Filippo Fraggetta

Independent prognostic value of fascin immunoreactivity in stage I nonsmall cell lung cancer

Fascin-1, the most expressed form of fascin in vertebrate tissues, is an actin-bundling protein that induces cell membrane protrusions and increases motility of normal and transformed epithelial cells. Very few data are available on the role of this protein in nonsmall cell lung cancer (NSCLC). Two hundred and twenty patients with stage I NSCLC and long-term follow-up were evaluated immunocytochemically for fascin expression. Overall, variable fascin immunoreactivity was detected in 98% of 116 squamous cell carcinomas, in 78% of 96 adenocarcinomas, in 83% of six large cell carcinomas, and in the two adenosquamous carcinomas under study. Neoplastic emboli were commonly decorated by the antifascin antibody (P<0.001), also when the surrounding invasive carcinoma was unreactive. Fascin immunoreactivity correlated with high tumour grade (P=0.017) and, in adenocarcinomas, with high Ki-67 labelling index (P=0.021). Adenocarcinomas with a prevalent bronchiolo-alveolar in situ component were less commonly immunoreactive for fascin than invasive tumours (P=0.005). Contralateral thoracic or distant metastases were associated significantly with diffuse (>60% immunoreactive tumour cells) fascin expression in adenocarcinomas (P=0.043), and marginally with strong fascin immunostaining in squamous cell carcinomas (P=0.13). No associations were noted with any other clinicopathological variables tested. Patients with tumours showing diffuse (>60% immunoreactive neoplastic cells) and/or strong immunoreactivity for fascin had a shorter survival (P=0.006 for adenocarcinomas and P=0.026 for squamous cell carcinomas), even after multivariate analysis (P=0.014 and 0.050, respectively). The current study documents for the first time that fascin is upregulated in invasive and more aggressive NSCLC, being an independent prognostic predictor of unfavourable clinical course of the disease. Targetting the fascin pathway could be a novel therapeutic strategy of NSCLC.

p63 immunoreactivity in lung cancer: yet another player in the development of squamous cell carcinomas?

The p63 protein, a member of the p53 family of nuclear transcription factors, is characterized by different capabilities of transactivating reporter genes, inducing apoptosis, and functioning as dominant‐negative agent. This study evaluated the prevalence and prognostic implications of p63 immunoreactivity in 221 patients with stage I non‐small cell lung carcinoma (NSCLC) and in 57 patients with stage I–IV neuroendocrine tumours (NET). The results were correlated with the tumour proliferative fraction, the accumulation of p53 protein, and with patient survival. p63 immunoreactivity was seen in 109/118 squamous cell carcinomas, 15/95 adenocarcinomas, 2/2 adenosquamous carcinomas, 4/6 large cell carcinomas, 9/20 poorly differentiated NET, and 1/37 typical and atypical carcinoids (p < 0.001). Furthermore, the prevalence of p63‐immunoreactive cells increased progressively from pre‐neoplastic and pre‐invasive lesions to invasive squamous cell carcinomas. In these latter tumours, but not in adenocarcinomas, p63 immunoreactivity correlated directly with the tumour proliferative fraction (p = 0.028), and inversely with the tumour grade (p = 0.004). No relationship was found with p53 protein immunoreactivity or the other clinico‐pathological variables examined. Although p63 is likely to be involved in the development of pulmonary squamous cell carcinoma, it does not carry any prognostic implication for NSCLC patients. Copyright

Pleomorphic carcinomas of the lung show a selective distribution of gene products involved in cell differentiation, cell cycle control, tumor growth, and tumor cell motility: a clinicopathologic and immunohistochemical study of 31 cases.

