ONLINE MANUSCRIPT SUBMISSION
Editorial Board | Instructions for Authors | Main Page | Table of Contents | Archive | Search
The Turkish Journal of Gastroenterology
Turk J Gastroenterol 2011; 22 (4): 363-368
[ Summary ] [ PDF ] [ Similar Articles ] [ Mail to Author ] [ Mail to Editor ]
Fascin expression and its potential significance in gastrointestinal stromal tumors
Ayhan ÖZCAN1, Yıldırım KARSLIOĞLU1, Armağan GÜNAL1, A. Hakan ÇERMİK2, Bülent KURT1, Önder ÖNGÜRÜ1
Department of 1Pathology, Gülhane Military Medical Academy, School of Medicine, Ankara
Department of 2Pathology, Military Hospital, Etimesgut, Ankara
Keywords: Gastrointestinal stromal tumor, immunohistochemistry, fascin, tumor progression, prognosis.
Summary
Background/aims: Fascin is an actin-bundling protein that is important in cell motility. Fascin expression has been shown to have a potential role in tumor progression for some epithelial tumors. However, there are only a few studies related to its expression in mesenchymal tumors. We investigated fascin expression in gastrointestinal stromal tumors. Methods: Thirty gastrointestinal stromal tumors, which were very low (n=6), low (n=2), moderate (n=4), and high (n=18) risk, constituted our series. Immunohistochemical expression of fascin was studied in all cases. Results: Immunoreactivity was observed in only five cases, all of which were in the high-risk group. The remaining cases (25/30) showed no immunoreactivity, and the difference did not seem statistically important (p=0.261). Fascin expression was stronger in epithelioid cells than spindle-shaped cells (p=0.003). In addition, gastrointestinal stromal tumors in the small bowel showed higher fascin expression than those in the other localizations (p=0.043). Conclusions: It seems that anatomic localization and the tumor cell type of gastrointestinal stromal tumors show statistically significant differences with regard to fascin expression. Although our series is limited, we think that fascin should be considered as a marker that is worthy of further study for its potential usage as a prognostic indicator in gastrointestinal stromal tumors.
  • Top
  • Summary
  • Introduction
  • Materials And Methods
  • Results
  • Discussion
  • References
  • Introduction
    Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract (1-13). GISTs most commonly arise in the stomach (51%), and affect to a much lesser extent the remaining parts of the gastrointestinal tract as follows: small intestine (36%), colon (7%), rectum (1%), or esophagus (1%).

    Most GISTs are immunoreactive for CD 117 (c-kit) and CD 34, and are typically negative for S-100 protein and desmin (1-14). CD 117 positivity is present in all histological variants including benign and malignant GISTs of different sites. There are only a few reports stating that a small number of GISTs might be negative for CD 117 (12,13). It is known that Ki-67 (MIB-1) expression in a GIST of more than 10% likely indicates poor prognosis (2,9).

    Tumor progression is characterized by both the loss of cell-cell and cell-matrix interactions as well as the invasion of the surrounding tissues by tumor cells. All cells are capable of organizing their protoplasmic actin filaments into cross-linked structures that influence cell shape, proliferation, adhesion, motility, and intracellular signaling pathway. Numerous actin-bundling proteins have been described, such as villin, fimbrin, ezrin, spectrin, filamin, and fascin (15-17).

    Fascin is a 55 kDa cross-linking protein that localizes the core actin bundles of spikes and filopodia at the leading edge of migratory cells (15-30). Fascin promotes the formation of tightly packed parallel bundles with F-actin (16,19,21). There are three forms of fascin: fascin-1 (also known as fascin), fascin-2 (retinal fascin) and fascin-3 (testicular fascin) (16,19,21). Fascin has a role in the organization of actin-based structures that function in cell-cell and cell-matrix interactions and motility. Fascin binds beta-catenin, a molecule that is not only an important part of the cell-cell interaction, but also a major signaling molecule in the Wnt pathway (15). It has been proposed that fascin might be expected to behave like an anti-oncogene because of its reducing effect on the cytoplasmic pool of soluble beta-catenin (15). Fascin is expressed in normal mesenchymal, endothelial, dendritic, and neuronal cells, but not in normal simple columnar epithelia of the biliary duct, breast, colon, ovary, pancreas, and stomach epithelia (16,19). The expression of fascin in epithelial neoplasms, on the other hand, has recently been described. In normal epithelial cells, fascin expression is usually present in a very low amount but is often upregulated in several types of human neoplasms, such as ovarian, breast, pancreatic, lung, skin, colon, stomach, and esophagus tumors (20-30). It is not yet known how and why fascin is upregulated in carcinoma cells. Additionally, it is not clear whether the increased fascin protein expression by carcinomas is related to transcriptional or posttranscriptional events, or if the same mechanism is involved in carcinomas of different organs (19).

