Abstract
The growth, invasiveness, and metastasis of human cancers are not only determined by the cancer cells but also by their microenvironment. The purpose of this study was to extend our previous studies and to examine the cellular changes in tumor microenvironment (stroma) of esophageal squamous cell carcinomas (ESCCs). The proliferative activity, cellular components, and angiogenesis status in different compartments (non-tumor stroma, tumor stroma, and tumor periphery stroma) of ESCCs were evaluated by immunohistochemistry. The results revealed a hyperproliferative rate labeled by Ki-67 in stromal cells in tumor area as compared with that in stromal cells in non-tumor area, which resulted in the increased densities of myofibroblasts (labeled by smooth muscle actin (SMA)-alpha), lymphocytes (labeled by CD3), macrophages (labeled by CD68), and the activation of angiogenesis characterized by increased microvessel density (MVD) and the increased expression of the proangiogenic factors (vascular endothelial growth factor and interleukin 8) in the tumor stroma. Further analysis showed that the changes of stromal cell density were more significant in the area of periphery tumor stroma than that of stroma between tumor nests. Most cellular changes were significantly associated with lymph node involvement. Double immunohistochemistries with PCNA/CD3, PCNA/CD68, and PCNA/SMA-alpha revealed that these cells present in the ESCC tumor stroma had a proliferative capacity. The cells present in the tumor microenvironment of ESCCs were greatly activated, suggesting that microenvironmental components may be involved in the cancer growth and progression.
Similar content being viewed by others
Abbreviations
- PCNA:
-
Proliferating cell nuclear antigen
- MVD:
-
Microvessel density
- IL:
-
Interleukin
- COX-2:
-
Cyclooxygenase-2
- IDO:
-
Indoleamine 2, 3-dioxygenase
- IHC:
-
Immunohistochemistry
- ESCC:
-
Esophageal squamous cell carcinoma
References
Ke L. Mortality and incidence trends from esophagus cancer in selected geographic areas of China circa 1970–90. Int J Cancer. 2002;102:271–4.
Song PI, Liang H, Fan JH, Wei WQ, Wang GQ, Qiao YL. Long-term survival after esophagectomy for early esophageal squamous cell carcinoma in Linxian, China. J Surg Oncol. 2011;104:176–80.
Whiteside TL. The role of immune cells in the tumor microenvironment. Cancer Treat Res. 2006;130:103–24.
Schottelius AJ, Dinter H. Cytokines, NF-kappaB, microenvironment, intestinal inflammation and cancer. Cancer Treat Res. 2006;130:67–87.
Gholamin M, Moaven O, Memar B, Farshchian M, Naseh H, Malekzadeh R, Sotoudeh M, Rajabi-Mashhadi MT, Forghani MN, Farrokhi F, Abbaszadegan MR. Overexpression and interactions of interleukin-10, transforming growth factor beta, and vascular endothelial growth factor in esophageal squamous cell carcinoma. World J Surg. 2009;33:1439–45.
Hsia JY, Chen JT, Chen CY, Hsu CP, Miaw J, Huang YS, Yang CY. Prognostic significance of intratumoral natural killer cells in primary resected esophageal squamous cell carcinoma. Chang Gung Med J. 2005;28:335–40.
Mukherjee S, Roth MJ, Dawsey SM, Yan W, Rodriguez-Canales J, Erickson HS, Hu N, Goldstein AM, Taylor PR, Richardson AM, Tangrea MA, Chuaqui RF, Emmert-Buck MR. Increased matrix metalloproteinase activation in esophageal squamous cell carcinoma. J Transl Med. 2010;8:91.
Noma K, Smalley KS, Lioni M, Naomoto Y, Tanaka N, El-Deiry W, King AJ, Nakagawa H, Herlyn M. The essential role of fibroblasts in esophageal squamous cell carcinoma-induced angiogenesis. Gastroenterology. 2008;134:1981–93.
Yuan A, Liu J, Liu Y, Bjornsen T, Varro A, Cui G. Immunohistochemical examination of gastrin, gastrin precursors, and gastrin/CCK-2 receptor in human esophageal squamous cell carcinomas. Pathol Oncol Res. 2008;14:449–55.
Zhang C, Fu L, Fu J, Hu L, Yang H, Rong TH, Li Y, Liu H, Fu SB, Zeng YX, Guan XY. Fibroblast growth factor receptor 2-positive fibroblasts provide a suitable microenvironment for tumor development and progression in esophageal carcinoma. Clin Cancer Res. 2009;15:4017.