We investigated 31 cases of pleomorphic carcinomas of the lung, with a double component of neoplastic epithelial cells and of spindle and/or giant cells. To correlate the morphologic diversity of these two cell components with their immunophenotype, we evaluated the expression of several gene products involved in cell differentiation (cytokeratins, epithelial membrane antigen, carcinoembryonic antigen, vimentin, S-100 protein, smooth muscle actin, desmin), cell cycle control and apoptosis (p53, p21Waf1, p27Kip1, FHIT), tumor growth (proliferative fraction, assessed by Ki-67 antigen, and microvascular density, assessed by CD34 immunostaining), and tumor cell motility (fascin). We found the epithelial component to be significantly more immunoreactive for cytokeratins, epithelial membrane antigen, carcinoembryonic antigen, cell cycle inhibitors p21Waf1, p27Kip1 and tumor suppressor gene FHIT, whereas the sarcomatoid component, independent of tumor stage and size, was more immunoreactive for vimentin, fascin, and microvascular density. Accordingly, we suggest a model of tumorigenesis whereby the mesenchymal phenotype of pleomorphic cells is likely induced by the selective activation and segregation of several molecules involved in cell differentiation, cell cycle control, and tumor cell growth and motility. Whether pleomorphic carcinomas of the lung are tumors with a dismal prognosis still remains an unsettled issue. In our series, however, stage I pleomorphic carcinomas have the same clinical behavior as ordinary non-small cell lung cancer, and only a high proliferative index (Ki-67 labeling index >35%) is associated with a worse prognosis in these tumors.

Independent value of fascin immunoreactivity for predicting lymph node metastases in typical and atypical pulmonary carcinoids

Immunoreactivity for fascin, an actin-bundling protein related to cell motility, has been reported in breast, ovary, pancreas, skin, and non-small cell carcinomas, and associated with more advanced disease stage and poorer prognosis. Data on pulmonary neuroendocrine (NE) tumors, however, are lacking. We evaluated the expression of fascin by immunohistochemistry--using two different monoclonal antibodies--in surgical specimens of pulmonary NE tumors of all the diverse histological types from 128 consecutive patients recruited between 1987 and 2001, and investigated its relationship with the presence of lymph node metastases. Overall, fascin immunoreactivity was detected in 5% of 38 typical carcinoids (TC), 35% of 23 atypical carcinoids (AC), 83% of 40 large-cell neuroendocrine carcinomas (LCNEC), and 100% of 27 small-cell lung carcinomas (SCLC) (P<0.001), Normal NE cells or hyperplastic NE tumorlets were consistently unreactive. No statistically significant differences in fascin immunoreactivity were found between the two antibodies. In TC and AC but not high-grade NE tumors, fascin immunoreactivity closely correlated with the occurrence of lymph node metastases, the pN class and the number of involved lymph nodes (P<0.001). It was also significantly associated with an increased proliferative activity (Ki-67 labeling index >5%) (P=0.020), and with either down-regulation or altered subcellular compartmentalization of E-cadherin (P<0.001) and CD99 (P=0.030), two cell adhesion complexes in pulmonary NE tumors. At multivariate analysis, only fascin emerged as an independent predictor of lymph node metastases in this tumor group (HR 30.28; 95% confidence intervals: 1.59-574.49; P=0.023). This study indicates that fascin immunoreactivity may identify subsets of pulmonary carcinoid patients with different metastatic potential to regional lymph nodes. Targeting the fascin pathway could be a novel therapeutic strategy of pulmonary carcinoids.

CDX2 immunoreactivity in primary and metastatic ovarian mucinous tumours

The caudal-related homeobox transcription factor CDX2 regulates the differentiation of intestinal epithelial cells. Recent studies have suggested that CDX-2 immunoreactivity is strictly confined to benign and malignant intestinal epithelium. In the present study, we evaluated the prevalence of CDX2 immunoreactivity in a series of benign, borderline and malignant primary ovarian mucinous neoplasms. We tested 62 mucinous tumours of the ovary, including 28 benign cystadenomas, 18 borderline tumours, 16 adenocarcinomas, 35 serous and endometrioid ovarian lesions and 10 ovarian metastases of colonic adenocarcinoma. Overall, the CDX2 prevalence in primary mucinous tumours was 79%, including 20 of 28 (71.5%) cystadenomas, 14 of 18 (77.7%) borderline tumours and 15 of 16 (93.5%) adenocarcinomas. Immunoreactivity usually correlated with intestinal differentiation of tumour cells, although wide heterogeneity in the distribution of immunolabelled cells was noted. No immunoreactivity was observed in serous lesions; whereas, 1 of 13 (7.7%) endometrioid adenocarcinomas and all of the 10 metastatic colonic adenocarcinomas were immunostained. These results indicate that CDX2 is detectable in the majority of benign, borderline and malignant ovarian mucinous tumours and, therefore, makes this marker unsuitable when distinguishing primary from metastatic ovarian mucinous adenocarcinomas. However, CDX2 immunoreactivity could be useful in the distinction between endocervical and intestinal-type mucinous tumours of the ovary, which may have clinical relevance.