    There are a limited number of studies regarding the role of fascin expression in spindle cell tumors (31). While fascin was evaluated as a potential marker of dendritic cell neoplasms, it does not seem to be a useful marker in the diagnosis of other spindle cell tumors (31). We aimed to investigate fascin expression with regard to anatomic localizations, tumor invasiveness, prognostic risk groups, and predominant morphological features or cell types of GISTs.
  • Top
  • Summary
  • Introduction
  • Materials And Methods
  • Results
  • Discussion
  • References
  • Materials And Methods
    A total of 30 GIST cases diagnosed from the resection materials between 1995 and 2008 were retrieved from the archives of the Department of Pathology, Gülhane Military Medical Academy. Our cases were categorized into groups as follows: 6 cases, very low risk; 2 cases, low risk; 4 cases, moderate risk; and 18 cases, high risk (Table 1). The rates of metastases or tumor-related death in GISTs were grouped by Miettinen and Lasota (7) classification according to tumor location, tumor size and mitotic rate. The distribution of our cases according to this new classification is demonstrated in Table 2.

    In order to check and confirm the diagnosis of GIST, an immunohistochemical panel consisting of six antibodies against CD 117, CD 34, smooth muscle actin (SMA), desmin, S-100 protein, and Ki-67 was performed on all cases, and reassessed by one of the participants (A.Ö.) (10). The most suitable paraffin block for fascin immunostaining (FCN01; 1: 100 dilution; NeoMarkers, Fremont, CA, USA) was selected by reexamining 3-15 hematoxylin-eosin (H&E)-stained slides. For demonstrating the fascin expression, standard 5-μm sections were obtained and deparaffinized with xylene and rehydrated in graded alcohols. Positive cases for fascin were also stained by SMA (Ab1; 1: 200 dilution; NeoMarkers, Fremont, CA, USA) and muscle specific antigen (MSA) (HHF35; 1: 100 dilution; NeoMarkers, Fremont, CA, USA). All immunohistochemical staining was carried out using Lab Vision Autostainer 480 (Lab Vision Products, Thermo Fisher Scientific, Fremont, CA, USA).

    Each section was thoroughly examined to assess and record both the intensity and the extension of fascin immunolabeling. A positive result was defined as strong staining in at least 10% of tumor cells. Besides stained neoplastic cells, we noticed individually highlighted cells throughout the tumors. Because of their slightly different morphology and the pattern of staining, we interpreted these cells as the entrapped dendritic cells. Dendritic cells have cytoplasmic processes that are rich in fascin and somewhat stellate in morphology. These are the most helpful clues for distinguishing them from stained tumor cells (Figure 1).

    Either endothelial or dendritic cells in the samples were used as internal positive controls [4]. Nonimmune serum was used as the negative control.

    The differences between groups were investigated for statistical significance by the chi-square test, with p<0.05 as the cut-off point.
  • Top
  • Summary
  • Introduction
  • Materials And Methods
  • Results
  • Discussion
  • References
  • Results
    Fascin expression was noted in 17% (5/30) of the cases. All of the fascin-expressing GISTs were in the high-risk group (5/16), while the remaining high-risk as well as lowand moderate-risk GISTs showed no convincing immunoreactivity. The degree of staining was either moderate or strong (Figure 2). According to the basic four-level risk groups (Table 1) and eight-level groups based on the rates of metastases or tumor-related death (Table 2), the results seemed not statistically significant (p=0.261 and p=0.242, respectively).