Kubota Y, Kaneko K, Konishi K, Ito H, Yamamoto T, Katagiri A, Muramoto T, Yano Y, Kobayashi Y, Oyama T, Kushima M, Imawari M. The onset of angiogenesis in a multistep process of esophageal squamous cell carcinoma. Front Biosci. 2009;14:3872–8.
Noguchi T, Takeno S, Shibata T, Uchida Y, Yokoyama S, Muller W. VEGF-C expression correlates with histological differentiation and metastasis in squamous cell carcinoma of the esophagus. Oncol Rep. 2002;9:995–9.
Yu L, Wu WK, Li ZJ, Li HT, Wu YC, Cho CH. Prostaglandin E(2) promotes cell proliferation via protein kinase C/extracellular signal regulated kinase pathway-dependent induction of c-Myc expression in human esophageal squamous cell carcinoma cells. Int J Cancer. 2009;125:2540–6.
Bergmann C, Strauss L, Zeidler R, Lang S, Whiteside TL. Expansion of human T regulatory type 1 cells in the microenvironment of cyclooxygenase 2 overexpressing head and neck squamous cell carcinoma. Cancer Res. 2007;67:8865–73.
Moussai D, Mitsui H, Pettersen JS, Pierson KC, Shah KR, Suarez-Farinas M, Cardinale IR, Bluth MJ, Krueger JG, Carucci JA. The human cutaneous squamous cell carcinoma microenvironment is characterized by increased lymphatic density and enhanced expression of macrophage-derived VEGF-C. J Investig Dermatol. 2011;131:229–36.
Thode C, Jorgensen TG, Dabelsteen E, Mackenzie I, Dabelsteen S. Significance of myofibroblasts in oral squamous cell carcinoma. J Oral Pathol Med: Official Publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology. 2011;40:201–7.
Rao SK, Pavicevic Z, Du Z, Kim JG, Fan M, Jiao Y, Rosebush M, Samant S, Gu W, Pfeffer LM, Nosrat CA. Pro-inflammatory genes as biomarkers and therapeutic targets in oral squamous cell carcinoma. J Biol Chem. 2010;285:32512–21.
Lewis CE, Pollard JW. Distinct role of macrophages in different tumor microenvironments. Cancer Res. 2006;66:605–12.
Zips D, Eicheler W, Bruchner K, Jackisch T, Geyer P, Petersen C, van der Kogel AJ, Baumann M. Impact of the tumour bed effect on microenvironment, radiobiological hypoxia and the outcome of fractionated radiotherapy of human FaDu squamous-cell carcinoma growing in the nude mouse. Int J Radiat Biol. 2001;77:1185–93.
Guo SJ, Lin DM, Li J, Liu RZ, Zhou CX, Wang DM, Ma WB, Zhang YH, Zhang SR. Tumor-associated macrophages and CD3-zeta expression of tumor-infiltrating lymphocytes in human esophageal squamous-cell carcinoma. Dis Esophagus. 2007;20:107–16.
Watanabe M, Kono K, Kawaguchi Y, Mizukami Y, Mimura K, Maruyama T, Izawa S, Fujii H. NK cell dysfunction with down-regulated CD16 and up-regulated CD56 molecules in patients with esophageal squamous cell carcinoma. Dis Esophagus. 2010;23:675–81.
Liu J, Lu G, Li Z, Tang F, Liu Y, Cui G. Distinct compartmental distribution of mature and immature dendritic cells in esophageal squamous cell carcinoma. Pathol Res Pract. 2010;206:602–6.
Liu J, Lu G, Tang F, Liu Y, Cui G. Localization of indoleamine 2,3-dioxygenase in human esophageal squamous cell carcinomas. Virchows Arch. 2009;455:441–8.
Ishigami S, Natsugoe S, Matsumoto M, Okumura H, Sakita H, Nakashima S, Takao S, Aikou T. Clinical implications of intratumoral dendritic cell infiltration in esophageal squamous cell carcinoma. Oncol Rep. 2003;10:1237–40.
Hu WM, Li L, Jing BQ, Zhao YS, Wang CL, Feng L, Xie YE. Effect of S1P5 on proliferation and migration of human esophageal cancer cells. World J Gastroenterol: WJG. 2010;16:1859–66.
