Diagnosis of a peripheral neuroectodermal tumour by fine needle aspiration

One case of malignant peripheral neuroectodermal tumour successfully diagnosed by cytology is presented. Although a Papanicolaou stained smear could not lead to a diagnosis more specific than a malignant small cell tumour, ancillary analytic methods performed on the cytologic material including immunocytochemistry and electron microscopy yielded the correct diagnosis of peripheral neuroectodermal tumour. This case demonstrates that a precise categorization of small round cell tumours may be achieved by cytology as long as some material is kept for immunocytochemical and ultrastructural studies.     

Primitive Neuroectodermal Tumour (PNET) in subcutaneous abdominal wall: a case report

Primitive neuroectodermal tumour (PNET) is a rare tumour mainly found in children under ten years old. It may be broadly categorised into those occurring from the central or peripheral nervous system of which the majority arise centrally. We report a 61 year-old lady who had previous lobular breast cancer presenting with a rapidly expanding lesion in her anterior right upper abdominal wall. Clinically it appeared to be benign, however, histopathology of the excised lesion revealed a localised PNET. This case is an unusual case of a PNET in an adult that is peripheral in nature arising from subcutaneous tissue in the abdominal wall.     

Surgical treatment of neuroendocrine tumours of the pancreas

Gastro-entero-pancreatic (GEP) endocrine tumours can originate from various pancreatic islet cells, from endocrine cells of the gastric and duodenal mucosa, or from APUD cells of neuroectodermal origin in the gastrointestinal tract. They are benign when smaller than 2 cm, but larger tumours are generally malignant. Surgery is the only method for the curative treatment of GEP tumours. A diagnosed and localised tumour is an absolute indication for radical surgery. Conservative medical treatment may be indicated only in an inoperable condition, but in this case tumour reduction surgery is suggested. In the last 15 years 22 patients with pancreatic neuroendocrine tumours were treated without any mortality. Except for two of them, the surgical therapy was curative.     

Fate of amnion-derived stem cells transplanted to the fetal rat brain: migration, survival and differentiation

We have recently characterized a stem cell population isolated from the rodent amniotic membrane termed amnion-derived stem cells (ADSCs). In vitro ADSCs differentiate into cell types representing all three embryonic layers, including neural cells. In this study we evaluated the neuroectodermal potential of ADSCs in vivo after in utero transplantation into the developing rat brain. A clonal line of green fluorescent protein-expressing ADSCs were infused into the telencephalic ventricles of the developing embryonic day 15.5 rat brain. At E17.5 donor cells existed primarily as spheres in the ventricles with subsets fused to the ventricular walls, suggesting a mode of entry into the brain parenchyma. By E21.5 green fluorescent protein (GFP) ADSCs migrated to a number of brain regions. Examination at postnatal time points revealed that donor ADSCs expressed vimentin and nestin. Subsets of transplanted ADSCs attained neuronal morphologies, although there was no immunohistochemical evidence of neural or glial differentiation. Some donor cells migrated around blood vessels and differentiated into putative endothelial cells. Donor ADSCs transplanted in utero were present in recipients into adulthood with no evidence of immunological rejection or tumour formation. Long-term survival may suggest utility in the treatment of disorders where differentiation to a neural cell type is not required for clinical benefit.     

Increased unsaturation of lipids in cytoplasmic lipid droplets in DAOY cancer cells in response to cisplatin treatment

Increases in ¹H nuclear magnetic resonance spectroscopy (NMR) visible lipids are a well-documented sign of treatment response in cancers. Lipids in cytoplasmic lipid droplets (LDs) are the main contributors to the NMR lipid signals. Two human primitive neuroectodermal tumour cell lines with different sensitivities to cisplatin treatment were studied. Increases in NMR visible saturated and unsaturated lipids in cisplatin treated DAOY cells were associated with the accumulation of LDs prior to DNA fragmentation due to apoptosis. An increase in unsaturated fatty acids (UFAs) was detected in isolated LDs from DAOY cells, in contrast to a slight decrease in UFAs in lipid extracts from whole cells. Oleic acid and linoleic acid were identified as the accumulating UFAs in LDs by heteronuclear single quantum coherence spectroscopy (HSQC). ¹H NMR lipids in non-responding PFSK-1 cells were unchanged by exposure to 10 μM cisplatin. These findings support the potential of NMR detectable UFAs to serve as a non-invasive marker of tumour cell response to treatment.     