    Fascin expression in GISTs composed of purely spindle cells was 4.5% (1/22) (Table 3). On the other hand, expression in the tumors composed of both spindle and epithelioid cells was around 50% (4/8) (Table 3). We observed that the intensity of staining was especially more conspicuous in epithelioid cells when compared to spindle cells. This finding seemed statistically significant (p=0.003). We also noted that the epithelioid cells, when present, were almost always positively labelled by fascin antibody (Figure 3).

    Fascin expression rates with respect to the localization of individual tumors are presented in Tables 1 and 2. No fascin expression was detected in GISTs from the large bowel (rectum) (0/3). We noticed a tendency of higher level of expression in the small bowel (jejunum and duodenum)-derived GISTs than those from the other localizations, and the difference was also statistically significant (p=0.043).

    All the five fascin-positive tumors were demonstrated to have some extent of infiltration into the nearby soft tissues and/or organs. One of the jejunal cases showed recurrence just six months after the primary surgical removal of the tumor.

    While epithelioid tumor cells were mostly positive for SMA as well, no immunoreactivity in spindle cells was detected. All fascin-positive cases were found negative for MSA.
  • Top
  • Summary
  • Introduction
  • Materials And Methods
  • Results
  • Discussion
  • References
  • Discussion
    Most of the gastrointestinal mesenchymal tumors, previously diagnosed as smooth muscle tumors, are now classified as GISTs. GISTs have a potential to show a wide variety of phenotypic differentiation including myoid, neural, mixed, and null morphology. Therefore, a broad immunohistochemical panel including muscular and neural markers such as CD 117 (c-kit), CD 34, SMA, desmin, and S-100 protein are required in their differential diagnosis. GISTs are usually positive for CD 117 (c-kit) and negative for S-100 and desmin (3,57,13). A small percentage of CD 117-negative GISTs are known to be positive for platelet-derived growth factor receptor-alpha (PDGFR-A), a receptor tyrosine kinase similar to CD 117 (5-7). Approximately 60-99% of all GISTs are also positive for CD 34, which is a progenitor-cell antigen mainly expressed by endothelial cells and in some subsets of fibroblasts and their tumors (6,7,13). About 10-30% of GISTs show a malignant clinical course (6). Pathologically, tumor location, size and mitotic activity are independent prognostic factors for GISTs (3,5).

    Fascin is an actin-bundling protein that is expressed in normal mesenchymal, endothelial, dendritic, and neuronal cells but not in normal epithelia (16). Many types of transformed epithelial cell lines and several epithelial tumors have been shown to express fascin (20-30). Some studies have suggested that the degree of fascin expression is well correlated with tumor stage, grade, lymph node involvement, and recurrence rate (22,23,29). However, this kind of relationship of fascin expression in spindle cell tumors has not yet been very well evaluated or documented. Grogg et al. (31) suggested that fascin is a potentially important marker for dendritic cell neoplasms, but it does not seem to be a useful marker in the diagnosis of other spindle cell tumors. In this study, we investigated the fascin expression in GISTs against their anatomic locations, invasiveness, prognostic risk groups, and the predominant morphology of neoplastic cells, either epithelioid, spindle or mixed.

    Grogg et al. (31) also investigated fascin expression in GISTs, and three cases (3/13 or 23%) showed only focal staining, while only one case (1/13 or 8%) showed diffuse expression. Similarly, we found that fascin expression is low in GISTs, in general (17%). Our results according to basic risk groups (Table 1) and groups based on tumor progression (Table 2) suggested that fascin expression in GISTs did not show any statistically significant differences (p=0.261 and p=0.242, respectively). However, fascin expressions with regard to tumor cell types and localizations seemed statistically significant (p=0.003 and p=0.043, respectively). We also noticed focal or individually fascin-labelled cells in some GISTs. We think that such cells, which represent only a small percentage (generally below 10%) of the entire tumor cells and show relatively weak staining (1+) compared to the intensely stained neoplastic cells, might be dendritic cells entrapped by the neoplastic growth. Our observation is supported by the study of Cameron et al. (32). They also demonstrated that dendritic cells were scattered between the GIST cells and were fascin-positive. In that case, the focally stained cells in GISTs, which were mentioned in the paper by Grogg et al. (31), like in our cases, should also be dendritic cells. Thus, we selectively ignored their positive staining and used them instead as positive internal control, among the positivity in endothelial cells.