PARP-1 protein expression in glioblastoma multiforme

One of the most common type of primary brain tumors in adults is the glioblastoma multiforme (GBM) (World Health Organization grade IV astrocytoma). It is the most common malignant and aggressive form of glioma and it is among the most lethal ones. Poly (ADP-ribose) polymerase 1 (PARP-1) gene, located to 1q42, plays an important role for the efficient maintenance of genome integrity. PARP-1 protein is required for the apoptosis-inducing factor (AIF) translocation from the mitochondria to the nucleus. PARP-1 is proteolytically cleaved at the onset of apoptosis by caspase-3. Microarray analysis of PARP-1 gene expression in more than 8000 samples revealed that PARP-1 is more highly expressed in several types of cancer compared with the equivalent normal tissues. Overall, the most differences in PARP-1 gene expression have been observed in breast, ovarian, endometrial, lung, and skin cancers, and non-Hodgkins lymphoma. We evaluated the expression of PARP-1 protein in normal brain tissues and primary GBM by immunohistochemistry. Positive nuclear PARP-1 staining was found in all samples with GBM, but not in normal neurons from controls (n=4) and GBM patients (n=27). No cytoplasmic staining was observed in any sample. In conclusion, PARP-1 gene is expressed in GBM. This finding may be envisioned as an attempt to trigger apoptosis in this tumor, as well as in many other malignancies. The presence of the protein exclusively at the nucleus further support the function played by this gene in genome integrity maintenance and apoptosis. Finally, PARP-1 staining may be used as GBM cell marker.

CD99 immunoreactivity in gastrointestinal and pulmonary neuroendocrine tumours

Abstract Although considered a specific marker for Ewing’s sarcoma/peripheral neuroectodermal tumour, the MIC2 gene product (CD99) has been immunolocalised in a variety of human tumours. The present study evaluated immunohistochemically the prevalence of CD99 expression in a series of 68 neuroendocrine tumours of different gastrointestinal and pulmonary sites. We now report on membrane and/or granular cytoplasmic immunoreactivity in 25% of these tumours, independent of their anatomical sites. In lung neuroendocrine tumours, CD99 was preferentially confined to typical carcinoids (P=0.009). A statistically significant relationship was observed between the number of CD99 positive cells but not the immunostaining patterns and the presence of local invasion and/or distant metastases (P<0.001). Moreover, there was a tendency for CD99-reactive tumours to show a reduced proliferative activity expressed by a Ki67 index of 2% (P=0.119). The number of CD99 immunoreactive cells or patterns of immunoreactivity did not correlate with the presence of associated clinical syndrome or particular hormonal immunostaining. Although the molecular basis underlying CD99 expression in neuroendocrine tumours is still poorly understood, our data suggest that CD99 may be involved in cell-to-cell adhesion of neuroendocrine tumour cells and in downregulation of their proliferative activity.