Phenotypic variations and dynamic topography of transformed cells in an experimental model of diethylstilbestrol-induced renal tumour in male Syrian hamster

This work explores the phenotypic changes affecting transformed cells in an experimental model of diethylstilbestrol (DES)-induced renal tumours in male Syrian hamster. This estrogen-induced neoplasm presents an important cytological pleomorphism and its origin remains largely controversial. In order to characterize phenotypic variations during tumour progression, the occurrence of seven lineage markers was analysed by a morphometric approach in kidney sections of DES-exposed hamsters (6–;11 months). S100 protein, neuron-specific enolase (NSE) and vimentin are expressed by a large percentage of malignant cells during tumour development. Glial fibrillary acidic protein (GFAP), protein gene product 9.5 (PGP 9.5) and desmin are mostly evidenced in advanced neoplasm whereas Leu 7 always presents a focal expression. As evidenced by double-label immunofluorescence, the coexpression of three important neuroectodermal lineage markers (S100, NSE and PGP 9.5) in earliest tumour buds points to a peripheral nerve sheath origin for this neoplasm thus confirming previously published data. For each marker, the fluctuations of expression levels during tumour progression as well as the spatial heterogeneity of distribution suggest variable phenotypic differentiation of transformed cell populations. This observation is largely corroborated by double-label immunofluorescence showing coexpression modification of several markers during tumour progression. This points to a complex dynamic and spatial self-organization of different phenotypes within neoplasms. Altogether, these results support the concept that DES-induced kidney tumours are not made of unstructured cell populations but represent adaptive complex dynamic biosystems.     

Primitive neuroectodermal tumor of the uterus. A case report

BACKGROUND: Primitive neuroectodermal tumor (PNET) is a rare tumor derived from fetal neuroectodermal cells. These tumors occur in the central nervous system and in peripheral locations. Histologic diagnosis is the standard since most of these tumors are detected at an advanced stage. CASE: A 17-year-old female presented with persistent vaginal bleeding. Physical examination revealed a 4-cm, hard, barrel-shaped cervix. A cervicovaginal smear was obtained. The specimen was hypercellular, with small to medium-sized, round, malignant cells. A diagnosis of PNET was made from the histologic sections of the surgical specimen. CONCLUSION: When numerous small round cells in a diffuse pattern are seen on a Pap smear, the differential diagnosis is long and difficult. However, with careful evaluation of the cytologic features, a few reasonable differential diagnoses can be reached. Furthermore, with liquid-based Pap smears, material is available for immunohistochemical staining to narrow the range even more. Using all resources, including a good clinical history, a cytopathologist can give the clinician an early diagnosis for intervention and treatment.     

Nestin, a neuroectodermal stem cell marker molecule, is expressed in Leydig cells of the human testis and in some specific cell types from human testicular tumours

The intermediate filament protein nestin is predominantly expressed in some stem/progenitor cells and appears to be a useful molecular tool to characterise tumours originating from precursor cells of neuroectodermal and mesenchymal lineages. Leydig cells originate in the adult testis by differentiation from stem cells and express a variety of neural and neuroendocrine markers. The possible expression of the neural stem cell marker nestin in Leydig cells and testicular tumour cells was determined by analysing the patterns of nestin expression in normal and pathological human testes by Western blot and immunohistochemical methods. In normal testis, nestin was found in some vascular endothelial cells, a subset of peritubular spindle-shaped cells and some Leydig cells; spermatogenic and Sertoli cells were unstained. In normal Leydig cells, nestin was distributed in the perinuclear cytoplasm and accumulated in the crystalloids of Reinke with ageing. In non-tumour pathologies (cryptorchidism, impaired spermatogenesis), the seminiferous tubules were immunonegative, whereas hyperplastic Leydig cells showed cytoplasmic immunolabelling. In testicular malignancies, nestin was localised in the Sertoli cells of the seminiferous tubules affected with intratubular germ cell neoplasia, in the hyperplastic Leydig cells associated with these tumours and in some components (mesenchymal and neuroepithelial cells) of teratomas; spermatocytic and non-spermatocytic seminomas were unstained. Some vascular endothelial cells were immunolabelled in all tumour samples. Thus, nestin is expressed in a population of normal and hyperplastic Leydig cells and in Sertoli cells in the presence of intratubular germ-cell neoplasia. Nestin may be a good marker for identifying components of testicular teratomas.The two first authors participated equally in this workThis work was supported by a grant from the Fondo de Investigaciones Sanitarias (FIS 02/3003 to M.V.T. Lobo)     