Cui G, Koh TJ, Chen D, Zhao CM, Takaishi S, Dockray GJ, Varro A, Rogers AB, Fox JG, Wang TC. Overexpression of glycine-extended gastrin inhibits parietal cell loss and atrophy in the mouse stomach. Cancer Res. 2004;64:8160–6.
Cui G, Yuan A, Vonen B, Florholmen J. Progressive cellular response in the lamina propria of the colorectal adenoma–carcinoma sequence. Histopathology. 2009;54:550–60.
Cui G, Goll R, Olsen T, Steigen SE, Husebekk A, Vonen B, Florholmen J. Reduced expression of microenvironmental Th1 cytokines accompanies adenomas–carcinomas sequence of colorectum. Cancer Immunol Immunother. 2007;56:985–95.
Adegboyega PA, Ololade O, Saada J, Mifflin R, Di Mari JF, Powell DW. Subepithelial myofibroblasts express cyclooxygenase-2 in colorectal tubular adenomas. Clin Cancer Res. 2004;10:5870–9.
Weidner N, Semple JP, Welch WR, Folkman J. Tumor angiogenesis and metastasis—correlation in invasive breast carcinoma. N Engl J Med. 1991;324:1–8.
De Wever O, Mareel M. Role of tissue stroma in cancer cell invasion. J Pathol. 2003;200:429–47.
Yamada M, Suzu S, Tanaka-Douzono M, Wakimoto N, Hatake K, Hayasawa H, Motoyoshi K. Effect of cytokines on the proliferation/differentiation of stroma-initiating cells. J Cell Physiol. 2000;184:351–5.
Yu P, Fu YX. Tumor-infiltrating T lymphocytes: friends or foes? Lab Investig; A Journal of Technical Methods and Pathology. 2006;86:231–45.
O’Sullivan C, Lewis CE. Tumour-associated leucocytes: friends or foes in breast carcinoma. J Pathol. 1994;172:229–35.
Sadanaga N, Kuwano H, Watanabe M, Maekawa S, Mori M, Sugimachi K. Local immune response to tumor invasion in esophageal squamous cell carcinoma. The expression of human leukocyte antigen-DR and lymphocyte infiltration. Cancer. 1994;74:586–91.
Kalluri R, Zeisberg M. Fibroblasts in cancer. Nat Rev Cancer. 2006;6:392–401.
Rasanen K, Vaheri A. Activation of fibroblasts in cancer stroma. Exp Cell Res. 2010;316:2713–22.
Tsuzuki S, Ota H, Hayama M, Sugiyama A, Akamatsu T, Kawasaki S. Proliferation of alpha-smooth muscle actin-containing stromal cells (myofibroblasts) in the lamina propria subjacent to intraepithelial carcinoma of the esophagus. Scand J Gastroenterol. 2001;36:86–91.
Hofmeister V, Schrama D, Becker JC. Anti-cancer therapies targeting the tumor stroma. Cancer Immunol Immunother. 2008;57:1–17.
Akutsu Y, Hanari N, Yusup G, Komatsu-Akimoto A, Ikeda N, Mori M, Yoneyama Y, Endo S, Miyazawa Y, Matsubara H. COX2 expression predicts resistance to chemoradiotherapy in esophageal squamous cell carcinoma. Ann Surg Oncol. 2011;18:2946–51.
Krzystek-Korpacka M, Matusiewicz M, Diakowska D, Grabowski K, Blachut K, Konieczny D, Kustrzeba-Wojcicka I, Terlecki G, Banas T. Elevation of circulating interleukin-8 is related to lymph node and distant metastases in esophageal squamous cell carcinomas—implication for clinical evaluation of cancer patient. Cytokine. 2008;41:232–9.
Li Z, Liu J, Tang F, Liu Y, Waldum HL, Cui G. Expression of non-mast cell histidine decarboxylase in tumor-associated microvessels in human esophageal squamous cell carcinomas. APMIS. 2008;116:1034–42.
Acknowledgments
We express our sincere gratitude to Ms. Dana Frederick, Department of Cell Biology, University of Massachusetts Medical School for manuscript proofreading.
Conflicts of interest
None
Author information
Authors and Affiliations
Corresponding author
Additional information
Jinzhong Liu and Zhenfeng Li contributed equally to this study.
Rights and permissions
About this article
Cite this article
Liu, J., Li, Z., Cui, J. et al. Cellular changes in the tumor microenvironment of human esophageal squamous cell carcinomas. Tumor Biol. 33, 495–505 (2012). https://doi.org/10.1007/s13277-011-0281-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13277-011-0281-3