritumoral non-tumor tissue were examined. Immunohistochemical studies were used to detect the expression of CD68-TAMs and CD34-MVD.
RESULTS
The positive expression rate of CD68 in LSCC tissue was 82% (37/45), which was higher than the 10% (2/20) expression rate of the peritumoral tissue (P<0.05). The CD34-MVD positive expression rate in the LSCC tissue was 26.5±6.4, which obviously higher than 12.2±4.0 expression rate of the peritumoral tissue (P<0.05). The positive expression rates of both CD68 and CD34-MVD were higher in the lymph node metastasis (LNM) positive group than in the LNM negative group. The expression of CD68 had positive correlation with CD34-MVD. The 5-year disease-free survival rate in the group with the low CD68 expression was significantly higher than that in the group with high CD68 expression (76% vs. 42%, respectively).
CONCLUSION
The high expression of CD68-TAMs in LSCC and its positive correlation with CD34-MVD illustrates that both play an important role in promoting the metastasis and angiogenesis of this cancer. Their expression was also positively correlated with the prognoses of these patients, suggesting that they could be used as important prognostic markers for LSCC.

Keyword

Laryngeal Neoplasms; Tumor-Associated Macrophages; CD; Microvascular Density

MeSH Terms

Carcinoma, Squamous Cell*
Disease-Free Survival
Epithelial Cells*
Humans
Laryngeal Neoplasms
Lymph Nodes
Macrophages*
Neoplasm Metastasis
Prognosis*

Figure

• 

  Fig. 1. (A) Expression of CD68 in laryngeal squamous cell carcinoma SP (×200). (B) Expression of CD68 in laryngeal squamous cell carcinoma SP (×400). (C) Little or no expression in adjacent non-caner normal tissues SP (×200). (D) Expression of CD34 in tumor nests edge zone SP (×200). (E) Expression of CD34 in Laryngeal squamous cell carcinoma of the inside edge of tumor tissue stroma SP (×200). SP, streptavidin-perosidase.

• 

  Fig. 2. The survival curves in the low infiltration area (low expression zone) and high infiltration area (highly expression area) of CD68 in laryngeal squamous carcinoma. The average survival time was 106.1±10.2 months in CD68 low-expression area, 5-year survival rate was 76% (16/21). The average survival time was 56.3±9.1 months in the high-expression area, 5-year survival rate was 42% (10/24). There was statistical significance between two by log-rank method (P=0.020).

Reference

1. Zhou L, Yang K, Andl T, Wickett RR, Zhang Y. Perspective of targeting cancer-associated fibroblasts in melanoma. J Cancer. 2015; Jun. 6(8):717–26.

2. Weinberg RA. Twisted epithelial-mesenchymal transition blocks senescence. Nat Cell Biol. 2008; Sep. 10(9):1021–3.

3. Cavallo F, De Giovanni C, Nanni P, Forni G, Lollini PL. 2011: the immune hallmarks of cancer. Cancer Immunol Immunother. 2011; Mar. 60(3):319–26.

4. Jang JY, Lee JK, Jeon YK, Kim CW. Exosome derived from epigallocatechin gallate treated breast cancer cells suppresses tumor growth by inhibiting tumor-associated macrophage infiltration and M2 polarization. BMC Cancer. 2013; Sep. 13:421.

5. Duray A, Demoulin S, Hubert P, Delvenne P, Saussez S. Immune suppression in head and neck cancers: a review. Clin Dev Immunol. 2010; 2010:701657.

6. Bostrom MM, Irjala H, Mirtti T, Taimen P, Kauko T, Algars A, et al. Tumor-Associated macrophages provide significant prognostic information in urothelial bladder cancer. PLoS One. 2015; Jul. 10(7):e0133552.

7. Li H, Zhang J, Chen SW, Liu LL, Li L, Gao F, et al. Cancer-associated fibroblasts provide a suitable microenvironment for tumor development and progression in oral tongue squamous cancer. J Transl Med. 2015; Jun. 13:198.

8. Kim S, Cho SW, Min HS, Kim KM, Yeom GJ, Kim EY, et al. The expression of tumor-associated macrophages in papillary thyroid carcinoma. Endocrinol Metab (Seoul). 2013; Sep. 28(3):192–8.

9. Pirila E, Vayrynen O, Sundquist E, Pakkila K, Nyberg P, Nurmenniemi S, et al. Macrophages modulate migration and invasion of human tongue squamous cell carcinoma. PLoS One. 2015; Mar. 10(3):e0120895.

10. Boas DS, Takiya CM, Gurgel CA, Cabral MG, Santos JN. Tumor-infiltrating macrophage and microvessel density in oral squamous cell carcinoma. Braz Dent J. 2013; 24(3):194–9.

11. Lin JY, Li XY, Tadashi N, Dong P. Clinical significance of tumor-associated macrophage infiltration in supraglottic laryngeal carcinoma. Chin J Cancer. 2011; Apr. 30(4):280–6.

12. Klimp AH, Hollema H, Kempinga C, van der Zee AG, de Vries EG, Daemen T. Expression of cyclooxygenase-2 and inducible nitric oxide synthase in human ovarian tumors and tumor-associated macrophages. Cancer Res. 2001; Oct. 61(19):7305–9.