Tumour-associated immunoglobulins and host cell infiltration

Summary Using a variety of techniques, we have investigated the possible interrelationship between tumour-associated immunoglobulins and infiltrating host cells. Initial cytological and histological studies revealed that tumour models which normally have a high immunoglobulin content also contained a relatively high proportion of host cells which could bind immunoglobulin by cytophilic processes. Rosette assays on tumour cell suspensions from mice also challenged with SRBC suggested that some of the immunoglobulin present may be non-specifically bound to host cells or synthesized in situ. Evidence that the local production of immunoglobulin could occur was obtained by protein A-PFC analysis of tumour cell suspensions. These studies also revealed that there were many more IgA-secreting cells within tumours than in spleen or lymph nodes. Finally, immunoglobulin analysis on gradient-separated tumour cell suspensions indicated that immunoglobulin was associated with both tumour and host cell-enriched (i.e., Fc-positive) fractions. These observations emphasize that much of the immunoglobulin previously noted in tumour eluates may not be associated with tumour cells as such, but may be bound to or synthesized by infiltrating host lymphoreticular cells.     

Stage-specific inductive signals in the Drosophila neuroectoderm control the temporal sequence of neuroblast specification

One of the initial steps of neurogenesis in the Drosophila embryo is the delamination of a stereotype set of neural progenitor cells (neuroblasts) from the neuroectoderm. The time window of neuroblast segregation has been divided into five successive waves (S1-S5) in which subsets of neuroblasts with specific identities are formed. To test when identity specification of the various neuroblasts takes place and whether extrinsic signals are involved, we have performed heterochronic transplantation experiments. Single neuroectodermal cells from stage 10 donor embryos (after S2) were transplanted into the neuroectoderm of host embryos at stage 7 (before S1) and vice versa. The fate of these cells was uncovered by their lineages at stage 16/17. Transplanted cells adjusted their fate to the new temporal situation. Late neuroectodermal cells were able to take over the fate of early (S1/S2) neuroblasts. The early neuroectodermal cells preferentially generated late (S4/S5) neuroblasts, despite their reduced time of exposure to the neuroectoderm. Furthermore, neuroblast fates are independent from divisions of neuroectodermal progenitor cells. We conclude from these experiments that neuroblast specification occurs sequentially under the control of non-cell-autonomous and stage-specific inductive signals that act in the neuroectoderm.     

NUCLEAR PROTEIN PATTERNS IN DEVELOPING AND ADULT BRAIN AND IN ETHYLNITROSOUREA-INDUCED NEUROECTODERMAL TUMOURS OF THE RAT1,2

In a comparative study, the patterns of histones and non-histone proteins were analysed in the chromatin of foetal (18th day of gestation), 10-day-old, and adult BD IX-rat brain, as well as in the chromatin of two ethylnitrosourea-induced neuroectodermal tumours (TV1A1 and GV1A1) and the corresponding malignant cell culture lines TV1C1 and GV1C1. Separation of nuclear proteins at high resolution was obtained by electrophoresis in 15% and 10% polyacrylamide gels containing urea (2·5 m or 6·25 m ). In spite of an overall similarity, significant quantitative and qualitative differences were observed between the respective non-histone proteins banding patterns of normal brain and the neoplastic cells analysed. The non-histone protein banding patterns of brain (∼40 different bands) at different stages of development revealed both quantitative differences and the presence of particular bands characteristic of foetal or adult brain, respectively. Both the‘foetal’and‘adult’non-histone protein bands also appeared in the electrophoretograms of the neoplastic neuroectodermal cells.     

Causes and consequences of tumour acidity and implications for treatment

Tumour cells have a lower extracellular pH (pHe) than normal cells; this is an intrinsic feature of the tumour phenotype, caused by alterations either in acid export from the tumour cells or in clearance of extracellular acid. Low pHe benefits tumour cells because it promotes invasiveness, whereas a high intracellular pH (pHi) gives them a competitive advantage over normal cells for growth. Molecular genetic approaches have revealed hypoxia-induced coordinated upregulation of glycolysis, a potentially important mechanism for establishing the metabolic phenotype of tumours. Understanding tumour acidity opens up new opportunities for therapy.     

