Stromal Reaction to Neoplasia: Colonic Carcinomas

The stroma and stromal reaction in the normal colon and in 14 different colonic tumors were studied by electron microscopy. Elastosis is a significant part of the stromal reaction to colonic adenocarcinomas and rectal squamous cell carcinomas. Two carcinoid tumors elicited no significant elastosis. In some of the adenocarcinomas, small muscular arteries close to neoplastic tissue developed massive elastosis of the media. This may indicate that the elastosis is due to stimulation of nonneoplastic stromal cells by some unknown neoplastic factor or factors.     

Stromal elastosis in papillary thyroid carcinomas

Stromal elastosis, defined as dense aggregations of elastic fibers, is found in some neoplastic tissues especially in malignant tumors of the breast and lung. Although also found in thyroid tissue, stromal elastosis in thyroid neoplasms have received little attention. To clarify the histopathological significance of stromal elastosis in the thyroid, we examined neoplastic (n = 223) and hyperplastic (n = 82) thyroid tissues in conjunction with cancer tissues (n = 193) of various other organs. Stromal elastosis was observed as deposits of pale homogeneous material in hematoxylin and eosin stain, and distinctively highlighted by elastic-van Gieson's stain. On immunohistochemical examination, elastin and tropoelastin were confirmed in these deposits. Stromal elastosis was found in 66% of papillary thyroid carcinomas (PTCs), although it was not identified in other histological types of thyroid neoplasms. In PTCs, deposits of elastic fibers varied in size and shape, and were more frequently distributed in the periphery of the tumor tissue. The histological subtypes of PTC varied in prevalence of elastosis with the follicular variant's (9%) prevalence being significantly lower than that of the classical type (72%). The frequency of stromal elastosis in PTCs was very similar to the frequencies in breast and lung adenocarcinomas, and higher than the frequencies in carcinomas of other organs. In conclusion, our results suggest that stromal elastosis is a characteristic histological finding of PTCs, presumably associated with their growth pattern and/or histological architecture. It is, therefore, reasonable to propose that stromal elastosis is an ancillary feature in the histopathological diagnosis of PTCs.     

Versican and hyaluronan expression in canine colonic adenomas and carcinomas: relation to malignancy and depth of tumour invasion

Changes in the production and structure of glycosaminoglycans and proteoglycans have been reported in many neoplastic tissues, and versican and hyaluronan (extracellular matrix components) are frequently increased in tumours and promote tumour progression. The distribution of chondroitin sulphate, versican and hyaluronan in normal canine colonic wall (n=10), and normal colonic lymph nodes (n=10), colonic adenomas (n=22), colonic adenocarcinomas (n=28), colonic undifferentiated carcinomas (n=7), and colonic lymph node metastases (n=8), was examined, with antibodies against chondroitin sulphate and versican, and a specific biotinylated probe for hyaluronan. The epithelial cells of the normal colonic mucosa were negative for all three substances, whereas the stromal tissue and lamina propria were moderately positive for chondroitin sulphate and hyaluronan, and weakly positive for versican. Chondroitin sulphate expression was increased in adenomas and carcinomas. However, there was no significant correlation between grade of tumour and degree of chondroitin sulphate expression. Versican expression was increased in the peritumoral stroma of adenocarcinomas and reduced in adenomas. A significant correlation was observed between grade of tumour and degree of versican expression. In 13 adenocarcinomas and undifferentiated carcinomas with invasion into all layers of the colorectum, the intensity of stromal versican expression was significantly related to the depth of invasion; the intensity was increased in the stroma of tumour islands in deep layers of the colonic wall. Unlike versican expression, hyaluronan expression was increased in the stromal tissue of both adenomas and carcinomas. However, the degree of stromal hyaluronan expression was unrelated to tumour grade and depth of tumour invasion. Hyaluronan was also expressed in the membrane and in the cytoplasm of tumour cells in 3/22 (14%) adenomas, 18/28 (64%) adenocarcinomas and 2/7 (29%) undifferentiated carcinomas. These results suggest that altered levels of both versican and hyaluronan in canine colonic tumours affect tumour progression.     