13. Dineen SP, Lynn KD, Holloway SE, Miller AF, Sullivan JP, Shames DS, et al. Vascular endothelial growth factor receptor 2 mediates macrophage infiltration into orthotopic pancreatic tumors in mice. Cancer Res. 2008; Jun. 68(11):4340–6.

14. Ding T, Xu J, Wang F, Shi M, Zhang Y, Li SP, et al. High tumor-infiltrating macrophage density predicts poor prognosis in patients with primary hepatocellular carcinoma after resection. Hum Pathol. 2009; Mar. 40(3):381–9.

15. Green CE, Liu T, Montel V, Hsiao G, Lester RD, Subramaniam S, et al. Chemoattractant signaling between tumor cells and macrophages regulates cancer cell migration, metastasis and neovascularization. PLoS One. 2009; Aug. 4(8):e6713.

16. Noy R, Pollard JW. Tumor-associated macrophages: from mechanisms to therapy. Immunity. 2014; Jul. 41(1):49–61.

17. Kuribayashi K, Motoyama S, Sasaki T, Sato Y, Yoshino K, Wakita A, et al. C-reactive protein reduces the relative number of tumor-associated M2 macrophages and intratumoral angiogenesis in mice. Tohoku J Exp Med. 2014; 233(4):249–55.

18. Downey CM, Aghaei M, Schwendener RA, Jirik FR. DMXAA causes tumor site-specific vascular disruption in murine non-small cell lung cancer, and like the endogenous non-canonical cyclic dinucleotide STING agonist, 2’3’-cGAMP, induces M2 macrophage repolarization. PLoS One. 2014; Jun. 9(6):e99988.

19. Riabov V, Gudima A, Wang N, Mickley A, Orekhov A, Kzhyshkowska J, et al. Role of tumor associated macrophages in tumor angiogenesis and lymphangiogenesis. Front Physiol. 2014; 5:75.

20. Zhang B, Zhang Y, Zhao J, Wang Z, Wu T, Ou W, et al. M2-polarized macrophages contribute to the decreased sensitivity of EGFR-TKIs treatment in patients with advanced lung adenocarcinoma. Med Oncol. 2014; Aug. 31(8):127.

21. Jung KY, Cho SW, Kim YA, Kim D, Oh BC, Park DJ, et al. Cancers with higher density of tumor-associated macrophages were associated with poor survival rates. J Pathol Transl Med. 2015; Jul. 49(4):318–24.

22. Guo C, Buranych A, Sarkar D, Fisher PB, Wang XY. The role of tumor-associated macrophages in tumor vascularization. Vasc Cell. 2013; Dec. 5(1):20.

23. Baeriswyl V, Christofori G. The angiogenic switch in carcinogenesis. Semin Cancer Biol. 2009; Oct. 19(5):329–37.

24. Lin EY, Pollard JW. Tumor-associated macrophages press the angiogenic switch in breast cancer. Cancer Res. 2007; Jun. 67(11):5064–6.

25. Watari K, Shibata T, Kawahara A, Sata K, Nabeshima H, Shinoda A, et al. Tumor-derived interleukin-1 promotes lymphangiogenesis and lymph node metastasis through M2-type macrophages. PLoS One. 2014; Jun. 9(6):e99568.

26. Bolat F, Kayaselcuk F, Nursal TZ, Yagmurdur MC, Bal N, Demirhan B. Microvessel density, VEGF expression, and tumor-associated macrophages in breast tumors: correlations with prognostic parameters. J Exp Clin Cancer Res. 2006; Sep. 25(3):365–72.

27. Zhang M, He Y, Sun X, Li Q, Wang W, Zhao A, et al. A high M1/M2 ratio of tumor-associated macrophages is associated with extended survival in ovarian cancer patients. J Ovarian Res. 2014; Feb. 7:19.

28. Lin CN, Wang CJ, Chao YJ, Lai MD, Shan YS. The significance of the co-existence of osteopontin and tumor-associated macrophages in gastric cancer progression. BMC Cancer. 2015; Mar. 15:128.

29. Chen SJ, Zhang QB, Zeng LJ, Lian GD, Li JJ, Qian CC, et al. Distribution and clinical significance of tumour-associated macrophages in pancreatic ductal adenocarcinoma: a retrospective analysis in China. Curr Oncol. 2015; Feb. 22(1):e11–9.

30. Colvin EK. Tumor-associated macrophages contribute to tumor progression in ovarian cancer. Front Oncol. 2014; Jun. 4:137.

31. Deng R, Wang SM, Yin T, Ye TH, Shen GB, Li L, et al. Dimethyl sulfoxide suppresses mouse 4T1 breast cancer growth by modulating tumor-associated macrophage differentiation. J Breast Cancer. 2014; Mar. 17(1):25–32.

32. Ohri CM, Shikotra A, Green RH, Waller DA, Bradding P. Macrophages within NSCLC tumour islets are predominantly of a cytotoxic M1 phenotype associated with extended survival. Eur Respir J. 2009; Jan. 33(1):118–26.