Myofibroblastoma of the Breast with Hemangiopericytoma-like Pattern and Pleomorphic Lipoma-like areas : Report of a Case with Diagnostic and Histogenetic Considerations

Myofibroblastoma (MFB) of the breast is an uncommon benign spindle cell tumor which may exhibit a wide spectrum of histological features. We report an unusual case of MFB of the male breast, showing cellular areas with a hemangiopericytoma-like pattern similar to that observed in solitary fibrous tumor (SFT) and extensive fibromyxoid areas containing numerous atypical stromal cells. The association of these atypical cells with mature adipocytes and microcystic and/or myxoid degenerative changes resembled pleomorphic lipoma-like and myxoid liposarcoma-like features, respectively. To our knowledge, these peculiar morphological findings have not been previously reported in MFB of the breast. They should be recognized to avoid confusion with other mesenchymal tumors, especially with hemangiopericytoma, pleomorphic lipoma (PL), spindle-cell lipoma (SCL) and myxoid liposarcoma. A case of MFB of the breast showing morphological features also commonly seen in SFT and PL/SCL is further morphological evidence in support of the speculation that the mesenchymal tumors of the breast, also known under the terms benign spindle cell tumors, fibromas, SFTs, SCLs and MFBs, are histogenetically related lesions.

Primitive neuroectodermal tumour of the uterus with focal cartilaginous differentiation

The uterus was enlarged (130 × 80 × 40 mm) with a smooth external surface. The endometrial cavity was filled by a white-grey, polypoidal, fleshy mass arising from the posterior wall, and measuring 60 mm in maximum dimension (Figure 1). Areas of necrosis and haemorrhage were present and, focally, the tumour invaded the myometrium but did not reach the peritoneal surface. The ovaries and Fallopian tubes were unremarkable: a paratubal cyst was present on the right. Histology revealed a neoplasm composed of solid sheets of undifferentiated round to oval cells forming numerous rosettes. The rosettes were mostly of the Flexner–Wintersteiner type (Figure 2a), but some Homer–Wright type and perivascular pseudorosettes were occasionally detected. In some areas, neuroectodermal columnar cells were arranged in medulloepithelial tubules dispersed through a loose fibrillary matrix (Figure 2b). The neoplastic cells had scanty cytoplasm, oval, hyperchromatic nuclei and inconspicuous nucleoli. Mitotic figures were absent or rare. Necrotic and haemorrhagic areas were scattered throughout the tumour. No lymphatic or vascular invasion was seen. Most of the endometrium was replaced by tumour and uninvolved endometrium was atrophic with occasional cystically dilated glands (Figure 2a). Despite extensive sampling, we found only a small island of mature cartilage which was confirmed immunohistochemically by S-100 protein positivity (Figure 2c). No other mesodermal or epithelial elements were identified within the neoplasm. At margin of the mass, tumour infiltrated the superficial myometrium. The Fallopian tubes and ovaries were atrophic, in keeping with the patient’s age. There was no evidence of metastatic tumour in the pelvic lymph nodes. The tumour was classified as stage IB (FIGO). Immunohistochemistry was carried out using standard avidin-biotin-peroxidase complex (ABC). The following antibodies were used: neuron-specific-enolase Histopathology 1997, 30, 483–485

Poly (ADP-ribose) Polymerase 1 Protein Expression in Normal and Neoplastic Prostatic Tissue

A genetic background has been implicated in the development of prostate cancer. Protein microarrays have enabled the identification of proteins, some of which associated with apoptosis, that may play a role in the development of such a tumor. Inhibition of apoptosis is a co-factor that contributes to the onset and progression of prostate cancer, though the molecular mechanisms are not entirely understood. Poly (ADP-ribose) polymerase 1 (PARP-1) gene is required for translocation of the apoptosis-inducing factor (AIF) from the mitochondria to the nucleus. Hence, it is involved in programmed cell death. Different PARP-1 gene expression has been observed in various tumors such as glioblastoma, lung, ovarian, endometrial, and skin cancers. We evaluated the expression of PARP-1 protein in prostatic cancer and normal prostate tissues by immunohistochemistry in 40 men with prostate cancer and in 37 normal men. Positive nuclear PARP-1 staining was found in all samples (normal prostate and prostate cancer tissues). No cytoplasmic staining was observed in any sample. PARP-1-positive cells resulted significantly higher in patients with prostate carcinoma compared with controls (P<0.001). PARP-1 over-expression in prostate cancer tissue compared with normal prostate suggests a greater activity of PARP-1 in these tumors. These findings suggest that PARP-1 expression in prostate cancer is an attempt to trigger apoptosis in this type of tumor similarly to what reported in other cancers.