Comparative distribution of fibronectin and vitronectin in human breast and colon carcinomas. An immunofluorescence study

Distribution of two extracellular matrix components, vitronectin and fibronectin, was studied by immunofluorescence in 40 breast infiltrating duct carcinomas and in 15 colonic adenocarcinomas. Blood vessels, especially in peritumoral colonic submucosa, gave markedly different images after staining by antivitronectin antibody and antifibronectin serum. The former exclusively decorated the elastic laminae, whereas the latter gave a reaction much closer to the endothelium, probably localized at the basement membrane of the endothelium. Strong labeling of colonic and mammary stroma carcinomas by polyclonal antiserum against fibronectin was observed, whereas monoclonal antibody against vitronectin gave much more restricted labeling in the stroma of these tumors. In the stroma of colonic carcinomas, vitronectin was often situated in areas in which elastic fibers were visualized by orcein counterstaining. In breast stroma carcinomas it was always localized only in elastosis areas. Thus, it is likely that antivitronectin binds with glycoprotein(s) associated with elastic fibers. However, vitronectin staining also was seen in areas in which no elastic fibers were characterized by the orcein method.     

Duct elastosis in infiltrating carcinoma of the breast

Duct elastosis was studied in 219 patients subjected to radical mastectomy for infiltrating carcinoma of the breast, with a 10-year follow-up. Duct elastosis is a frequent finding in infiltrating breast cancer (65% of our cases). It develops in tumors of all three grades of malignancy, but it is more frequent in tumors of low grade malignancy (76% and 74% in grades I and II, respectively, and 47% in grade III tumors). In spite of their greater incidence in low malignancy tumors, the elastotic cases have a greater metastatic ratio than the non-elastotic cases (66% vs 45%). The elastotic cases also contain a significantly greater proportion of scirrhous tumors than the non-elastotic cases (86% vs. 32%). Duct elastosis and scirrhous reaction are two processes which develop in parallel, but are not related etiologically. They seem to be correlated with more advanced stages of the neoplastic disease. The influence of duct elastosis upon the ten year survival of the patients is unfavorable. this influence is not direct, and it is particularly evident in the metastatic cases. It seems to be related to the greater duration of the neoplastic disease and to the slow clinical course of tumors of low degree of malignancy.     

Elastosis in the male breast

Elastosis, a common finding in infiltrating ductal and lobular carcinomas of the female breast, has also been described in a variety of other benign and malignant lesions. However, elastosis has not been previously documented in male breast lesions. Ten cases of gynaecomastia and five neoplastic lesions of the male breast were evaluated for elastosis. In gynaecomastia, elastosis is a feature of the ducts surrounded by collagenized fibrous tissue but not of ducts with loose connective tissue stroma. This association is also evident to some extent in neoplastic lesions. These findings suggest that elastosis is an expression of periductal fibrosis irrespective of the nature of the lesion.     

Histologic parameters and DNA ploidy as predictors of survival in stage B adenocarcinoma of colon and rectum

Stage B adenocarcinomas of colon (61 carcinomas) and rectum (44 carcinomas) were retrospectively reviewed in order to define prognostic indicators and to determine if they are different for tumors arising at the two sites. Parameters evaluated included the substage of the tumor, histologic grade, presence of vascular/lymphatic invasion, residual adenoma or extracellular mucin, and tumor cell DNA content as determined by flow cytometry of paraffin-embedded tissues. Five-year survival was 81.0% for patients with colonic and 87.5% for those with rectal tumors. DNA ploidy was a significant predictor of overall and disease-free survival in patients with rectal adenocarcinomas (improved survival for those with diploid tumors) but not in those with colonic tumors and not in the two groups combined. The degree of differentiation of the tumor was prognostically significant in the colonic group and for the groups combined but not in the rectal group. There was a trend toward better survival in substage B1 tumors, as compared with substages B2 and B3, particularly in rectal neoplasms. Other histologic parameters did not predict survival. We conclude that the prognostic indicators in colonic adenocarcinoma seem to differ from those of adenocarcinoma arising in the rectum and that DNA ploidy has a significant impact upon outcome in stage B rectal adenocarcinomas.     

Monoclonal antibodies to cytoskeletal proteins: An immunohistochemical investigation of human colon cancer

Monoclonal antibodies raised to a number of microfilament-associated proteins were shown to recognize the appropriate proteins in extracts from human colon tissue. They were then used in an immunohistochemical study of normal colonic mucosa, adenomas, and adenocarcinomas. A strong reaction was seen in stromal cells within the tumours (both adenomas and adenocarcinomas) when frozen sections were stained with antibodies to filamin and caldesmon. In addition, a similar reaction was seen in the adenocarcinomas when stained with antibodies to talin and gelsolin. We believe that immunohistochemical staining with these antibodies reveals a tumour-induced process in the surrounding cells, possibly related to a host response to tumours.     

Laminin production by human endometrial stromal cells relates to the cyclic and pathologic state of the endometrium.