    Smooth muscle actin (SMA) is reported positive in 25% of GISTs (8). Seidal et al. (9) found that MSA was weakly positive in 17% (11/64 cases) in GISTs. Because of its actin-bundling nature, we limited the actin immunoprofiling using SMA and MSA in only fascin-positive cases. There was no staining for MSA in our cases. While epithelioid cells showed SMA expression, there was no or only focal and weak staining in spindle cells. SMA expression was determined in fascin-positive cells and especially with epithelioid morphology. This difference might originate from different epitope or isomers of actins.

    In summary, our results indicated that the fascin expression was mostly moderate and limited only in high-risk GISTs. The other risk groups of GISTs showed practically no immunoreactivity with fascin. Staining intensity of epithelioid cells was significantly higher than that of the spindle cells. We noticed a tendency of higher level of expression in the small bowel (duodenum and jejunum) GISTs than those from the other localizations. These findings suggest that fascin expression in GISTs might be a well-correlated marker with the risk groups that are related to the tumor progression and aggressiveness.
  • Top
  • Summary
  • Introduction
  • Materials And Methods
  • Results
  • Discussion
  • References
  • References
    1. Appelman HD. Smooth muscle tumors of the gastrointestinal tract. What we know that Stout didn’t know. Am J Surg Pathol 1986; 10: 83-99.
    2. Bearzi I, Mandolesi A, Arduini F, et al. Gastrointestinal stromal tumor. A study of 158 cases: clinicopathological features and prognostic factors. Anal Quant Cytol Histol 2006; 28: 137-47.
    3. Hornick JL, Fletcher CD. The role of KIT in the management of patients with gastrointestinal stromal tumors. Hum Pathol 2007; 38: 679-87.
    4. Kindblom LG, Remotti HE, Aldenborg F, Meis-Kindblom JM. Gastrointestinal pacemaker cell tumor (GIPACT); gastrointestinal stromal tumors show phenotypic characteristics of the interstitial cells of Cajal. Am J Pathol 1998; 152: 1259-69.
    5. Miettinen M, Sobin LH, Lasota J. Gastrointestinal stromal tumors of the stomach: a clinicopathologic, immunohistochemical, and molecular genetic study of 1765 cases with long-term follow-up. Am J Surg Pathol 2005; 29: 52-68.
    6. Miettinen M, Makhlouf H, Sobin LH, et al. Gastrointestinal stromal tumors of the jejunum and ileum: a clinicopathologic, immunohistochemical, and molecular genetic study of 906 cases before imatinib with long-term followup. Am J Surg Pathol 2006; 30: 477-89.
    7. Miettinen M, Lasota J. Gastrointestinal stromal tumors: pathology and prognosis at different sites. Semin Diagn Pathol 2006; 23: 70-83.
    8. Miettinen M, Lasota J. Gastrointestinal stromal tumors: review on morphology, molecular pathology, prognosis, and differential diagnosis. Arch Pathol Lab Med 2006; 130: 1466-78.
    9. Seidal T, Edvardsson H. Expression of c-kit (CD117) and Ki67 provides information about the possible cell of origin and clinical course of gastrointestinal stromal tumours. Histopathology 1999; 34: 416-24.
    10. Ozcan A, Karslioglu Y, Kurt B, et al. Quantitative evaluation of immunohistochemical staining in gastrointestinal stromal tumors. Anal Quant Cytol Histol 2007; 29: 159-65.
    11. Tran T, Davila J, El-Serag HB. The epidemiology of malignant gastrointestinal stromal tumors: an analysis of 1,458 cases from 1992 to 2000. Am J Gastroenterol 2005; 100: 162-8.
    12. Tzen CY, Mau BL. Analysis of CD 117-negative gastrointestinal stromal tumors. World J Gastroenterol 2005; 11: 1052-5.
    13. Yamaguchi U, Hasegawa T, Masuda T, et al. Differential diagnosis of gastrointestinal stromal tumor and other spindle cell tumors in the gastrointestinal tract based on immunohistochemical analysis. Virchows Arch 2004; 445: 142-50.