The parathyroid hormone-related protein associated with malignancy is secreted by neuroendocrine tumors

We have demonstrated the production of the PTH-related protein (PTHrP) associated with hypercalcemia of malignancy by human neuroendocrine cell lines that also produce calcitonin gene products and chromogranin A. PTHrP was demonstrable in the cells by immunocytochemistry and immunoassay and Northern analysis of the cells revealed the presence of multiple mRNAs for PTHrP. The cell lines also secreted PTHrP in a regulated fashion, with the most potent secretagogue being phorbol. Thus, PTHrP is secreted by neuroendocrine cells and it may have neuroectodermal lineage. The coexpression of calcitonin gene products and chromogranin A, also neuroendocrine, with PTHRP may influence its secretion and ultimate biological effects in vivo.     

Analysis of pp60c-src protein kinase activity in human tumor cell lines and tissues

We have evaluated the level of pp60c-src protein kinase activity in a variety of human tumor tissues and human tumor cell lines, and have estimated the abundance of the c-src protein in several of these tissues and cell lines. All cell lines derived from tumors of neuroectodermal origin that express a neural phenotype were found to possess c-src molecules with high levels of tyrosine-specific protein kinase activity. In contrast, cell lines derived from tumors of neuroectodermal origin that do not express neural characteristics, such as glioblastomas and melanomas, were found to have pp60c-src molecules with low levels of protein kinase activity. A similar pattern was observed when we analyzed the activity of c-src molecules extracted directly from corresponding tumor tissues. Analysis of human tumor cell lines derived from tissues other than those of neuroectodermal origin revealed that pp60c-src protein kinase activity was low in most cases. Exceptions to this observation were all rhabdomyosarcoma, osteogenic sarcoma, Ewing's sarcoma, and colon carcinoma lines tested. Comparison of pp60c-src kinase activity in normal skeletal muscle and rhabdomyosarcoma tissue and in normal breast tissue and breast adenocarcinoma tissue revealed that pp60c-src kinase activity was specifically elevated in the tumor tissues in both cases. However, the amount of pp60c-src protein in both normal and tumor tissues was found to be similar. These observations suggest that increases in the specific activity of the pp60c-src phosphotransferase in some rhabdomyosarcomas and breast carcinomas may be a characteristic acquired during the malignant transformation of the cells that is retained in cell lines established from these tumors.     

Fine needle aspiration (FNA)-induced necrosis of tumours of the thyroid

Two cases of necrosis of thyroid oxyphilic tumours following FNA are reported. the first patient received surgery 4 weeks after FNA and histological examination revealed an encapsulated and totally necrotic tumour 2 cm in diameter. In the second patient surgery was performed after 25 days. Histological examination showed a 0.7 cm diameter tumour consisting mainly of fibrous tissue with residual oxyphilic tumour cells only in a small peripheral rim. In both patients no capsular or vascular invasion and no blood vessel thrombosis were present. A review of the literature revealed that oxyphilic tumours are susceptible to post-FNA necrosis, which might be due to the compromised vascular supply after FNA in conjunction with the intrinsic energy deficiency of oncocytic tumour cells.     

Misexpression of mouse porcupine isoforms modulates the differentiation of P19 embryonic carcinoma cells

Drosophila porcupine (porc) encodes an ER membrane protein that is required for the processing of the Drosophila Wnt family. Homologs of porc have been identified in various multicellular organisms and have been implicated in the biosynthesis of Wnt proteins. In contrast to Drosophila, vertebrates generate four different porc mRNAs (A-D) by alternative splicing. Murine porcD (MporcD) mRNA levels transiently increase during the neuroectodermal differentiation of P19 cells, but diminish during mesodermal differentiation. P19 cells constitutively expressing mouse porcA (MporcA), but not MporcD, undergo apoptosis by the induction of neuroectodermal differentiation. Meanwhile, P19 cells constitutively expressing MporcD, but not MporcA, do not adopt mesodermal cell morphology and fail to express myf-5 when induced to mesodermal differentiation. These results therefore demonstrate that the alternative splicing of Mporc is regulated in a cell-type specific manner, and the resulting Mporc isoforms have different functions in the neuroectodermal and mesodermal differentiation of P19 cells.