The cyclic changes in the presence of the basement membrane glycoprotein laminin in endometrial stromal cells was studied by immunohistochemistry. The interstitial matrix around the stromal cells of the proliferative phase of the normal menstrual cycle was unreactive with antibodies to laminin. However, commencing with the secretory phase, stromal cells accumulated distinct cytoplasmic and pericellular laminin-immunoreactive material. The maximal amount of stromal cell-associated laminin was observed in predecidual cells of the late secretory phase. Thus, laminin immunostaining discriminates stromal cells of the proliferative phase (being "negative") from those in the secretory phase (being "positive"). Sixty-six cases of endometrial hyperplasia and adenocarcinomas were also stained with antibodies to laminin. Sixty-nine percent of biopsies of cystic hyperplasia and 30% of adenomatous hyperplasia contained laminin-positive stromal cells. Ultrastructural examination of stromal cells in cystic hyperplasia revealed the presence of pericellular basement membrane-like material, focally arranged into typical lamina rara and lamina densa. In contrast, stromal cells in the atypical adenomatous hyperplasia and adenocarcinomas did not react with antibody to laminin. The expression of laminin receptor in the stromal cells codistributed with laminin. Basement membranes of the surface epithelium, the glandular epithelium, and the vessels stained strongly with antibodies to laminin. In preneoplastic and neoplastic tissues, laminin immunostaining revealed discontinuous and defective basement membranes. In poorly differentiated carcinomas only sparse amounts of laminin-positive basement membrane were observed; these tumors, in contrast, exhibited cytoplasmic laminin and also significant immunoreaction with antibodies to laminin receptor.     

The biological relevance of laminin 5gamma2 expression at the invading edge of colonic carcinomas

Previous studies have shown the presence of dilated neoplastic glands with cellular gaps called glandular pores (GPs) and laminin 5gamma2 expression at the invading edge of colonic carcinomas. We now extended our studies to explore a possible association between GP formation and laminin 5gamma2 expression at the invading edge of colonic carcinomas. Immunostain was performed on sections of five consecutive neoplastic glands with and without GPs from 86 colonic adenocarcinomas to assess the expression of laminin 5gamma2. Neoplastic glands with GPs were observed in 85% (73/86) of the tumors. Laminin 5gamma2 was expressed in 92% (335/365) of the neoplastic glands with GPs but only in 17% (63/365) of the neoplastic glands without GPs (p<0.05). Laminin 5gamma2 was overexpressed in the cells at the free ends of the pores in 88% of the neoplastic glands with GPs, but only in 14% of those without pores (p<0.05). Hence, at the growing edge of colonic carcinomas, laminin 5gamma2 was frequently expressed in neoplastic glands having GPs. Remarkably, the tumor cells at the free ends of the GPs overexpressed laminin 5gamma2, indicating increased production of this adhesion-migration macromolecule. The results suggest a close interaction between this adhesion-migration macromolecule, PG formation and the local progression of colonic carcinomas.     

Coexistent adenomyoepithelioma and invasive ductal carcinoma of the breast: presentation as separate tumors

Adenomyoepitheliomas are rare breast tumors. We report an unusual case of adenomyoepithelioma associated with invasive ductal carcinoma here. Histologically, the lesion consisted of two separate tumors. One nodule corresponded to invasive ductal carcinoma consisting of tubular and trabecular arrangements of columnar or cuboidal neoplastic cells. The other tumor corresponded to adenomyoepithelioma consisting of an inner layer of neoplastic cells with basophilic cytoplasm and the outer layer of neoplastic cells with clear cytoplasm. Immunohistochemically, some myofibroblasts were observed in the stroma of both adenomyoepithelioma and invasive ductal carcinoma, but no CD34-positive stromal cells were seen in the stroma of either lesion. The stromal reaction of adenomyoepithelioma resembles that of intraductal papilloma in the previous study. To the best of our knowledge, this is the first case of coexistent adenomyoepithelioma and invasive ductal carcinoma of the breast that were discovered as separate nodules. Clinicians and pathologists should be aware of such an association because they need to distinguish such a case from malignant neoplasms arising in adenomyoepithelioma. Additionally, our preliminary report suggests that the stromal response of adenomyoepithelioma may resemble that of intraductal papilloma.     

Platelet-derived growth factor causes pulmonary cell proliferation and collagen deposition in vivo.