    14. Robinson TL, Sircar K, Hewlett BR, et al. Short communication; gastrointestinal stromal tumors may originate from a subset of CD34 positive interstitial cells of Cajal. Am J Pathol 2000; 156: 1157-63.
    15. Tao YS, Edwards RA, Tubb B, et al. Beta-catenin associates with the actin-bundling protein fascin in a noncadherin complex. J Cell Biol 1996; 134: 1271-81.
    16. Kureishy N, Sapountzi V, Prag S, et al. Fascins, and their roles in cell structure and function. Bioessays 2002; 24: 350-61.
    17. Bryan J, Edwards R, Matsudaira P, et al. Fascin, an echinoid actin-bundling protein, is a homolog of the Drosophila singed gene product. Proc Nati Acad Sci 1993; 90: 9115-9.
    18. Adams JC. Roles of fascin in cell adhesion and motility. Curr Opin Cell Biol 2004; 16: 590-6.
    19. Hashimoto Y, Skacel M, Adams JC. Roles of fascin in human carcinoma motility and signaling: prospects for a novel biomarker? Int J Biochem Cell Biol 2005; 37: 1787-804.
    20. Goncharuk VN, Ross JS, Carlson JA. Actin-binding protein fascin expression in skin neoplasia. J Cutan Pathol 2002; 29: 430-8.
    21. Buda A, Pignatelli M. Cytoskeletal network in colon cancer: from genes to clinical application. Int J Biochem Cell Biol 2004; 36: 759-65.
    22. Hashimoto Y, Shimada Y, Kawamura J, et al. The prognostic relevance of fascin expression in human gastric carcinoma. Oncology 2004; 67: 262-70.
    23. Hashimoto Y, Ito T, Inoue H, et al. Prognostic significance of fascin overexpression in human esophageal squamous cell carcinoma. Clin Cancer Res 2005; 11: 2597-605.
    24. Hashimoto Y, Skacel M, Lavery IC, et al. Prognostic significance of fascin expression in advanced colorectal cancer: an immunohistochemical study of colorectal adenomas and adenocarcinomas. BMC Cancer 2006; 6: 241.
    25. Grothey A, Hashizume R, Sahin AA, McCrea PD. Fascin, an actin-bundling protein associated with cell motility, is upregulated in hormone receptor negative breast cancer. Br J Cancer 2000; 83: 870-3.
    26. Hu W, McCrea PD, Deavers M, et al. Increased expression of fascin, motility associated protein, in cell cultures derived from ovarian cancer and in borderline and carcinomatous ovarian tumors. Clin Exp Metastasis 2000; 18: 83-8.
    27. Jawhari AU, Buda A, Jenkins M, et al. Fascin, an actinbundling protein, modulates colonic epithelial cell invasiveness and differentiation in vitro. Am J Pathol 2003; 162: 69-80.
    28. Maitra A, Iacobuzio-Donahue C, Rahman A, et al. Immunohistochemical validation of a novel epithelial and a novel stromal marker of pancreatic ductal adenocarcinoma identified by global expression microarrays: sea urchin fascin homolog and heat shock protein 47. Am J Clin Pathol 2002; 118: 52-9.
    29. Pelosi G, Pastorino U, Pasini F, et al. Independent prognostic value of fascin immunoreactivity in stage I nonsmall cell lung cancer. Br J Cancer 2003; 88: 537-47.
    30. Puppa G, Maisonneuve P, Sonzogni A, et al. Independent prognostic value of fascin immunoreactivity in stage III–IV colonic adenocarcinoma. Br J Cancer 2007; 96: 1118-26.
    31. Grogg KL, Macon WR, Kurtin PJ, Nascimento AG. A survey of clusterin and fascin expression in sarcomas and spindle cell neoplasms: strong clusterin immunostaining is highly specific for follicular dendritic cell tumor. Mod Pathol 2005; 18: 260-6.
    32. Cameron S, Haller F, Dudas J, et al. Immune cells in primary gastrointestinal stromal tumors. Eur J Gastroenterol Hepatol 2008; 20: 327-34.
  • Top
  • Summary
  • Introduction
  • Materials And Methods
  • Results
  • Discussion
  • References
  • [ Summary ] [ PDF ] [ Similar Articles ] [ Mail to Author ] [ Mail to Editor ]
    Editorial Board | Instructions for Authors | Main Page | Table of Contents | Archive | Search