Malignant ovarian tumors induce a strong fibro-proliferative reaction characterized by the active production of type I and type III procollagen both locally in the ovary as well as more remotely in the peritoneal cavity. Our purpose was to determine the origin of the increased collagen production observed in serous ovarian tumors with different histological grades of malignancy, ie, whether the malignant cells or the stromal fibroblasts are responsible for the synthesis of collagen fibers. We visualized the mRNAs corresponding to the pro alpha 1(I) and pro alpha 2(I) chains of type I procollagen and the pro alpha 1(III) chain of type III procollagen by in situ hybridization. Strong signals for both chains of type I procollagen were seen in stromal fibroblasts next to tumor cell islets, whereas the reaction was weak or absent near benign ovarian cysts. In poorly differentiated tumors, the signals were particularly abundant and occasionally also seen in the neoplastic cells themselves. Type III procollagen mRNA expression was similar, although somewhat less distinct. These findings indicate that the production of interstitial procollagens is related to the degree of malignancy and neoplastic activity of tumors. The formation of collagen in well differentiated ovarian tumors is a function of stromal fibroblasts, whereas in poorly differentiated tumors, aberrant expression of one or several chains of type I and type III procollagens in the neoplastic cells is also likely to take place.     

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

We extended the results of a previous microarray analysis by immunohistochemical validation of differential protein expression in a series of 57 surgically resected infiltrating ductal pancreatic adenocarcinomas. Two representative genes were examined: sea urchin fascin homolog (overexpressed in both cell lines and primary tumors) and heat shock protein 47 (HSP47; overexpressed in primary tumors only). Protein expression also was evaluated in the precursor lesions of pancreatic cancer pancreatic intraepithelial neoplasia (PanIN), and normal ductal epithelium. Fascin expression was seen in the neoplastic cells of 54 (95%) of 57 ductal adenocarcinomas but not in 49 (94%) of 52 adjacent nonneoplastic epithelium. In the multistep pathogenesis of ductal adenocarcinomas, fascin expression seemed to be a late event, usually present in PanINs 2 and 3. HSP47 expression was almost universal and most intense in the ductal adenocarcinoma-associated stromal desmoplasia (57/57), although 37 cases (65%) also expressed HSP47 in the neoplastic epithelium. HSP47 expression was absent in the majority of nonneoplastic pancreata (46 [88%]). Fascin and HSP47 are novel tumor markers with potential diagnostic and therapeutic implications for pancreatic carcinoma. These results establish the usefulness of global expression platforms to identify novel tumor markers.     

Genetic model of multi-step breast carcinogenesis involving the epithelium and stroma: clues to tumour-microenvironment interactions

Although numerous studies have reported that high frequencies of loss of heterozygosity (LOH) at various chromosomal arms have been identified in breast cancer, differential LOH in the neoplastic epithelial and surrounding stromal compartments has not been well examined. Using laser capture microdissection, which enables separation of neoplastic epithelium from surrounding stroma, we microdissected each compartment of 41 sporadic invasive adenocarcinomas of the breast. Frequent LOH was identified in both neoplastic epithelial and/or stromal compartments, ranging from 25 to 69% in the neoplastic epithelial cells, and from 17 to 61% in the surrounding stromal cells, respectively. The great majority of markers showed a higher frequency of LOH in the neoplastic epithelial compartment than in the stroma, suggesting that LOH in neoplastic epithelial cells might precede LOH in surrounding stromal cells. Furthermore, we sought to examine pair-wise associations of particular genetic alterations in either epithelial or stromal compartments. Seventeen pairs of markers showed statistically significant associations. We also propose a genetic model of multi-step carcinogenesis for the breast involving the epithelial and stromal compartments and note that genetic alterations occur in the epithelial compartments as the earlier steps followed by LOH in the stromal compartments. Our study strongly suggests that interactions between breast epithelial and stromal compartments might play a critical role in breast carcinogenesis and several genetic alterations in both epithelial and stromal compartments are required for breast tumour growth and progression.     

Lysozyme and alpha 1-antitrypsin in giant-cell tumor of bone and in other lesions that contain giant cells

We performed an immunohistochemical study of 24 giant-cell tumors of bone and 30 other lesions (fibrous histiocytoma, nonossifying fibroma, and giant-cell tumor of the tendon sheath) using lysozyme and alpha 1-antitrypsin as markers for histiocytic cells. The presence of histiocytic cells in giant-cell tumors of bone is confirmed by the finding of a positive reaction for alpha 1-antitrypsin in both multinucleate giant cells and mononuclear stromal cells in some cases. It is not clear whether the positive cells are to be regarded as neoplastic or reactive and alpha 1-antitrypsin is not considered as a diagnostically useful marker for giant-cell tumor of bone. In malignant fibrous histiocytoma, too, histiocytic cells could be identified by their positive reaction for alpha 1-antitrypsin; some of these cells had the morphologic features of tumor cells. Cells with a positive reaction for lysozyme were rarely found, except in giant-cell tumors of the tendon sheath.     

CD34+ fibrocytes in neoplastic and inflammatory pancreatic lesions

Besides its function as a matrix-producing cell, the CD34+ fibrocyte has been reported to be an antigen-presenting cell capable of priming naive T cells in situ. Therefore, it has been claimed that the CD34+ fibrocyte may play an important role in host response to tissue damage. The objective of the present study was to analyze the presence and distribution of CD34+ fibrocytes and smooth muscle actin (SMA) reactive myofibroblasts in relation to the underlying pancreatic disease. We investigated a total of 12 pancreatic adenocarcinomas, 7 endocrine tumors of the pancreas, and 8 cases of chronic pancreatitis; in 11 cases, normal pancreatic tissue was available. The stroma of normal pancreatic tissue harbored diffusely scattered CD34+ fibrocytes. Chronic pancreatitis was characterized by an increased number of stromal CD34+ fibrocytes paralleled by a gain of SMA reactive myofibroblasts which were not observed in the normal pancreatic stroma. The stroma of pancreatic ductal adenocarcinomas and endocrine tumors was devoid of CD34+ fibrocytes or showed at least a focal loss of this cell type, whereas SMA reactive myofibroblasts were detected in both endocrine tumors and adenocarcinomas. We conclude that detection of CD34+ fibrocytes may constitute an adjunctive tool in distinguishing chronic pancreatitis from ductal adenocarcinoma since the absence of this cell population strongly favors a neoplastic process. Moreover, CD34+ fibrocytes and myofibroblasts appear to be involved in stromal remodeling associated with chronic pancreatitis and ductal adenocarcinoma.     

E-cadherin nuclear staining is useful for the diagnosis of ovarian adult granulosa cell tumor

We recently have demonstrated nuclear localization of E-cadherin in ovarian adult granulosa cell tumors (Histopathology 2011;58:423). The purpose of the present study is to investigate the diagnostic utility of E-cadherin nuclear staining for the differential diagnosis between ovarian adult granulosa cell tumor and its morphological mimics. Tissue samples taken from 81 ovarian tumors and 20 extraovarian tumors were immunohistochemically stained using monoclonal anti-E-cadherin antibody recognizing cytoplasmic domain (clone 36 supplied by BD Biosciences, San Jose, CA). The ovarian tumors consisted of 30 adult granulosa cell tumors, 3 Sertoli-stromal cell tumors, 14 fibrothecomas, 5 carcinoid tumors, 1 large cell neuroendocrine carcinoma, 18 endometrioid adenocarcinomas, and 10 poorly differentiated serous adenocarcinomas. Extraovarian tumors consisted of 16 uterine endometrial stromal neoplasms and 4 pulmonary small cell carcinomas. Only tumor cells with nuclear staining were considered positive in this study. Ninety percent of adult granulosa cell tumors, 67% of Sertoli-stromal cell tumors, 64% of fibrothecomas, 75% of endometrial stromal neoplasms, 75% of small cell carcinomas, and the one large cell neuroendocrine carcinoma showed E-cadherin nuclear expression, whereas all the ovarian carcinoid tumors, endometrioid adenocarcinomas, and poorly differentiated serous adenocarcinomas were negative. E-cadherin nuclear staining is useful in distinguishing between adult granulosa cell tumors and ovarian adenocarcinomas or carcinoid tumors. However, it is of limited use for distinguishing between adult granulosa cell tumors and endometrial stromal neoplasms or small cell carcinomas. E-cadherin should be included in the immunohistochemical panel for an accurate diagnosis of ovarian adult granulosa cell tumors.     

Cutis rhomboidalis protects skin from malignant epithelial tumors

Cutis rhomboidalis nuchae is a skin alteration which comes from chronic sun exposure and it integrates the solar elastosis group, acquiring a coriaceous aspect, with a yellowish and grooved surface. There is the occurrence of elastic and collagen fibers degeneration found in the dermis caused by ultraviolet radiation [1]. Another group of skin diseases which has solar exposure as a determining factor is the group of actinic keratoses, the non-melanoma malignant epithelial tumors {basal cell carcinoma (CBC) and squamous cell carcinoma (CEC)} [2].However, the occurrence of actinic keratoses, CBCs or CECs on the area of cutis rhomboidalis is infrequent in dermatology clinical practice. The authors do not know why people with neoplasias and pre neoplastic lesions in some areas with chronic photo damage amendments (face and upper limbs), do not present the same pre and neoplastic lesions in areas with similar appearance of chronic sun damage (nape). The authors seek to understand why the nape is protected for pre and neoplastic lesions. We suggest that cutis rhomboidalis protects skin from malignant epithelial tumors in nuchae.